CN108743622A - Immunity of organisms and muscle power are improved using bear gall powder and alleviate the method for organism fatigue - Google Patents
Immunity of organisms and muscle power are improved using bear gall powder and alleviate the method for organism fatigue Download PDFInfo
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- CN108743622A CN108743622A CN201810773488.XA CN201810773488A CN108743622A CN 108743622 A CN108743622 A CN 108743622A CN 201810773488 A CN201810773488 A CN 201810773488A CN 108743622 A CN108743622 A CN 108743622A
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- 239000008223 sterile water Substances 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 229960005202 streptokinase Drugs 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 235000019640 taste Nutrition 0.000 description 1
- 230000001732 thrombotic effect Effects 0.000 description 1
- 210000001541 thymus gland Anatomy 0.000 description 1
- 239000004408 titanium dioxide Substances 0.000 description 1
- 210000003371 toe Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
- 150000003626 triacylglycerols Chemical class 0.000 description 1
- 230000005909 tumor killing Effects 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 210000003556 vascular endothelial cell Anatomy 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 239000000052 vinegar Substances 0.000 description 1
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- 235000020234 walnut Nutrition 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 210000002417 xiphoid bone Anatomy 0.000 description 1
- 239000009131 zhiling Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/37—Digestive system
- A61K35/413—Gall bladder; Bile
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
Immunity of organisms and muscle power are improved the invention discloses application bear gall powder and alleviate the method for organism fatigue.Bear gall powder can be used for adjusting blood fat, reduce total cholesterol, prevent cardiovascular and cerebrovascular disease, prevention of arterial atherosis;Bear gall powder obtained by the preparation method and this method of the bear gall powder.The present invention makes material ice crystalization then by freeze-drying during preparing bear gall powder, and thus gained freeze-drying bear gall powder can be referred to as ice crystal state bear gall powder in the present invention, or can be described as ice crystal bear gall powder.Especially the present invention can be referred to as the bear gall powder of gloomy precious bear gall powder without any auxiliary material and chemical addition agent.Also particularly find that ice crystal state bear gall powder of the present invention is not only able to more effectively adjust blood fat, reduces total cholesterol, it can also be more efficiently used for preventing cardiovascular and cerebrovascular disease, prevention of arterial atherosis, entity tumor and hematological system tumor are prevented and treated, can more particularly be efficiently used for improving immunity of organisms and muscle power alleviates organism fatigue.
Description
Technical field
The invention belongs to the technical fields of medical and health product, are related to a kind of bear gall powder being prepared by special process,
Further relate to the preparation method of this bear gall powder.The present invention in the preparation process of bear gall powder by making material ice crystalization then pass through
Freeze-drying, bear gall powder obtained are presented excellent technique effect, excellent skill are especially for example presented in physics and chemistry properties
Art effect.Particularly, the present invention is diluted with a small amount of water in advance when preparing bear gall powder, and progress depth is cold after being filled with two kinds of gases
Freeze, so that contained active constituent in bear gall juice is in a kind of ice crystal state, then distil, thus gained is freeze-dried bear gall powder
It can be referred to as ice crystal state bear gall powder in the present invention, either can be described as ice crystal bear gall powder or can be described as gloomy precious bear gall powder.It is special
The bear gall powder of gloomy precious bear gall powder of new generation can be referred to as without any auxiliary material and chemical addition agent by not being the present invention.The present invention
Also particularly find that ice crystal state bear gall powder of the present invention is not only able to more effectively adjust blood fat, reduces total cholesterol, moreover it is possible to more have
Effect ground is for preventing the special efficacies such as cardiovascular and cerebrovascular disease, prevention of arterial atherosis.More particularly, ice crystal state of the present invention
Bear gall powder can also be used to prevent or treat gall stone disease and improve gallbladder function, in addition, the bear gall of ice crystal state of the present invention
Powder can also be used to prevent or treat hepatopathy and liver fibrosis and improve liver function;In addition, the bear gall of ice crystal state of the present invention
Powder can also be used to prevent or treat entity tumor and hematological system tumor;In addition, the bear gall powder of ice crystal state of the present invention can also
For inhibition thrombosis, inhibit platelet aggregation, prevention cerebral ischemia and ischemic cerebral disease.In addition, the bear of ice crystal state of the present invention
Courage powder can also be used to improve immunity of organisms, improve muscle power, alleviate organism fatigue.
Background technology
More editions《Chinese Pharmacopoeia》The medicinal material of the bile with bear gall and/or its secretion is recorded or comprising its preparation.Example
Such as, version in 2005《Chinese Pharmacopoeia》It is ursidae animal black bear Selenarctos that medicinal material bear gall has been recorded in one annex page 24
The drying gall-bladder of thibetanus Cuvier or brown bear Ursus arctos Linnaeus.Version in 2010《Chinese Pharmacopoeia》One
It is that ursidae animal black bear Selenarctos thibetanus Cuvier draw through operation on gallbladder that bear gall powder has been recorded in annex page 27
Flow dry product obtained by bile;This edition pharmacopeia one has also recorded bear gall capsule, bear gall pills for curing heart disease/pellet, bear gall hemorrhoid spirit cream etc.
It is preparation made of raw material including bear gall powder.Version in 2015《Chinese Pharmacopoeia》One recorded bear gall capsule, bear gall pills for curing heart disease,
Bear gall hemorrhoid spirit cream, bear gall ZHILING SHUAN etc. are preparation made of raw material including bear gall powder.
Currently, substitute of the dry product of drain-out bear gall's bile as natural bear gall is used as medicine and examines listing, life
Entitled bear gall powder is a kind new medicine of health ministry approval.Bear gall powder is that ursidae animal black bear drains courage by operation on gallbladder
Dry product obtained by juice, cold in nature, bitter enter liver, courage, spleen, stomach, large intestine channel, have heat-clearing, soothing the liver, the work(of improving eyesight.Modern study
It was found that bear gall powder chemical composition is more complicated, main ursodesoxycholic acid containing mating type (UDCA), chenodeoxycholic acid (CDCA), courage
Sour (CA), deoxycholic aicd (DCA), Tauro ursodesoxy cholic acid (TUDCA), Taurochenodeoxycholic Acid (TCDCA) and cholesterol, courage
Pigment, amino acids, protein, peptide, aliphatic acid, trace element etc..Wherein ursodesoxycholic acid is important characteristic component.Mesh
It is preceding to be clinically mainly used in the diseases in the liver and gallbladder such as gall stone, fatty liver, cholecystitis, virus hepatitis, chronic hepatitis B, eyelid
Herpes zoster, the diseases such as hemorrhoid.
CN103520210A (201310523280.X, Sheng Yuantang) discloses a kind of bear gall powder purification process, including as follows
The step of sequence carries out:(1) polyvinylpyrrolidone (PVP) and sterile water distilled water are added in freshly extd bear gall juice,
Mixing is shaken, clarification is stood, obtains supernatant;(2) absolute ethyl alcohol of 2 times of volumes is added in gained supernatant in step (1),
Bear gall alcohol extract is made;(3) macro porous silica gel pillar is added in bear gall purification obtained by step (2), it is anhydrous using 2 times of volumes 75%
Ethyl alcohol, which divides 3 times, washs silicon gel column, collects cleaning solution;(4) with sterile distilled water to gained cleaning solution in step (3) into
Row dilution so that a concentration of the 16.5% of absolute ethyl alcohol;(5) by bear gall of the gained containing 16.5% absolute ethyl alcohol in step (4)
Juice supernatant is added in cellulose powder CF11 pillars, vortex oscillation mixing, progress bear gall juice separation, under the conditions of 5000rpm, from
After centrifugation, isometric sterile distilled water is added in heart 5min in cellulose powder CF11 pillars, under the conditions of 5000rpm, from
Heart 5min;(6) gained supernatant in collection step (5), is added the 3mol/L NaAc of 1/10 volume and isometric isopropanol,
Mixing, 20 DEG C precipitate, and after the completion of precipitation, under the conditions of 4 DEG C, 12000rpm centrifuges 30min, abandons supernatant, collects precipitation;(7) it is added
75% absolute ethyl alcohol washs the precipitation collected in step (6), after washing, under 4 DEG C of environment, and 12000rpm centrifugations
30min abandons supernatant, collects precipitation;(8) it by gained precipitation in step (7), is dissolved in distilled water, stirs 1 to 3 hour, bear is made
Courage solution, it is 2 to 8 to adjust pH, is heated to 50 DEG C and keeps the temperature, addition compound protease, after enzymolysis, enzyme deactivation, into
Row ultrafiltration membrance filter can be obtained bear gall powder finished product after the supernatant of acquisition is spray-dried.It is believed that bear obtained by the inventive technique
Courage powder, effective content is high, and pigment content is low, no fishy smell, and medical value is promoted.
CN105147729A (201510650516.5, pasture river) discloses a kind of preparation method of bear gall powder, and step includes:
(A), it filters:Fresh drained bear bile is taken, filters, obtains filtrate;(B), it sterilizes:High concentration ethanol, ethyl alcohol are added in filtrate
Ethanol content is 75%~85% in liquid after addition, is then stirred, and is stood, and mixing time is 20~40 minutes, is stood
Time is 24~72 hours;(C), it concentrates:It is concentrated under reduced pressure and removes ethyl alcohol, reduced pressure carries out at 35~45 DEG C, is concentrated under reduced pressure
After obtain sterilizing bear bile fluid;(D), it is freeze-dried:Sterilizing bear bile fluid is freeze-dried, and the method for the freeze-drying is:
Sterilizing bear bile fluid is cooled to 35~40 DEG C, 0.5~3.5h of constant temperature with the rate of temperature fall of 1~2 DEG C/min, it is evacuated to 1~
15Pa, then room temperature is warming up to the heating rate of 1~3 DEG C/min.It is believed that the characteristics of invention utilizes new fresh bear bile to be liquid,
It is miscible with bile using ethyl alcohol, using ethyl alcohol to virus, pathogenic bacteria killing effect, by control ethyl alcohol in liquid content and
Time does not destroy the active constituent in bear gall juice, kills various pathogenic bacteria and virus, ensure clinical application safety, effectively
Property, the defect of traditionally bear gall powder production is overcome, gained bear gall powder appearance is golden yellow, has transparent gloss, greatlys improve
Bear gall powder quality.
The method that CN106386659A (201610755739.2, day is helped) discloses bear gall powder factorial praluction, this method
Selection including black bear breeding raises, takes courage, feeding Chinese herbal medicine, cryogenic temperature freezing drying, specific as follows:(1) black bear breeding
Selection:Select healthy and strong, varietal characteristic is apparent, feeding power is strong, production performance is good, constitution is solid, limb hoof is healthy and strong, 3 years old or more
Black bear do culture bear;(2) it raises:Culture bear is fed using mixed feed, the mixed feed matching in mass ratio
Than for:50~60 parts of corn, 3~5 parts of fish meal, 10~15 parts of barley, 10~15 parts of the soybean fried, 3~5 parts of meat, silkworm chrysalis 3
~5 parts;Nursing starts to take courage when reaching 90 ㎏ or more to black bear weight;(3) courage is taken:Using the painless drainage of the self tubing of animal
Method adopts courage, and specific practice is, between the stomach wall and gall-bladder of black bear, is woven in stomach wall end using the group of black bear itself and manufactures annular
Sphincter, when sphincters contract, can close tube chamber, and when diastole makes tube chamber open, domination or hormone of the sphincter by autonomic nerve
It adjusts, using this characteristic, nutrient solution is fed to black bear when adopting courage, black bear sphincter relaxation, tube chamber is made to open, be disappeared with cotton ball soaked in alcohol
Stainless steel hollow probe after disinfection is probeed into tube chamber channel by the open tube chamber passway of poison, bile i.e. along hollow probe from
Dynamic flow to connects in the cup of bile, takes extraction probe after the completion of courage;(4) Chinese herbal medicine is fed:After the completion of taking courage, fed to black bear
The Chinese herbal medicine of syrup water is added in clothes, continuously feeds 10 days;The Chinese herbal medicine formula is:20~30 parts of Cortex Magnoliae Officinalis, 20~30 parts of green peel,
20~30 parts of Chinese cassia tree, 20~30 parts of spinach;20~30 parts of the fleece-flower root, 20~30 parts of Radix Glycyrrhizae, 20~30 parts of folium isatidis, Radix Isatidis 20
~30 parts;20~30 parts of corn protein powder, 20~30 parts of analysis for soybean powder, 20~30 parts of walnut dregs, 20~30 parts of peanut cypress;The fruit of glossy privet
40~50 parts, 40~50 parts of adlay, 5~10 parts of dried orange peel, 5~10 parts of ginger;Said herbal medicine is crushed and is added after mixing
Enter syrup water and feeds black bear;(5) cryogenic temperature freezing drying:By the bear gall juice of taking-up in advance -60 DEG C of temperature and it is following freeze, so
It is placed under vacuum state, the water sublimed contained obtains dry bear gall powder.It is believed that the inventive method was prepared
Bear gall powder quality is good, yield is high and is not injured to black bear body.
CN102114044A (201010617773.6, Gui Zhentang) discloses a kind of extracting method of bear gall juice, in bear
Wound is cut on gall-bladder, and forms fistula with musculature, is tightened;Fistula mouth acquisition bile is squeezed after recovery from illness.This method can be used for
It is discontinuous, at regular time and quantity and long-term acquisition, and bear itself will not be impacted using bile.The invention discloses a kind of excellent
Matter bear gall powder and preparation method thereof, the bear gall powder are in glassy yellow fluorescence under 365nm ultraviolet lamps, there is 3 chromatographies at 198nm
Peak;Preparation method include by bile stand, separation, crushed after being dried sieving to get.It is believed that bear gall powder appearance prepared by the invention
Pure color, Tauro ursodesoxy cholic acid content is high in bear gall powder, nonhygroscopic, is conducive to storage and transportation and processing.
CN103040869A (201310027917.6, Kang Ao) discloses a kind of artificial bear gall powder, and it is original that it, which is by fowl courage,
Material is added cow-bezoar sodium ursodexoxycholate and is prepared, and wherein cow-bezoar sodium ursodexoxycholate and the weight proportion of fowl courage is:Cow-bezoar bear
20-40 parts of deoxysodium cholate, 80-60 parts of fowl courage.The present invention also provides the preparation methods of the artificial bear gall powder.Invention is not only matched
Fang Kexue is unique, raw material is easy to get, is simple for process, is of low cost, ensures quality, ensure curative effect, save the wild animals, and inherent
Quality and appearance character are all almost close with natural bear gall, provide a simplicity, economy, safe and environment-friendly, and quality and appearance are all
It is the ideal substitute of natural bear gall with the extremely similar artificial bear gall powder of natural bear gall.
CN1311002A (00102073.0, Shi Lixia) discloses a kind of refined bear gall powder and preparation method thereof, belongs to pharmacy
The common bear gall powder in field easy moisture absorption caking, there is that tooth gluing sense, taste are extremely stench, extremely bitter, impurity is more, absorption is slow, and property is unstable easy
Corruption, effective component content is low, limits the application range of bear gall powder, and when oral application is commonly incorporated into flavoring agent, mitigates bad smell.
The invention is refined using ethanol extraction method, ethyl alcohol extraction activated carbon decolorizing method and ethyl acetate partition method purification bear gall powder
Rear bear gall pink colour it is shallow it is fine and smooth, without tooth gluing sense, without bad smell, bitter taste is light, be not easy corruption, be not easy moisture absorption caking, property is stablized, have
Effect component content is averagely increased to 2 times or more.Extend period of storage, expands application range, improve curative effect of medication.
CN106038601A (201610368003.X, great Yi) discloses a kind of high-content, high-purity, low fishy smell bear gall powder
Preparation method, specific method is to be sufficiently mixed bear gall powder, activated carbon and filter aid, then carries mixture with ethanol solution
It is feminine gender to take into extracting solution cholanic acid, collects ethanol extract, concentrates and recycle ethyl alcohol, collects concentrate, dry and obtain;Institute
It is water and the insoluble silicate mineral of alcohol to state filter aid.It is believed that the method for the invention is simple, more operability, controllability
Good, content of the bear gall powder obtained in terms of Tauro ursodesoxy cholic acid reaches as high as 45% or more, the rate of recovery nearly 100%, with weight
The rate of recovery is calculated up to 85% or more, this method is suitable for industrialization large-scale production, at low cost, can be used in preparing special system
High-quality high-content, high-purity, low fishy smell bear gall powder needed for agent enterprise.
However, this field still expect to have new method preparing with the bear gall powder of advantageous property and its medical treatment and
The purposes of health care, such as adjust blood fat, reduce the purposes such as total cholesterol, anti-cardiovascular and cerebrovascular disease, prevention of arterial atherosis, with
And the purposes of the medical and health care described in denomination of invention for example of the invention.
Invention content
The purpose of the present invention is to provide a kind of new methods to prepare the bear gall powder with advantageous property.The present invention is another
It is designed to provide a kind of bear gall powder with advantageous property.In addition, the present invention also provides the bear gall powders of the invention prepared
Using;The especially purposes of its medical and health care, such as adjust blood fat, reduce total cholesterol, anti-cardiovascular and cerebrovascular disease, prevent to move
The purposes such as pulse atherosclerosis, and medical and health care described in denomination of invention for example of the invention purposes.
For this purpose, first aspect present invention provides a kind of bear gall powder, it is the burnished gold or foresythia of hyaloid gloss
Powder, shows glassy yellow fluorescence at 365nm, and for example≤1%, and its solubility in water is more than 10g/ to hydroscopicity≤2%
100ml, the solubility in 90% ethyl alcohol are more than 1.2g/100ml, and the pH value of 5% aqueous solution is in 6.0~7.5 ranges.
Bear gall powder according to a first aspect of the present invention, the solubility in water are 13.6~14.2g/100ml.
Bear gall powder according to a first aspect of the present invention, the solubility in 90% ethyl alcohol are 1.25~2g/100ml.
Bear gall powder according to a first aspect of the present invention, the pH value of 5% aqueous solution is in 6.0~7.0 ranges.
Bear gall powder according to a first aspect of the present invention is prepared by a method comprising the following steps to obtain:
(1) the bear gall juice by the acquisition of no Tube Drain method filters, and obtains filtrate, is diluted with water, obtain bear bile fluid, in 2~6 DEG C of temperature
12 hours are stood at degree, then carries out subsequent operation;
(2) it is filled with 20~30min of nitrogen into bear bile fluid obtained by step (1), is then filled with 10~20min of carbon dioxide;
(3) bear bile fluid handled through aforesaid operations is inclined in material disc, be immediately placed in advance be refrigerated to -45 DEG C it is below
In freeze drier, freeze 2~5 hours;Then -25~-20 DEG C are warming up to 0.25~0.35 DEG C of speed per minute, kept
1~1.5 hour;Then it is warming up to -5~0 DEG C with 0.25~0.35 DEG C of speed per minute, is kept for 2~3 hours;Then with every
The speed of 0.25~0.35 DEG C of minute is warming up to 10~15 DEG C, is kept for 4~5 hours;Then with 0.25~0.35 DEG C per minute
Speed is warming up to 30~35 DEG C, is kept for 2~3 hours, terminates freeze-drying program;
(4) by the dried lump in material disc be ground into can by the fine powder that 50 mesh sieve to get.
In bear gall powder according to a first aspect of the present invention, wherein step (1) plus the water of 0.5~1 times of amount dilutes.
When inflatable body filled in a manner of bubbling in bear gall powder according to a first aspect of the present invention, wherein step (2)
Gas.
Bear gall powder according to a first aspect of the present invention, wherein step are filled with 25~30min of nitrogen in (2).
Bear gall powder according to a first aspect of the present invention, wherein step are filled with 15~20min of carbon dioxide in (2).
Bear bile fluid is inclined in material disc in bear gall powder according to a first aspect of the present invention, wherein step (3), and liquid layer depth is small
In 5cm.
Bear bile fluid is inclined in material disc in bear gall powder according to a first aspect of the present invention, wherein step (3), and liquid layer depth is small
In 2.5cm.
Bear bile fluid is inclined in material disc in bear gall powder according to a first aspect of the present invention, wherein step (3), and liquid layer depth is
1.5~2.5cm.
In bear gall powder according to a first aspect of the present invention, wherein step (3), make material disc be immediately placed on be refrigerated in advance-
In 55~-45 DEG C of freeze drier.
In bear gall powder according to a first aspect of the present invention, wherein step (3), make material disc be immediately placed on be refrigerated in advance-
In 50~-45 DEG C of freeze drier.
In bear gall powder according to a first aspect of the present invention, wherein step (3), make material disc be immediately placed on be refrigerated in advance-
In 50~-45 DEG C of freeze drier, freeze 3~4 hours;Then -25~-20 DEG C are warming up to 0.3 DEG C of speed per minute,
It is kept for 1~1.5 hour;Then it is warming up to -5~0 DEG C with 0.3 DEG C of speed per minute, is kept for 2~3 hours;Then with per minute
0.3 DEG C of speed is warming up to 10~15 DEG C, is kept for 4~5 hours;Then 30~35 DEG C are warming up to 0.3 DEG C of speed per minute,
It is kept for 2~3 hours, terminates freeze-drying program.
Bear gall powder according to a first aspect of the present invention, moisture are 0.4~1%.
It has been unexpectedly discovered that bear bile fluid is made to be filled with nitrogen and carbon dioxide successively in advance, it is not only able to significantly increase
Add the solubility of gained bear gall powder, and active constituent Tauro ursodesoxy cholic acid (C26H45NO6S) display in gained bear gall powder
Go out more excellent stability, this is the prior art can not predict at all.
In addition, the present invention is diluted with a small amount of water in advance when preparing bear gall powder, cryogenic refrigeration is carried out after being filled with two kinds of gases,
So that contained active constituent in bear gall juice is in a kind of ice crystal state, then distil, thus gained freeze-drying bear gall powder exists
It can be referred to as ice crystal state bear gall powder in the present invention.
Further, second aspect of the present invention provides the method for preparing bear gall powder, includes the following steps:
(1) the bear gall juice by the acquisition of no Tube Drain method filters, and obtains filtrate, is diluted with water, obtain bear bile fluid, in 2~6 DEG C of temperature
12 hours are stood at degree, then carries out subsequent operation;
(2) it is filled with 20~30min of nitrogen into bear bile fluid obtained by step (1), is then filled with 10~20min of carbon dioxide;
(3) bear bile fluid handled through aforesaid operations is inclined in material disc, be immediately placed in advance be refrigerated to -45 DEG C it is below
In freeze drier, freeze 2~5 hours;Then -25~-20 DEG C are warming up to 0.25~0.35 DEG C of speed per minute, kept
1~1.5 hour;Then it is warming up to -5~0 DEG C with 0.25~0.35 DEG C of speed per minute, is kept for 2~3 hours;Then with every
The speed of 0.25~0.35 DEG C of minute is warming up to 10~15 DEG C, is kept for 4~5 hours;Then with 0.25~0.35 DEG C per minute
Speed is warming up to 30~35 DEG C, is kept for 2~3 hours, terminates freeze-drying program;
(4) by the dried lump in material disc be ground into can by the fine powder that 50 mesh sieve to get.
In method according to a second aspect of the present invention, wherein step (1) plus the water of 0.5~1 times of amount dilutes.
When inflatable body it is inflated in a manner of bubbling in method according to a second aspect of the present invention, wherein step (2).
Method according to a second aspect of the present invention, wherein step are filled with 25~30min of nitrogen in (2).
Method according to a second aspect of the present invention, wherein step are filled with 15~20min of carbon dioxide in (2).
Bear bile fluid is inclined in material disc in method according to a second aspect of the present invention, wherein step (3), and liquid layer depth is less than
5cm。
Bear bile fluid is inclined in material disc in method according to a second aspect of the present invention, wherein step (3), and liquid layer depth is less than
2.5cm。
Bear bile fluid is inclined in material disc in method according to a second aspect of the present invention, wherein step (3), and liquid layer depth is
1.5~2.5cm.
In method according to a second aspect of the present invention, wherein step (3), so that material disc is immediately placed on and be refrigerated to -55 in advance
In~-45 DEG C of freeze drier.
In method according to a second aspect of the present invention, wherein step (3), so that material disc is immediately placed on and be refrigerated to -50 in advance
In~-45 DEG C of freeze drier.
In method according to a second aspect of the present invention, wherein step (3), so that material disc is immediately placed on and be refrigerated to -50 in advance
In~-45 DEG C of freeze drier, freeze 3~4 hours;Then -25~-20 DEG C are warming up to 0.3 DEG C of speed per minute, protected
It holds 1~1.5 hour;Then it is warming up to -5~0 DEG C with 0.3 DEG C of speed per minute, is kept for 2~3 hours;Then with per minute
0.3 DEG C of speed is warming up to 10~15 DEG C, is kept for 4~5 hours;Then 30~35 DEG C are warming up to 0.3 DEG C of speed per minute,
It is kept for 2~3 hours, terminates freeze-drying program.
Method according to a second aspect of the present invention, obtained by bear gall powder be hyaloid gloss burnished gold or cadmium yellow
Color powder, shows glassy yellow fluorescence at 365nm, and for example≤1%, and its solubility in water is more than hydroscopicity≤2%
10g/100ml, the solubility in 90% ethyl alcohol are more than 1.2g/100ml, and the pH value of 5% aqueous solution is in 6.0~7.5 ranges
It is interior.
Method according to a second aspect of the present invention, obtained by bear gall powder solubility in water be 13.6~14.2g/
100ml.In another embodiment, the moisture of bear gall powder of the present invention is less than 3%.
Method according to a second aspect of the present invention, obtained by solubility of the bear gall powder in 90% ethyl alcohol be 1.25~
2g/100ml。
Method according to a second aspect of the present invention, obtained by bear gall powder its 5% aqueous solution pH value 6.0~7.0
In range.
Method according to a second aspect of the present invention, obtained by bear gall powder moisture be 0.4~1%.
Further, third aspect present invention provides bear gall powder and is preparing for adjusting blood fat, reducing total cholesterol, pre-
Anti- cardiovascular and cerebrovascular disease, prevention of arterial atherosis product in purposes;And it is swollen for preventing or treating entity preparing
Purposes in the product of tumor and hematological system tumor;And it is preparing for preventing or treating entity tumor and hematological system tumor
Product in purposes;And it is preparing for inhibition thrombosis and platelet aggregation and is preventing cerebral ischemia and ischemic cerebral disease
Product in purposes.And it is preparing for the use in improving immunity of organisms, improving product that is physical, alleviating organism fatigue
On the way.
Purposes according to a third aspect of the present invention, wherein the entity tumor and hematological system tumor are selected from:Lung cancer, white blood
Disease, sarcoma and liver cancer.
Purposes according to a third aspect of the present invention, wherein the product is drug, health food.
Purposes according to a third aspect of the present invention, wherein the bear gall powder is the burnished gold or foresythia of hyaloid gloss
Powder, shows glassy yellow fluorescence at 365nm, and for example≤1%, and its solubility in water is more than 10g/ to hydroscopicity≤2%
100ml, the solubility in 90% ethyl alcohol are more than 1.2g/100ml, and the pH value of 5% aqueous solution is in 6.0~7.5 ranges.
Purposes according to a third aspect of the present invention, wherein the solubility of the bear gall powder in water is 13.6~14.2g/
100ml。
Purposes according to a third aspect of the present invention, wherein solubility of the bear gall powder in 90% ethyl alcohol be 1.25~
2g/100ml。
Purposes according to a third aspect of the present invention, wherein the pH value of 5% aqueous solution of the bear gall powder is in 6.0~7.0 ranges
It is interior.
Purposes according to a third aspect of the present invention, wherein the bear gall powder is to be prepared by a method comprising the following steps
It arrives:
(1) the bear gall juice by the acquisition of no Tube Drain method filters, and obtains filtrate, is diluted with water, obtain bear bile fluid, in 2~6 DEG C of temperature
12 hours are stood at degree, then carries out subsequent operation;
(2) it is filled with 20~30min of nitrogen into bear bile fluid obtained by step (1), is then filled with 10~20min of carbon dioxide;
(3) bear bile fluid handled through aforesaid operations is inclined in material disc, be immediately placed in advance be refrigerated to -45 DEG C it is below
In freeze drier, freeze 2~5 hours;Then -25~-20 DEG C are warming up to 0.25~0.35 DEG C of speed per minute, kept
1~1.5 hour;Then it is warming up to -5~0 DEG C with 0.25~0.35 DEG C of speed per minute, is kept for 2~3 hours;Then with every
The speed of 0.25~0.35 DEG C of minute is warming up to 10~15 DEG C, is kept for 4~5 hours;Then with 0.25~0.35 DEG C per minute
Speed is warming up to 30~35 DEG C, is kept for 2~3 hours, terminates freeze-drying program;
(4) by the dried lump in material disc be ground into can by the fine powder that 50 mesh sieve to get.
In purposes according to a third aspect of the present invention, wherein step (1) plus the water of 0.5~1 times of amount dilutes.
When inflatable body it is inflated in a manner of bubbling in purposes according to a third aspect of the present invention, wherein step (2).
Purposes according to a third aspect of the present invention, wherein step are filled with 25~30min of nitrogen in (2).
Purposes according to a third aspect of the present invention, wherein step are filled with 15~20min of carbon dioxide in (2).
Bear bile fluid is inclined in material disc in purposes according to a third aspect of the present invention, wherein step (3), and liquid layer depth is less than
5cm。
Bear bile fluid is inclined in material disc in purposes according to a third aspect of the present invention, wherein step (3), and liquid layer depth is less than
2.5cm。
Bear bile fluid is inclined in material disc in purposes according to a third aspect of the present invention, wherein step (3), and liquid layer depth is
1.5~2.5cm.
In purposes according to a third aspect of the present invention, wherein step (3), so that material disc is immediately placed on and be refrigerated to -55 in advance
In~-45 DEG C of freeze drier.
In purposes according to a third aspect of the present invention, wherein step (3), so that material disc is immediately placed on and be refrigerated to -50 in advance
In~-45 DEG C of freeze drier.
In purposes according to a third aspect of the present invention, wherein step (3), so that material disc is immediately placed on and be refrigerated to -50 in advance
In~-45 DEG C of freeze drier, freeze 3~4 hours;Then -25~-20 DEG C are warming up to 0.3 DEG C of speed per minute, protected
It holds 1~1.5 hour;Then it is warming up to -5~0 DEG C with 0.3 DEG C of speed per minute, is kept for 2~3 hours;Then with per minute
0.3 DEG C of speed is warming up to 10~15 DEG C, is kept for 4~5 hours;Then 30~35 DEG C are warming up to 0.3 DEG C of speed per minute,
It is kept for 2~3 hours, terminates freeze-drying program.
Purposes according to a third aspect of the present invention, wherein the moisture of the bear gall powder is 0.4~1%.
In the above-mentioned preparation method of the invention the step of, although the specific steps of its description are in certain details or language
Step different from description and described in the preparation example of following detailed description part, however, people in the art
Member can summarize approach described above step completely according to the detailed disclosure of full text of the present invention.
Any embodiment of the either side of the present invention, can be combined with other embodiments, as long as they are not
It will appear contradiction.In addition, in any embodiment of either side of the present invention, any technical characteristic can be adapted for other realities
The technical characteristic in scheme is applied, as long as they are not in contradiction.The invention will be further described below.
All documents recited in the present invention, their full content are incorporated herein by reference, and if these are literary
When offering expressed meaning and the inconsistent present invention, it is subject to the statement of the present invention.In addition, the various terms that use of the present invention and
Phrase has that well known to a person skilled in the art general senses, nonetheless, the present invention remain desirable at this to these terms and
Phrase is described in more detail and explains, the term and phrase referred to is if any inconsistent with common art-recognized meanings, with institute's table of the present invention
Subject to the meaning stated.
Bear gall is ursidae animal black bear (Selenarctos thibetanus Cuvier) or brown bear (Ursus arctos
Linnaeus drying gall-bladder).Substitute of the dry product of drain-out bear gall as natural bear gall is used as medicine and examines listing,
It is named as bear gall powder, is a kind new medicine of health ministry approval.Bear gall powder is ursidae animal black bear (Selenarctos
Thibetanus Cuvier) by dry product obtained by operation on gallbladder bile drainage, cold in nature, bitter enters liver, courage, spleen, stomach, big
Intestines pass through, and have heat-clearing, soothing the liver, the work(of improving eyesight.Modern study finds that bear gall powder chemical composition is more complicated, mainly contains mating type
Ursodesoxycholic acid (UDCA), chenodeoxycholic acid (CDCA), cholic acid (CA), deoxycholic aicd (DCA), Tauro ursodesoxy cholic acid
(TUDCA), Taurochenodeoxycholic Acid (TCDCA) and cholesterol, bilins, amino acids, protein, peptide, aliphatic acid, micro-
Secondary element etc..Wherein ursodesoxycholic acid is important characteristic component.Clinically it is mainly used in gall stone, fatty liver, courage at present
The diseases in the liver and gallbladder such as capsulitis, virus hepatitis, chronic hepatitis B, eyelid herpes zoster, the diseases such as hemorrhoid.
Bear gall powder liver protection pharmacologically, cholagogic, litholytic effect.Have research by Female guinea pigs experiments have shown that bear gall
Powder can reduce cholesterol concentration, improve bile acid concentration, alleviate fatty degeneration of liver caused by high caloric diet high in fat, it was demonstrated that bear
Courage powder, which has, treats and prevents adipohepatic effect.In addition, TUDCA may influence the intestines liver cycle of bile acid, increase total courage
The content of UDCA in juice acid pond, reduces the content to the virose bile acid of liver cell, and prolonged application does not have apparent toxin for liver pair
Effect.There is research to induce N-nitrosodimethylamine (DMN) inhibiting effect of rat liver fibrosis by inquiring into bear gall powder.As a result it demonstrate,proves
Bright, bear gall powder has the function of that DMN is preferably inhibited to induce rat liver fibrosis.Its mechanism of action may be thin with inhibition Ku Pufu
Born of the same parents (Kupffer cell, KC), reduce the secretion of cell factor, inhibit hepatic stellate cells (hepatic satellite cell,
HSC activation) and conversion, synthesis and the secretion for reducing collagenous fibres are related.Have research by the external dissolve stone of bear gall powder, in vivo
The each group of data comparison of anti-stone experiment, it is found that bear gall powder has dissolve stone work to bile pigment calculus, cholesterol stone and mixed type calculus
With.Analysis of experimental results shows that bear gall powder can reduce bile cholesterol content, increases total bile acid content, can be obviously improved
Bile of forming calculus ingredient.Have research by rabbit experimental studies have found that, bear gall powder significantly reduce rabbit bait cholesterol courage
The incidence of calculus reduces the content of free cholesterol in bile, increases the content of total bile acid, it was demonstrated that bear gall powder has prevention
The effect that bait gall stone is formed.Chenodeoxycholic acid, ursodesoxycholic acid can make the reduction of bile cholesterol content, this mainly leads to
Cross inhibit cholesterol small intestine absorption, inhibit the activity of methyl glutaryl coenzyme A reductase to reducing cholesterol biosynthesis and
It realizes.
The antitumaous effect of bear gall powder pharmacologically.Have research by with bear gall to K562, SP20 tumour cell and mouse
S180 ascites carcinoma inhibiting tumor assays find that 1mg/ml concentration bear bile fluid is to human leukemia cell line K562 cells;20 μ g/ml are to mouse
Myeloma cell SP20 have an obvious inhibiting effect, cell disruption is dead.The results show that bear gall powder, which has, inhibits tumour cell life
Effect that is long, extending life cycle, improve immune function.There is research by being ground to transplanted tumor cell H22 models in Mice Body
Study carefully, finds each dosage group of bear gall powder to the tumour inhibiting rate of Murine Hepatoma22 36.00% or more;Each dosage group is to H22 hepatoma mice
Weight, thymus index, spleen index and liver index all have no significant effect.The results show that bear gall powder to rat liver cancer H22 have it is bright
Aobvious inhibiting effect.
The effect of the inhibition thrombosis of bear gall powder pharmacologically.There is research by being formed to rat suppository and to blood platelet
It is demonstrated experimentally that after injection refined bear gall powder, can inhibition thrombosis, hence it is evident that inhibit the exception of hemorheological property, while to blood
Platelet aggregation also has certain inhibiting effect.There is apparent inhibition to make for the activation of blood platelet when refined bear gall powder is to cerebral infarction
With.Studies have found that bear bile powder for injection can obviously inhibit the formation of rat inside and outside thrombus, blood viscosity is made to reduce, pressed down
Platelet aggregation processed improves thrombotic ischemic tissue of brain lesion degree, reduces malonaldehyde level in damage brain tissue, protects super oxygen
Compound dismutase activity has protection and therapeutic effect to cerebral ischemia.
The protective effect to cell hypoxia damage of bear gall powder pharmacologically.There is research to be caused using sodium dithionite
ECV304 cells obviously damage.The results show that anoxic can obviously induce damage, Tauro ursodesoxy cholic acid caused by ECV304 cells
The cellular damage is can obviously reduce with Taurochenodeoxycholic Acid, there is significant anti-ECV304 cell hypoxias damage.Bear gall
Powder makes damaging cells number substantially reduce, and LDH leakages rate, cell mortality are remarkably decreased, and cell survival rate significantly improves.Its brain is protected
It is related to the protective effect of vascular endothelial cell anoxia-induced apoptosis with it to protect mechanism of action.
Bear gall powder anti-inflammatory, antibacterial pharmacologically and antivirus action.There is research by making mouse, rat wound, finds
Bear gall pearl acne cream obviously accelerates mouse soft-tissue trauma healing, substantially reduced to mouse ear edge swelling, makes diagonal
Rat toes swelling caused by pitching dish is substantially reduced, it was demonstrated that bear gall pearl acne cream has preferable anti-inflammatory effect.There is research to send out
Existing, bear gall powder has apparent bacteriostasis to staphylococcus aureus and Escherichia coli.Studies have found that 6 kinds of diseases of bear gall powder pair
Poison has inhibiting effect, wherein effect is significantly inhibited to sick 4 types (CoxB4) of coxsackie B race and parainfluenza virus I types (HVJ), it is right
Mouse influenza pneumonia significantly inhibits effect, causes dead mouse to have significant protective effect influenza.
The anti-allergic effects of bear gall powder pharmacologically.There is research to pass through IgE induced rats passive cutaneous anaphylaxis (PCA)
Experiment finds that refined bear gall powder can obviously inhibit P of Rats CA, can obviously inhibit release, the cell of mast cell degranulation histamine
Interior calcium intake and tumor necrosis factor-alpha, interleukin-6 and NF- κ B ρ 65, protein expression.Refined bear gall powder anti-I type is abnormal
One of important mechanisms of reaction are by inhibiting the expression of NF- kB proteins, to prevent the inflammatory cytokines such as TNF-α, IL-6
It generates.
The antifatigue effect of bear gall powder pharmacologically.Studies have found that when compound bear gall preparation may be swum by extending
Between and the pole-climbing time improve exercise tolerance, so that liver glycogen content is improved, blood urea nitrogen and Serum lactic acid content made to reduce, keep blood newborn
Acidohydrogenase vigor improves, and to provide more energy for body, has delayed the generation of fatigue, and then improve to exercise load
Adaptability;Compound bear gall preparation can be substantially reduced a shrinkage amplitude for rabbits' isolated duodenum smooth muscle, be to pass through
The contraction for inhibiting muscle excessive releases the spasm of smooth muscle to achieve the purpose that delay fatigue generates.
Bear gall powder antibechic pharmacologically, eliminating the phlegm, antipyretic, analgesic activity.There is research studies have shown that bear gall fritillary capsule energy
Enough substantially reduce concentrated ammonia liquor stimulation induced mice, cavy cough number;Promote mouse tracheae excretion phenol red and increases rat row
The effect of amount of expectoration;The experiment of rat fever caused by yeast powder also indicates that it with refrigeration function.The results show that bear gall fritillaria cough-relieving glue
Capsule has significant antibechic, eliminating the phlegm and faint refrigeration function.Studies have found that bear gall powder can significantly reduce 2,4- dinitros
Rat fever caused by phenol has certain refrigeration function, has apparent analgesic activity to pain caused by hot plate and acetic acid.
Bear gall powder has clinically been applied to ophthalmology disease.There is research to 92 acute bacterial conjunctivitis patients using single
Blind random grouping, parallel control method, observe curative effect and adverse reaction.Compound bear guts gutta combines compound neomycin sulphate drop
Ocular fluid 46 is treatment group, and Compound Neomycin Sulfate Eye Drops 46 are control group.As a result, effective 38 for the treatment of group, improvement 6
Example, obvious effective rate 82.6%, total effective rate 95.7%;Effective 29 of control group improves 8, and obvious effective rate 63.0% always has
Efficiency is 80.4%.As a result, compound bear guts gutta joint Compound Neomycin Sulfate Eye Drops are used for acute bacterial conjunctivitis,
It is curative for effect.There is research to 36 patients, takes bear gall powder 1g, addition normal saline solution is appropriate, is applied to eyelid bleb office outside
Portion.After 1~2 day, local pain disappears general coating, red and swollen substantially reduced, and scar is not stayed in bleb drying incrustation after decrustation.Knot
Fruit proves that bear gall treated powder eyelid Herpes Zoster is definite.
Bear gall powder has clinically been applied to angiocardiopathy.Bear gall pills for curing heart disease is mainly used in coronary heart diseases and angina pectoris etc.
The treatment of angiocardiopathy.There is research that the patient of 558 angiocardiopathies is randomly divided into treatment group and control group, treatment group
419, take orally bear gall pills for curing heart disease treatment;Control group 139, with suxiao jiuxin pills oral medication.Pass through electrocardiogram clinical observation
Treatment group 419, effective 138, effective 194, total effective rate be 79.24% control group 139, effective 47, effective 56
Example, total effective rate 74.10%.The results show that bear gall pills for curing heart disease is significant in efficacy to angiocardiopathy.
Bear gall powder has clinically been applied to respiratory disease.There is research by 84 Bronchopneumonia patients (age 2
Month~10 years old) it is divided into treatment group and control group, treatment group 42, using tanreqin injection (radix scutellariae, bear gall powder, cornu caprae hircus, gold
Honeysuckle flower and Fructus Forsythiae) added with antibiotic combination therapy;Control group 42, antibiotics alone treatment.Two groups of courses for the treatment of are 7~10 days.Knot
Fruit, treatment group's total effective rate 95.2%, control group total effective rate 71.4%.The results show that tanreqin injection treats bronchus
Pneumonia is significant in efficacy.Have research by 76 Patients With Respiratory Tract Infections (6 months ages~3 years old 52,3~7 years old 13,7~14 years old
11) it is randomly divided into treatment group and control group, treatment group 38 is added using by 0.25~4mL/kg/d of tanreqin injection
Intravenous infusion in 5% Glucose Liquid 100mL;5% Glucose Liquid is added using by Shuanhuanglian injection 1mL/kg/d in control group 38
Intravenous infusion in 100mL, two groups are treated 7 days.As a result, treatment group's total effective rate 94.74%, control group total effective rate 65.79%.Knot
Fruit proves that tanreqin injection treatment infantile respiratory tract infection is safe and effective, is worth extensive use.
Bear gall powder has clinically been applied to hemorrhoid.There is curative effect of the research for observation compound bear gall hemorrhoid frost, by selected disease
Example is randomly divided into treatment group and control group, treatment group 97, internal piles 45, external piles 24, mixed hemorrhoid 28;Control group 68,
Internal piles 38, external piles 12, mixed hemorrhoid 18.Treatment group's externally used compound bear gall hemorrhoid frost, control group is with 1:5000KMnO4 takes advantage of
Hip bath after heat is first smoked.As a result, the total effective rate for the treatment of group's internal piles, external piles, mixed hemorrhoid is respectively 97.78%, 95.83%,
96.43%;Control group internal piles, external piles, mixed hemorrhoid total effective rate be respectively 84.21%, 66.67%, 61.11%, display is multiple
The curative effect of square bear gall hemorrhoid frost is notable.
The present inventor is randomly divided into two groups in some clinical tests, by 242 cholecystitis and cholelithiasis patients:The present invention is real
Apply 1 gained bear gall powder of example (it may be simply referred to as gloomy precious bear gall powder in the present invention) treatment group and invigorating gallbladder and removing gall-stone particle control group.Its
Middle treatment group 187, control group 55, treatment group take orally bear gall powder (200mg, 2 times a day) 60, the oral profit of control group
Courage PAISHI KELI (meeting Chinese Pharmacopoeia one invigorating gallbladder and removing gall-stone particle specification of page 971 of version in 2015, once 2 bags, 2 times a day) 60
Day.As a result, treatment group cures 31, effective 80, effective 62, invalid 14, total effective rate is up to 92.5%.Treatment group treats
For cholecystolithiasis effective percentage up to 89.7%, treatment cholecystitis effective percentage is 95.6%.Above-mentioned clinical observation finds confirmation, bear of the present invention
Courage treated powder cholecystitis, gall stone are significant in efficacy.
Bear gall powder has clinically been applied to disease in the liver and gallbladder.There is research to use bear gall Capsule in Treating Chronic Hepatitis B
(CHB) 33, same period inpatient 20 is used in combination to compare, observes two groups of patient's jaundice recession situations.As a result, plus with bear gall glue
Capsule treatment can promote jaundice prompt resolution, liver function to be improved, and have no adverse effects to blood picture and renal function.There is research to report
Road Ursofalk treats chronic hepatitis B, and obstinate type jaundice has obvious curative effects.The present inventor refers to Liu Zhen justice document (Liu
Justice of shaking etc., Beijing traditional Chinese medicine, the 3rd phase of volume 35 in March, 2016), in some clinical tests, for 68 average ages 51.3
The chronic hepatitis B patient (male 48, female 20) in year, their liver fibrosis severity is:S1 phases 18, s2 phases 23
Example, S3 phases 14, S4 phases 13;Take 1 gained bear gall powder of the embodiment of the present invention (200mg, 2 times a day) 6 months daily;
Experiment is front and back to measure Fibrosis Indexes:Hyaluronic acid (HA), laminin (LN), type III precollagen (III-PC),
IV Collagen Type VIs (IV-C) as a result show 68 patients, four index averages from being respectively after becoming treatment before treating:243.6mg/L
Become 75.2mg/L, 265.3 μ g/L become 109.8 μ g/L, 183.2 μ g/L become 91.3 μ g/L, 103.7 μ g/L become 57.6 μ
G/L, each group treatment HA and LN have highly significant difference (p before and after treatment<0.01), III-PC and IV-C have significance difference before and after treatment
Different (p<0.05);In terms of clinical efficacy:Effective 27 (39.7%), it is effective 38 (55.9%), it is invalid 3 (4.4%), always have
Efficiency 95.6%.68 pretherapy and post-treatment Chinese medicine syndrome integrals of patient compare:Chinese medicine disease card integral (31.43 ± 5.68) before treatment
Point, it is (9.74 ± 4.35) point, the statistically significant (P of difference after treatment<0.05).
Common neoplastic hematologic disorder includes mainly each quasi-leukemia, Huppert's disease and malignant lymphoma.Acute white blood
Disease accounts for the 8th of common cancer, and lymthoma is also in top ten, and incidence increases year by year, Huppert's disease
Entire incidence accounts for 10% inside hematologic malignancies.Hesperian statistics show this current incidence of three classes tumour
All come the top ten of malignant tumour.
Bear gall powder provided by the invention is presented excellent performance and excellent physicochemical property is for example presented;In addition, the present invention carries
Excellent medical and health care function is presented in the bear gall powder of confession, such as described herein, can more effectively adjust blood fat, drop
Low total cholesterol, moreover it is possible to be more efficiently used for preventing the special efficacies such as cardiovascular and cerebrovascular disease, prevention of arterial atherosis, particularly
Ground can also be more efficiently used for preventing or treating gall stone disease and improve gallbladder function, more particularly can also more have
Effect ground is for hepatopathy and liver fibrosis and improves liver function;It can also more particularly be more efficiently used for preventing or treatment entity is swollen
Tumor and hematological system tumor;It can also be more particularly more efficiently used for inhibition thrombosis, inhibit platelet aggregation, prevent brain
Ischemic and ischemic cerebral disease;It is tired that raising immunity of organisms, raising muscle power, alleviation body can also be more particularly more efficiently used for
Labor.These technique effects are confirmed in the description of the present invention.
Specific implementation mode
The present invention can be further described by the following examples, however, the scope of the present invention and unlimited
In following embodiments.One of skill in the art, can be with it is understood that under the premise of without departing substantially from the spirit and scope of the present invention
Various change and modification are carried out to the present invention.The present invention carries out the material and test method that are arrived used in experiment general
And/or specific description.Although to realize the present invention many materials and operating method used in purpose be it is known in the art that
But the present invention is still described in detail as far as possible herein.
The assay method of hydroscopicity and moisture:Test sample 2g is taken, is laid in drying to the flat weighing bottle of constant weight, it is thick
Degree is no more than 5mm, accurately weighed;It is 5 hours dry at 100~105 DEG C to open bottle cap, bottle cap is covered, it is cold in dislocation drier
But 30 minutes, accurately weighed weight;It is dried 1 hour in above-mentioned temperature again, it is cooling, it weighs, until the difference weighed twice in succession is not
Until 5mg.According to the weight of less loss, the percentage containing moisture in test sample is calculated, the hydroscopicity and water of sample are calculated
Divide content.
Embodiment 1:Prepare bear gall powder
(1) the bear gall juice by the acquisition of no Tube Drain method filters, and obtains filtrate, adds the water dilution of 0.75 times of amount, obtains bear bile fluid,
12 hours are stood at 2~6 DEG C of temperature, then carries out subsequent operation;
(2) in a manner of bubbling, it is filled with nitrogen 27min into bear bile fluid obtained by step (1), is then filled with carbon dioxide
18min;
(3) bear bile fluid handled through aforesaid operations is inclined into material disc (1.5~2.5cm of liquid layer depth), be immediately placed on
It is refrigerated in -50 DEG C of freeze drier, freezes 3.5 hours in advance;Then -22 DEG C are warming up to 0.3 DEG C of speed per minute,
It is kept for 1 hour;Then -3 DEG C are warming up to 0.3 DEG C of speed per minute, are kept for 2.5 hours;Then with 0.3 DEG C of speed per minute
Degree is warming up to 10 DEG C, is kept for 4.5 hours;Then 32 DEG C are warming up to 0.3 DEG C of speed per minute, are kept for 2.5 hours, terminated cold
Freeze drying program;
(4) by the dried lump in material disc be ground into can by the fine powder that 50 mesh sieve to get.
Bear gall powder obtained by the present embodiment is the burnished gold or bright-yellow powder of hyaloid gloss, shows bright orange at 365nm
Color fluorescence,
Hydroscopicity≤2% (0.30%),
Solubility in water (13.97g/100ml) within the scope of 13.6~14.2g/100ml,
Solubility in 90% ethyl alcohol within the scope of 1.25~2g/100ml (1.84g/100ml),
The pH value of 5% aqueous solution (pH value 6.65) in 6.0~7.0 ranges,
Moisture is less than 3% (0.62%).
Embodiment 2:Prepare bear gall powder
(1) the bear gall juice by the acquisition of no Tube Drain method filters, and obtains filtrate, adds the water dilution of 0.5 times of amount, obtains bear bile fluid,
12 hours are stood at 2~6 DEG C of temperature, then carries out subsequent operation;
(2) in a manner of bubbling, it is filled with nitrogen 30min into bear bile fluid obtained by step (1), is then filled with carbon dioxide
15min;
(3) bear bile fluid handled through aforesaid operations is inclined into material disc (1.5~2.5cm of liquid layer depth), be immediately placed on
It is refrigerated in -55 DEG C of freeze drier, freezes 3 hours in advance;Then -23 DEG C are warming up to 0.3 DEG C of speed per minute, protected
It holds 1.5 hours;Then -2 DEG C are warming up to 0.25 DEG C of speed per minute, are kept for 2 hours;Then with 0.25 DEG C of speed per minute
Degree is warming up to 12 DEG C, is kept for 5 hours;Then 30 DEG C are warming up to 0.25 DEG C of speed per minute, are kept for 3 hours, terminate freezing
Drying program;
(4) by the dried lump in material disc be ground into can by the fine powder that 50 mesh sieve to get.
Bear gall powder obtained by the present embodiment is the burnished gold or bright-yellow powder of hyaloid gloss, shows bright orange at 365nm
Color fluorescence,
Hydroscopicity≤2% (0.51%),
Solubility in water (14.06g/100ml) within the scope of 13.6~14.2g/100ml,
Solubility in 90% ethyl alcohol within the scope of 1.25~2g/100ml (1.34g/100ml),
The pH value of 5% aqueous solution (pH value 6.26) in 6.0~7.0 ranges,
Moisture is less than 3% (0.49%).
Embodiment 3:Prepare bear gall powder
(1) the bear gall juice by the acquisition of no Tube Drain method filters, and obtains filtrate, adds the water dilution of 1 times of amount, bear bile fluid is obtained, 2
12 hours are stood at~6 DEG C of temperature, then carries out subsequent operation;
(2) in a manner of bubbling, it is filled with nitrogen 25min into bear bile fluid obtained by step (1), is then filled with carbon dioxide
20min;
(3) bear bile fluid handled through aforesaid operations is inclined into material disc (1.5~2.5cm of liquid layer depth), be immediately placed on
It is refrigerated in -45 DEG C of freeze drier, freezes 4 hours in advance;Then -20 DEG C are warming up to 0.35 DEG C of speed per minute,
It is kept for 1.2 hours;Then -2 DEG C are warming up to 0.35 DEG C of speed per minute, are kept for 3 hours;Then with 0.34 DEG C per minute
Speed is warming up to 15 DEG C, is kept for 4 hours;Then 35 DEG C are warming up to 0.35 DEG C of speed per minute, are kept for 2 hours, terminated cold
Freeze drying program;
(4) by the dried lump in material disc be ground into can by the fine powder that 50 mesh sieve to get.
Bear gall powder obtained by the present embodiment is the burnished gold or bright-yellow powder of hyaloid gloss, shows bright orange at 365nm
Color fluorescence,
Hydroscopicity≤2% (0.44%),
Solubility in water (13.73g/100ml) within the scope of 13.6~14.2g/100ml,
Solubility in 90% ethyl alcohol within the scope of 1.25~2g/100ml (1.35g/100ml),
The pH value of 5% aqueous solution (pH value 6.42) in 6.0~7.0 ranges,
Moisture is less than 3% (0.54%).
Embodiment 4:Prepare bear gall powder
(1) the bear gall juice by the acquisition of no Tube Drain method filters, and obtains filtrate, adds the water dilution of 0.6 times of amount, obtains bear bile fluid,
12 hours are stood at 2~6 DEG C of temperature, then carries out subsequent operation;
(2) in a manner of bubbling, it is filled with nitrogen 30min into bear bile fluid obtained by step (1), is then filled with carbon dioxide
10min;
(3) bear bile fluid handled through aforesaid operations is inclined into material disc (1.5~2.5cm of liquid layer depth), be immediately placed on
It is refrigerated in -52 DEG C of freeze drier, freezes 2 hours in advance;Then -23 DEG C are warming up to 0.25 DEG C of speed per minute,
It is kept for 1.5 hours;Then it is warming up to 0 DEG C with 0.3 DEG C of speed per minute, is kept for 2.3 hours;Then with 0.24 DEG C per minute
Speed is warming up to 13 DEG C, is kept for 4.2 hours;Then 32 DEG C are warming up to 0.27 DEG C of speed per minute, are kept for 2.5 hours, knot
Beam is freeze-dried program;
(4) by the dried lump in material disc be ground into can by the fine powder that 50 mesh sieve to get.
Bear gall powder obtained by the present embodiment is the burnished gold or bright-yellow powder of hyaloid gloss, shows bright orange at 365nm
Color fluorescence,
Hydroscopicity≤2% (0.52%),
Solubility in water (13.68g/100ml) within the scope of 13.6~14.2g/100ml,
Solubility in 90% ethyl alcohol within the scope of 1.25~2g/100ml (1.92g/100ml),
The pH value of 5% aqueous solution (pH value 6.34) in 6.0~7.0 ranges,
Moisture is less than 3% (0.45%).
Embodiment 5:Prepare bear gall powder
(1) the bear gall juice by the acquisition of no Tube Drain method filters, and obtains filtrate, adds the water dilution of 0.8 times of amount, obtains bear bile fluid,
12 hours are stood at 2~6 DEG C of temperature, then carries out subsequent operation;
(2) in a manner of bubbling, it is filled with nitrogen 20min into bear bile fluid obtained by step (1), is then filled with carbon dioxide
20min;
(3) bear bile fluid handled through aforesaid operations is inclined into material disc (1.5~2.5cm of liquid layer depth), be immediately placed on
It is refrigerated in -48 DEG C of freeze drier, freezes 5 hours in advance;Then -23 DEG C are warming up to 0.32 DEG C of speed per minute,
It is kept for 1 hour;Then -5 DEG C are warming up to 0.35 DEG C of speed per minute, are kept for 2.6 hours;Then with 0.33 DEG C per minute
Speed is warming up to 12 DEG C, is kept for 4.8 hours;Then 34 DEG C are warming up to 0.34 DEG C of speed per minute, are kept for 2.8 hours, knot
Beam is freeze-dried program;
(4) by the dried lump in material disc be ground into can by the fine powder that 50 mesh sieve to get.
Bear gall powder obtained by the present embodiment is the burnished gold or bright-yellow powder of hyaloid gloss, shows bright orange at 365nm
Color fluorescence,
Hydroscopicity≤2% (0.49%),
Solubility in water (13.91g/100ml) within the scope of 13.6~14.2g/100ml,
Solubility in 90% ethyl alcohol within the scope of 1.25~2g/100ml (1.75g/100ml),
The pH value of 5% aqueous solution (pH value 6.54) in 6.0~7.0 ranges,
Moisture is less than 3% (0.80%).
The method for respectively referring to embodiment 1- embodiments 5, different be only be not diluted with water in step (1), but directly
Bear gall juice stoste is subjected to subsequent operation, thus obtained five batches of samples are presented dark yellow or brown color, have no hyaloid gloss,
Remaining parameter includes solubility in solubility, ethyl alcohol in hydroscopicity, water, 5 gained of pH value, moisture and embodiment 1- embodiments
Bear gall powder is essentially identical.This shows to dodge in freeze-drying plus a small amount of water contributes to improvement product appearance, obtains being in hyaloid gloss
Powder, may infer that bear gall powder of the present invention due to being added to water form ice crystal, bear gall powder of the present invention after advance deep freeze
Also referred to as ice crystal bear gall powder or ice crystal state bear gall powder.
Embodiment 11:Prepare bear gall powder
The method for respectively referring to embodiment 1- embodiments 5, different is only not inflated with nitrogen in step (2), obtains five batches of bears
Courage powder.
Bear gall powder obtained by the present embodiment is light yellowish brown powder, and glassy yellow fluorescence is shown at 365nm,
Hydroscopicity≤2% (0.84~1.35%),
Solubility in water within the scope of 8.6~9.2g/100ml,
Solubility in 90% ethyl alcohol within the scope of 0.83~0.96g/100ml,
The pH value of 5% aqueous solution in 6.2~6.7 ranges,
Moisture is less than 3% (0.82~1.03%).
Embodiment 12:Prepare bear gall powder
The method for respectively referring to embodiment 1- embodiments 5, different is only not filling CO 2 in step (2), obtains five
Criticize bear gall powder.
Bear gall powder obtained by the present embodiment is light yellowish brown powder, and glassy yellow fluorescence is shown at 365nm,
Hydroscopicity≤2% (0.81~1.31%),
Solubility in water within the scope of 8.8~9.8g/100ml,
Solubility in 90% ethyl alcohol within the scope of 0.87~0.93g/100ml,
The pH value of 5% aqueous solution in 6.2~6.7 ranges,
Moisture is less than 3% (0.88~1.03%).
Embodiment 13:Prepare bear gall powder
The method for respectively referring to embodiment 1- embodiments 5, different is only that inflated with nitrogen does not also fill titanium dioxide in step (2)
Carbon but directly carry out next step, obtain five batches of bear gall powders.
Bear gall powder obtained by the present embodiment is light yellowish brown powder, and glassy yellow fluorescence is shown at 365nm,
Hydroscopicity≤2% (0.94~1.21%),
Solubility in water within the scope of 9.3~9.7g/100ml,
Solubility in 90% ethyl alcohol within the scope of 0.94~0.98g/100ml,
The pH value of 5% aqueous solution in 6.26~6.75 ranges,
Moisture is less than 3% (0.84~1.03%).
In addition, obtained from commercially available approach five batches of bear gall powders (Chinese medicines quasi-word Z19991061, Z20000030,
Z20000026, Z20000055, Z20000031, wherein Z20000026 bear gall powders can may be simply referred to as Z26 or Z26 bears in the present invention
Courage powder, other four batches can also state in a similar way), their related physicochemical property parameter is measured, it is as a result as follows:Hydroscopicity≤
2% (in 0.57~1.73% range), solubility in water is within the scope of 8.3~10.1g/100ml, in 90% second
Solubility in alcohol is within the scope of 0.84~1.17g/100ml, and the pH value of 5% aqueous solution is in 6.0~7.0 ranges, moisture
Content is less than 3% (in 0.51~1.66% range).
Bear gall powder is prepared using the method recorded in CN102114044B its [0180] to [0183], measures their correlation
Physicochemical property parameter is as a result as follows:Hydroscopicity 1.42%, solubility 9.7g/100ml in water are molten in 90% ethyl alcohol
Solution degree 0.82g/100ml, the pH value 6.4 of 5% aqueous solution, moisture 1.47%;This batch of bear gall powder is known as in the present invention
044#。
Bear gall powder is prepared using the method recorded in CN105147729A its [0022] to [0024], measures their correlation
Physicochemical property parameter is as a result as follows:Hydroscopicity 1.17%, solubility 8.9g/100ml in water are molten in 90% ethyl alcohol
Solution degree 0.68g/100ml, the pH value 6.7 of 5% aqueous solution, moisture 1.26%;This batch of bear gall powder is known as in the present invention
729#。
The method recorded in specification " alcohol extraction procedure one " using CN1311002A prepares bear gall powder, measures them
Related physicochemical property parameter, it is as a result as follows:Hydroscopicity 1.86%, solubility 10.2g/100ml in water, in 90% ethyl alcohol
In solubility 0.91g/100ml, the pH value 6.6 of 5% aqueous solution, moisture 1.37%;This batch of bear gall powder is in the present invention
Referred to as 002#.
From result above as it can be seen that bear gall powder made from method of the embodiment of the present invention 1 to embodiment 5 shows notable higher
Solubility.
Test example 1:The quality testing of bear gall powder
1, character:Embodiment 1 to 5 each sample of embodiment is the burnished gold or bright-yellow powder of hyaloid gloss;Bitter,
Micro- raw meat.
Embodiment 11 to 13 each sample of embodiment and five batches of commercially available bear gall powders, 044#, 729#, 002# is light yellowish brown
Powder;Bitter, micro- raw meat.
2, differentiate (1):It takes this product content appropriate (being equivalent to bear gall powder 0.06g), ethyl alcohol 5ml is added to make dissolving, filter, filter
Liquid is evaporated, and residue adds 10% sodium hydroxide solution 5ml, is set in water-bath heating hydrolysis 8 hours (or 120 DEG C hydrolyze 2 hours), is put
It is cold, salt acid for adjusting pH value is added dropwise to 2~3, with ethyl acetate shaking extraction 2 times, each 10ml, combined ethyl acetate liquid is evaporated,
Residue adds ethyl alcohol 5ml to make dissolving, stands, takes supernatant as test solution.Separately take ursodesoxycholic acid reference substance, chenodeoxycholic
Sour reference substance and cholic acid reference substance add ethyl alcohol that the 1ml respectively mixed solutions containing 0.5mg are made, as a contrast product solution.According to thin layer color
Spectrometry (four general rules 0502 of Chinese Pharmacopoeia version in 2015) is tested, and is drawn each 4ul of above two solution, is put respectively in same silica gel
On G lamellaes, with isooctane-isoamyl ether-n-butanol-glacial acetic acid-water (10:5:3:5:1) upper solution (face with prepare) is
Solvent is unfolded, and takes out, dries, and with 10% ethanol solution of sulfuric acid, it is clear to be heated to spot development at 105 DEG C, sets ultraviolet light for spray
It is inspected under lamp (365nm).It is general to require, in test sample chromatography, on position corresponding with reference substance chromatography, show same color
Fluorescence spot.As a result:Embodiment 1 is to 5 each sample of embodiment, 11 embodiment of embodiment, 13 each sample and five batches of commercially available bear galls
Powder, 044#, 729#, 002# show the fluorescence spot of same color on position corresponding with reference substance chromatography.
3, differentiate (2):Test sample is taken, according to the method test under this test example (assay) item.It is general to require, test sample
In chromatography, chromatographic peak identical with reference substance main peak retention time should be presented.As a result:Embodiment 1 is to 5 each sample of embodiment, reality
It applies 11 embodiment of example, 13 each sample and five crowdes of commercially available bear gall powders, 044#, 729#, 002# is presented when retaining with reference substance main peak
Between identical chromatographic peak.
4, Check pig galls are examined:Hyodesoxycholic acid reference substance is taken, adds ethyl alcohol 1ml the is made solution containing 0.5mg, product are molten as a contrast
Liquid.It is tested according to thin-layered chromatography (four general rules 0502 of Chinese Pharmacopoeia version in 2015), draws the test solution differentiated under (1) item
With each 4ul of above-mentioned reference substance solution, put respectively on same silica gel g thin-layer plate, with isooctane-isoamyl ether-n-butanol-ice vinegar
Acid-water (10:5:3:5:1) upper solution (facing with preparation) is solvent, is unfolded, takes out, dry, sprays with 10% sulfuric acid ethyl alcohol
Solution, it is clear to be heated to spot development at 105 DEG C, sets and is inspected under ultraviolet lamp (365nm).It is general to require, in test sample chromatography,
On position corresponding with reference substance chromatography, the fluorescence spot of same color should must not be shown.As a result:Embodiment 1 is each to embodiment 5
The not aobvious same color of sample, 11 embodiment of embodiment, 13 each sample and five crowdes of commercially available bear gall powders, 044#, 729#, 002#
Fluorescence spot.
5, assay
It is measured according to high performance liquid chromatography (four general rules 0512 of Chinese Pharmacopoeia version in 2015).
Chromatographic condition and system suitability:Using octadecylsilane chemically bonded silica as filler;With methanol-di(2-ethylhexyl)phosphate
Hydrogen sodium solution (0.03mol/L) (68:32) (being 4.4 with phosphorus acid for adjusting pH value) is mobile phase;Detection wavelength is 210nm;Column temperature
It is 40 DEG C.Number of theoretical plate is calculated by Tauro ursodesoxy cholic acid peak should be not less than 2500.
The preparation of reference substance solution:Take taurine sodium deoxycholate reference substance appropriate, it is accurately weighed, add methanol to be made often
Solution of the 1ml containing 1mg is to get (being equivalent to Tauro ursodesoxy cholic acid 0.9578mg).
The preparation of test solution:This product content under content uniformity item, finely ground, mixing is taken to take and (be equivalent to bear in right amount
Courage powder 0.12g), it is accurately weighed, it sets in 50ml measuring bottles, adds methanol appropriate, 10 points of supersound process (power 300W, frequency 50kHz)
Clock is let cool, and with methanol dilution to scale, is shaken up, filtration, take subsequent filtrate to get.
Measuring method:It is accurate respectively to draw reference substance solution and each 5~10ul of test solution, liquid chromatograph is injected, is surveyed
It is fixed to get.
General to require, the Tauro ursodesoxy cholic acid (C26H45NO6S) in bear gall powder must not be less than 30%.
As a result:Tauro ursodesoxy cholic acid (C26H45NO6S) content in embodiment 1 to 5 each sample of embodiment 35~
Tauro ursodesoxy cholic acid (C26H45NO6S) content in 38% range, such as in embodiment 1 is 37.3%;Embodiment 11 is to reality
Apply the Tauro ursodesoxy cholic acid in 13 each sample of example and five batches of commercially available bear gall powders, 044#, 729#, 002# each sample
(C26H45NO6S) for content in 23~32% ranges, some contents are more than 30%, somewhat smaller than 30%, such as embodiment 11
Tauro ursodesoxy cholic acid (C26H45NO6S) content in the bear gall powder with reference to obtained by 1 method of embodiment is 27.0%.
The above result shows that present invention gained bear gall powder has the typical activity substance Tauro ursodesoxy cholic acid of high level.
Test example 2:The stability of bear gall powder
It is protected from light and is sealed 6 months under the conditions of so that bear gall powder sample is placed in 40 DEG C of constant humidity, 0 is measured according to the method for test example 1
Tauro ursodesoxy cholic acid content when the moon and June in sample, criticizes sample to Mr. Yu, its sample after this high-temperature treatment is calculated according to following formula
Tauro ursodesoxy cholic acid remnants rates in product:
Remaining rate %=[0 month Tauro ursodesoxy cholic acid content of Tauro ursodesoxy cholic acid content ÷ in June] × 100%
As a result:Tauro ursodesoxy cholic acid remnants rates in embodiment 1 to 5 each sample of embodiment are in 97~100% ranges
It is interior, such as 1 sample Tauro ursodesoxy cholic acid remnants rates of embodiment are 99.4%;Ox in embodiment 11 to 13 each sample of embodiment
Sulphur ursodesoxycholic acid remnants rates are in 88~91% ranges, such as the bear gall powder sample with reference to obtained by 1 method of embodiment of embodiment 11
Product Tauro ursodesoxy cholic acid remnants rates are 90.4%;Ox sulphur bear in five batches of commercially available bear gall powders, 044#, 729#, 002# each sample
Deoxycholic aicd remnants rates are in 87~93% ranges, such as 729# bear gall powder sample Tauro ursodesoxy cholic acid remnants rates are
89.6%.These are the result shows that present invention gained bear gall powder has more excellent chemical stability.
Test example 3:Bear gall powder adjusts blood fat experimental study
In recent years, China resident suffers from hyperlipidemia number and constantly rises, and is in rejuvenation trend, this is moved with diet structure
Physical property food intake dose increases, and the opposite reduction of plant food intake is related.Medical research confirms that hyperlipidemia is to induce to move
The important risk factor of pulse atherosclerosis and cardiovascular and cerebrovascular disease.Prevention to hyperlipidemia, to control cardiovascular and cerebrovascular disease
It is of great importance.Method disclosed in the flat document of reference cell jasmine (straits preventive medicine magazine, the 2nd phase of volume 13 in 2007),
Measure the effect of the adjusting blood fat of bear gall powder.
Experiment is respectively with bear gall powder:1 bear gall powder of embodiment, 2 bear gall powder of embodiment, 3 bear gall powder of embodiment, embodiment 13
Bear gall powder, Z26 bear gall powders, 002# bear gall powders with reference to obtained by 1 method of embodiment.
It is 1g that bear gall powder, which recommends Coming-of-Age Day dose, and it is per kilogram of body weight 20mg to be converted to daily intaking amount, is recommended by human body
12 times of setting administration groups of amount, i.e. 120mg/kg separately set blank control group, control group high in fat, and each group animal is grouped at random, every group
It 12, is administered once a day, totally 30 days, measures three blood lipid levels after experiment in accordance with the law.The results show that blank control group, high in fat
Changes of weight between control group, each medicine-feeding test group animal is essentially identical, no significant difference.
Influence to animal TC:After bear gall powder 30d, blank control group TC (mean value 1.46mmol/L) is significantly lower than high in fat
Group (mean value 2.63mmol/L, P<0.01), indicate that model is set up;Administration group TC gives bear gall powder examination compared with control group high in fat
(p is remarkably decreased after sample<0.01), show that influence of the sample to TC indexs is the positive, bear gall powder is relatively given with control group high in fat
TC reduces percentage after sample, and each medicine-feeding test group result is respectively:1 bear gall powder of embodiment is 21.5%, 2 bear gall powder of embodiment
It is 20.5% for 22.8%, embodiment, 3 bear gall powder, the bear gall powder with reference to obtained by 1 method of embodiment of embodiment 13 is 10.8%, Z26
Bear gall powder is 11.2%, 002# bear gall powders are 9.7%.These are the result shows that embodiment 1-3 bear gall powders can than other bear gall powders
Significantly more reduce TC.
Influence to TG levels:After bear gall powder 30d, more high in fat group of blank control group TG (mean value 0.88mmol/L) is low
(mean value 1.41mmol/L, P<0.05), hints model is set up;Administration group TG gives bear gall powder sample compared with control group high in fat
After be remarkably decreased (p<0.01), show that influence of the sample to TG indexs is the positive, bear gall powder examination is relatively given with control group high in fat
TG reduces percentage after sample, and each medicine-feeding test group result is respectively:1 bear gall powder of embodiment is 39.9%, 2 bear gall powder of embodiment is
38.2%, 3 bear gall powder of embodiment is 41.7%, the bear gall powder with reference to obtained by 1 method of embodiment of embodiment 13 is 24.4%, Z26 bears
Courage powder is 26.7%, 002# bear gall powders are 24.1%.These are the result shows that embodiment 1-3 bear gall powders can be more than other bear gall powders
Significantly reduce TG.
Influence to HDL-C (high-density lipoprotein cholesterol):After bear gall powder 30d, the HDL-C of each medicine-feeding test group with
Control group high in fat is similar.Show that influence of the bear gall powder to Serum HDL-C index is feminine gender, medicine-feeding test group and control group high in fat
HDL-C is within the scope of 1.38~1.44mmol/L.
As it can be seen that bear gall powder can make animal TC (serum total cholesterol) and TG, (triglycerides reflects blood fat water to the studies above
It is flat) level reduction, compared with control group high in fat, difference is statistically significant.According to the Ministry of Public Health《Health food is examined and evaluation
Technical specification》Criterion judges that the animal functional experiment of bear gall powder has the function of adjusting blood fat, and also having, which reduces total courage, consolidates
The effect of alcohol.Lipid metabolism is closely related with cardiovascular and cerebrovascular disease, and cardiovascular and cerebrovascular disease is mostly derived from atherosclerosis, high blood
One of the main reason for fat is then initiation atherosclerosis (AS), especially TC levels increase, and are the Danger Indexes for causing AS.From
It is said in this meaning, bear gall powder of the present invention will be helpful to prevent cardiovascular and cerebrovascular disease and atherosclerosis.
Test example 4:Bear gall powder dissolves gall stone, improves gallbladder function and protects the effect test of liver function
1. test material
Animal:New zealand white rabbit, 2.5~3kg of weight, Guangdong Medical Lab Animal Center provide, animal licensing
Scxk (Guangdong) 2003-0002, check and affirm word 2007A001 in Guangdong.
Drug and reagent:
Methyl tertiary butyl ether:It analyzes pure.
People's cholesterol stone:Certain hospital provides, and the calculus containing 95% or more cholesterol is detected as through infrared spectrum.Serum creatinine
Testing cassete, uric acid testing cassete (biology is built up in Nanjing), urea ammonia testing cassete (middle raw north control), (Sichuan steps AST, ALT testing cassete
Gram), GGT testing cassetes (Jin Taier).
2. animal experiment
Healthy new zealand white rabbit, half male and half female is taken to be grouped at random by weight, every group 6, group is:
Blank control group (physiological saline),
Control group (methyl tertiary butyl ether),
Test group A (methyl tertiary butyl ether adds 1 bear gall powder of embodiment),
Experiment group B (methyl tertiary butyl ether adds the bear gall powder with reference to obtained by 1 method of embodiment of embodiment 13),
Test group C (methyl tertiary butyl ether adds Z26 bear gall powders),
Test group D (methyl tertiary butyl ether adds 002# bear gall powders).
The solution of 2g/100ml is made of water dissolution for bear gall powder, is used for animal experiment.
When experiment, to each experimental rabbit ear after intravenous injection 3% yellow Jackets anesthesia (1ml/kg), it is fixed on rabbit and fixes
On device, it is made to lie on the back, free tracheae of performing the operation goes forward side by side promoting the circulation of qi cannula to prevent its death by suffocation, from xiphoid-process median incision of lower abdomen five
Centimetre, it is successively segregated into abdominal cavity, gall-bladder is fully exposed, lifts gall-bladder with anodontia tissue clamps, with small scissors in gallbladder neck upper end
An osculum is cut, toward one gall stone for weighing up weight of merging in gall-bladder after waiting bile to empty, balloon catheter, suture are inserted into from notch
Opening.It is inserted into balloon catheter, suture opening with gallbladder neck notch.An osculum equally is opened at gall-bladder bottom, is sutured after being placed in conduit.It is past
Conduit saline injection after confirming suture ne-leakage, through 2 milliliters of gall-bladder bottom conduit saline injection, rinses gall-bladder.So
It is administered afterwards according to such as under type:
Control group 2 milliliters of saline injection in gall-bladder, it is primary every 30 minutes, continue 10 hours;
Methyl tertiary butyl ether group injects air after with normal saline flushing gall-bladder, toward gall-bladder, by the physiological saline in gall-bladder
Then 1 milliliter of 1 milliliter of methyl tertiary butyl ether and physiological saline are injected in discharge in gall-bladder, primary every perfusion in 30 minutes, continue 10
Hour;
Each drug study group injects air after with normal saline flushing gall-bladder, toward gall-bladder, by the physiological saline in gall-bladder
Then 1 milliliter of 1 milliliter of methyl tertiary butyl ether and bear gall powder solution are injected in discharge in gall-bladder, primary every perfusion in 30 minutes, continue
10 hours.
Above-mentioned animal clamps conducting pipes nozzle after the saline injection toward in gall-bladder or drug, with artery clamp, so that
It obtains gall-bladder and keeps expanded state, every breathing, the heartbeat of 30 minutes observation animals.After experiment, animal is put to death, courage is cut off
Capsule observes litholysis situation, calculates dissolve stone rate.Gall-bladder normal saline flushing is clean, observes gallbladder wall degree of impairment, and will
Nearby hepatic tissue is preserved with 10% formalin for gall-bladder and gall-bladder, makes slice, the degree of impairment of light microscopic observation gallbladder wall.On
It states before and after zoopery that arteria carotis takes blood, refrigerated centrifuge to detach serum respectively, measures Uric Acid Content, urea nitrogen, serum creatinine and contain
Amount, alanine aminotransferase activity, γ-glutamyl transferase active, aspartate aminotransferase activity and AST/
ALT values.
3. Testing index
(1) dissolve stone rate:After above-mentioned animal completes experiment, each experimental animal gall-bladder is cut, cuts off gall-bladder, takes out remaining calculus
And residue, with trickle irrigation plus physiological saline wash clean, drying is weighed.Dissolve stone rate calculates as follows:
Dissolve stone rate=(former calculus weight-residue calculus weight)/original calculus weight × 100%
(2) Biochemistry test:Above-mentioned animal experiment before and after take 2~4 milliliters of blood through arteria carotis respectively, freeze from
The heart detaches serum, is used for the detection of related biochemical indicator.
(3) blood urea nitrogen content detection:Detection method is 2 dynamic methods of enzyme, and sample is serum before and after animal experiment, is surveyed
It is fixed that steps are as follows:Normal concentration is 7.14mmol/L, measures wavelength:Dominant wavelength 340nm, commplementary wave length 405nm
After mixing, start within 30 seconds to measure absorbance A 1, absorbance A 2 was measured after 20~60 seconds.
It calculates:
Δ A standards=(A1-A2) standard-(A1-A2) blank
Δ A samples=(A1-A2) sample-(A1-A2) blank
Urea nitrogen concentration=Δ A samples/Δ A standards × normal concentration (mmol/L)
(4) Uric Acid Content measures:
Illustrate by kit, is detected with automatic biochemical analyzer.
(5) blood creatinine assay:
It is measured with blood creatinine assay kit, takes serum 0.2ml that two 2ml of reagent of kit configuration is added,
It mixes well, 3000rpm centrifuges 10min, and supernatant according to the form below is taken to be measured:
Measure pipe | Standard pipe | Blank tube | |
Haemocyanin filtrate/diluted urine (ml) | 1.6 | - | - |
Reagent one (10 μm of ol/L) flesh liver standard items (ml) | - | 1.6 | - |
Distilled water (ml) | - | - | 1.6 |
Reagent three (ml) | 0.5 | 0.5 | 0.5 |
Reagent four (ml) | 0.5 | 0.5 | 0.5 |
Mixing, 37 DEG C of water-bath 10min, flowing water cools down after taking-up, 1cm optical paths, the small length of 510nm, and distilled water zeroing measures each
Pipe absorbance value.
Calculation formula:
Serum flesh liver (μm ol/L)=measurement pipe absorbance × standard pipe concentration (10 μm of ol/L) × 11/ standard pipe absorbance
(6) blood AST Activity determinations:
It is measured with blood AST Activity determinations with kit, assay method is one-step method.
The preparation of working solution:R1 and R2 reagents, which are measured, according to reagent needed for measurement presses 4:The mixing of 1 ratio is working solution.It surveys
Standing wave is long:Dominant wavelength 340nm, commplementary wave length 405nm.
It measures (U) | It calibrates (C) | Working solution (B) | |
Sample (μ l) | 40 | - | - |
It calibrates (μ l) | - | 40 | - |
Distilled water (μ l) | - | - | 40 |
Working solution (μ l) | 1000 | 1000 | 1000 |
Mixing, 37 DEG C of water-bath 1min measure initial absorbance, then Accurate Determining average minute clock absorbance change rate Δ
A/min。
It calculates:Sample AST vigor (U/L)=Δ A/min × F (theoretical value=- 4180 F)
(7) blood ALT Activity determinations:
It is measured with blood ALT Activity determinations with kit, assay method is one-step method.
The preparation of working solution:R1 and R2 reagents, which are measured, according to reagent needed for measurement presses 4:The mixing of 1 ratio is working solution.It surveys
Standing wave is long:Dominant wavelength 340nm, commplementary wave length 405nm.
It measures (U) | It calibrates (C) | Working solution (B) | |
Sample (μ l) | 40 | - | - |
It calibrates (μ l) | - | 40 | - |
Distilled water (μ l) | - | - | 40 |
Working solution (μ l) | 1000 | 1000 | 1000 |
Mixing, 37 DEG C of water-bath 1min measure initial absorbance, then Accurate Determining average minute clock absorbance change rate Δ
A/min。
It calculates:Sample ALT vigor (U/L)=Δ A/min × F (theoretical value=- 4180 F)
(8) blood GGT Activity determinations:
It is measured with blood GGT Activity determinations with kit, detection method is performance rate method.
Measure maneuver《Standard methodology》It is standard with Hitachi 7250, measures wavelength:Dominant wavelength 405nm, commplementary wave length
505nm。
It calculates:
Sample concentration=sample absorbance × normal concentration/standard absorbance
4. morphological indexes
After experiment, with the clean gall-bladder of normal saline flushing, gallbladder wall situation is observed, is preserved in 10% formalin,
Do pathological section, light microscopic observation gallbladder wall degree of impairment.
Statistical method:One-way analysis of variance is used to the difference of the blood parameters detected before and after experiment, is compared two-by-two
It is examined compared with using q;The comparison of gallbladder wall operation uses rank sum test.Inspection level a=0.05.p<When 0.05, difference has statistics
Learning meaning, i.e. there were significant differences, p<When 0.1, statistically significant difference.
5. experimental result
(1) influence of gall stone dissolve stone rate is dissolved to methyl tertiary butyl ether --- prevent or treat the effect of gall stone disease
Remaining calculus is obtained after experiment, drying is weighed, and dissolve stone rate is calculated.Compared with physiological saline group, methyl- tert fourth
Ether group, methyl tertiary butyl ether add bear gall group to have raising, but lift degree is different, as a result see the table below.
Group | Dissolve stone rate (%) |
Blank control group (physiological saline) | 0.84±0.49 |
Control group (methyl tertiary butyl ether) | 38.63±5.66 |
Test group A (methyl tertiary butyl ether adds 1 bear gall powder of embodiment) | 85.32±10.62 |
Experiment group B (methyl tertiary butyl ether adds the bear gall powder with reference to obtained by 1 method of embodiment of embodiment 13) | 54.28±8.93 |
Test group C (methyl tertiary butyl ether adds Z26 bear gall powders) | 58.37±12.47 |
Test group D (methyl tertiary butyl ether adds 002# bear gall powders) | 49.57±9.41 |
Test group A significant difference, p compared with methyl tertiary butyl ether control group<0.05;Experiment group B, C, D and methyl- tert
Butyl ether control group compares variant, p<0.1;1 bear gall powder of embodiment is significantly better than other bear gall powders.
(2) gall-bladder pathologic finding --- improve and protect the effect of gallbladder function
Animal gall-bladder is removed after experiment, with normal saline flushing, is visually observed.
Physiological saline group animal gall-bladder is smooth complete, has no that mucous membrane falls off and damages;
Methyl tertiary butyl ether group mucosal erosion falls off, and there are the blutpunkte being dispersed in, no mucoid substance covering in surface;
Methyl tertiary butyl ether adds 1 bear gall powder group animal gall-bladder of embodiment smooth complete, has no that mucous membrane falls off and damages, surface has
One layer of apparent mucous substance covering;
Tri- groups of experiment group B, test group C, test group D animal gall-bladders are complete, have no that apparent mucous membrane falls off and damages, surface
Apparent mucous substance is there are no to cover, but the accidental individual blutpunktes of three groups of animals.
The above results show that bear gall powder has the remarkable result protected and improve gallbladder function, avoid methyl tertiary butyl ether pair
The damage of gall-bladder;In addition, test group A has more significant effect than it in experiment group B, test group C, tri- groups of test group D, it is special
It is not the mucous substance covering generated on gall-bladder surface to protect gall-bladder and improve the effect of gallbladder function.
The result and (2) gall-bladder pathology for dissolving the influence of gall stone dissolve stone rate to methyl tertiary butyl ether according to above-mentioned (1) are examined
The result looked into as it can be seen that bear gall powder of the present invention than it in existing bear gall powder or the bear gall powder of prior art preparation, have more
Significant dissolving gall stone or the effect for promoting dissolving gall stone, and have the effect of improving and protecting gallbladder function, this
For kind than it in the effect of existing bear gall powder or the bear gall powder of prior art preparation, producing cause may be due to passing through this
Bear gall powder obtained by inventive method has the physicochemical property different from the bear gall powder of existing bear gall powder or prior art preparation.From
It is said in this meaning, bear gall powder expection of the present invention will can be effectively used for preventing or treating gall stone disease and protection and improve courage
Capsule function.
Alanine aminotransferase (ALT), Lomefloxacin aspartate injection (AST), gamma glutamyltransferase (γ-GT or be GGT)
And AST/ALT values are the important Testing index of liver function, above-mentioned enzyme increases the damage that can reflect liver function from different perspectives
Wound.The experiment and result of above-mentioned enzyme index are described in detail below.
(3) front and back blood AST expression activitiys are tested --- to the effect of liver
The front and back blood AST expression activitiys of experiment, each group is compared with before group experiment:Nothing after the experiment of physiological saline blank control group
Difference has after the experiment of methyl tertiary butyl ether control group and significantly increases (p<0.05), (methyl tertiary butyl ether adds 1 bear gall of embodiment to test group A
Powder) indifference after experiment, it is increased significantly (p after experiment group B, test group C, test group D experiment<0.1) but not as good as methyl- tert fourth
Ether control group is notable.Concrete outcome see the table below (n=6, mean value ± s):
Group/blood AST activity | Before experiment (U/L) | Mean value value added (U/L) after experiment |
Blank control group (physiological saline) | 16.23±1.42 | 0.38 |
Control group (methyl tertiary butyl ether) | 15.51±1.76 | 3.42 |
Test group A (methyl tertiary butyl ether adds 1 bear gall powder of embodiment) | 16.64±1.57 | -0.17 |
Experiment group B (methyl tertiary butyl ether adds embodiment 13 to join 1 gained bear gall powder of embodiment) | 15.78±1.46 | 1.33 |
Test group C (methyl tertiary butyl ether adds Z26 bear gall powders) | 14.96±1.18 | 1.18 |
Test group D (methyl tertiary butyl ether adds 002# bear gall powders) | 16.84±1.95 | 1.47 |
(4) front and back blood ALT expression activitiys are tested --- to the effect of liver
The front and back blood ALT expression activitiys of experiment, each group is compared with before group experiment:Nothing after the experiment of physiological saline blank control group
Difference has after the experiment of methyl tertiary butyl ether control group and significantly increases (p<0.05), (methyl tertiary butyl ether adds 1 bear gall of embodiment to test group A
Powder) indifference after experiment, it is increased significantly (p after experiment group B, test group C, test group D experiment<0.1) but not as good as methyl- tert fourth
Ether control group is notable.Concrete outcome see the table below (n=6, mean value ± s):
(5) front and back blood GGT (gamma glutamyltransferase) expression activitiy is tested --- to the effect of liver
The front and back blood GGT expression activitiys of experiment, each group is compared with before group experiment:Nothing after the experiment of physiological saline blank control group
Difference has raising but without conspicuousness (p after the experiment of methyl tertiary butyl ether control group<0.1), (methyl tertiary butyl ether adds embodiment to test group A
1 bear gall powder) indifference after experiment, there is raising after experiment group B, test group C, test group D experiment but is compareed not as good as methyl tertiary butyl ether
Group is apparent.Concrete outcome see the table below (n=6, mean value ± s):
Group/blood GGT activity | Before experiment (U/L) | Mean value value added (U/L) after experiment |
Blank control group (physiological saline) | 9.32±1.46 | 0.07 |
Control group (methyl tertiary butyl ether) | 8.96±1.06 | 0.57 |
Test group A (methyl tertiary butyl ether adds 1 bear gall powder of embodiment) | 8.85±1.53 | 0.02 |
Experiment group B (methyl tertiary butyl ether adds embodiment 13 to join 1 gained bear gall powder of embodiment) | 9.13±1.21 | 0.31 |
Test group C (methyl tertiary butyl ether adds Z26 bear gall powders) | 9.26±1.63 | 0.28 |
Test group D (methyl tertiary butyl ether adds 002# bear gall powders) | 8.92±1.35 | 0.26 |
(6) gall-bladder liver organization inspection nearby --- to the effect of liver
Gall-bladder liver organization nearby is taken, normal saline flushing is clean, visually observes, and hepatic tissue color is ruddy, each group animal
Notable difference is not found.It observes under an optical microscope:The close no inflammation cell of physiological saline blank control group animal liver cell
Infiltration, the visible a small amount of neutrophil infiltration of methyl tertiary butyl ether control animals hepatic tissue, the close nothing of test group A animal liver cells
Inflammatory cell infiltration, experiment group B, test group C and the accidental a small amount of neutrophil infiltration of test group D each group different animals hepatic tissues
And each group most animals there are no neutrophil infiltration.
Above with regard to ALT, AST, GGT and liver organization inspection, the results showed that, bear gall powder, which has, protects and improves liver work(
The remarkable result of energy, avoids damage of the methyl tertiary butyl ether to liver;In addition, test group A than it in experiment group B, test group C, examination
Testing tri- groups of group D has more significant effect, and liver can be especially protected in terms of ALT, AST, GGT and capsule tissue and is changed
The effect of kind liver function.
The result of the result and (2) hepatic pathology inspection that are influenced on liver enzyme according to above-mentioned (3)~(6) is as it can be seen that the present invention
For bear gall powder than it in existing bear gall powder or the bear gall powder of prior art preparation, having more significantly improves and protects liver
The effect of function, it is this than it in the effect of existing bear gall powder or the bear gall powder of prior art preparation, producing cause can
Can be since gained bear gall powder has with existing bear gall powder or the bear gall powder of prior art preparation not by the method for the invention
Same physicochemical property.In this sense, bear gall powder expection of the present invention will can be effectively used for preventing or treating liver diseases example
Such as liver fibrosis and protection and improvement liver function.
Test example 5:Bear gall powder inhibits the pharmacodynamics test of mice transplanted tumor
1, test material
Drug:
Experiment is respectively with bear gall powder:1 bear gall powder of embodiment (it can be described as E1 herein), embodiment 13 are with reference to implementation
Bear gall powder (it can be described as E13c1 herein) obtained by 1 method of example, (it can be described as Z20000026 bear gall powders herein
Z26), 002# bear gall powders (it can be described as 002# herein);Bear gall powder adult daily dosage 600mg, with warm water tune before experiment
Even (40mg/ml) sets 4 DEG C of preservations during experiment.Syklofosfamid ampoule is purchased from Shanghai Hua Lian.Animal:Cleaning grade NIH mouse
Licensing SCXK (army) 2002-004;Cleaning grade C57BL/6N mouse licensings SCXK (army) 2002-003;Cleaning grade DBA/2
Mouse licensing SCXK (capital) 2000-0006.Tumor kind:Mice transplanted tumor Lewis lung cancer, P388 leukaemia, S180 sarcomas
It is purchased from institute of Materia Medica,Chinese Academy of Medical Sciences tumour room with H22 liver cancer.
2, test method
(1) Mice Bearing Lewis Lung Cancer:
Experiment shares the C57BL/6N mouse 180 of 18~22g of weight, wherein female 120, male 60.Experiment point
3 batches repeat, and mouse 60 is used in every batch of experiment, and gender is consistent.
Mouse is divided to 6 groups at random, every group 10, is set to saline control group, cyclophosphamide-a control group, bear gall powder
E1 groups, bear gall powder E13c1 groups, bear gall powder Z26 groups, bear gall powder 002# groups.
Cervical dislocation puts to death the C57BL/6N tumor-bearing mices of Lewis lung carcinoma subcutaneous inoculation growth 12d, sterile to take fresh
Tissue, cell homogenates is made with tissue grinder, and living cells content is adjusted to 2~3 × 10 with physiological saline7/ mL, is inoculated in
C57BL/6N right side of mice armpits are subcutaneous, every inoculation 0.1mL.
Dosage and method:Physiological saline, gavage, daily 0.1mL/10g body weight/days, one time a day, totally 11 times;Ring phosphorus
Amide, intraperitoneal injection, accumulated dose 80mg/kg, 20mg/kg weight/time, every other day 1 time, totally 4 times;Various bear gall powders, gavage,
200mg/kg body weight/days, one time a day, totally 11 times.
It is inoculated with the 2nd day and starts to be administered, after the last administration for 24 hours, put to death animal, weigh and tumor mass weight, be calculated as follows
Tumor-like hyperplasia:
Tumor-like hyperplasia (%)=(1- test groups knurl weight/saline control group knurl weight) × 100%.
Three batches of experiments merge statistics, the experimental data of weight and tumor weight for statistical analysis with SPSS10.0 softwares
It is indicated with x ± s, with the difference of one-way analysis of variance more each administration group and saline control group.
(2) mouse P388 leukaemia
The sterile DBA/2 mouse ascites for taking P388 leukaemia intraperitoneal inoculations growth 8d, are contained with normal saline dilution oncocyte
It measures to 1~2 × 107/ mL, it is subcutaneous to be inoculated in DBA/2 right side of mice armpits, every inoculation 0.1mL.Dosage period 11d, grouping and
Administrations, statistical method are identical as Mice Bearing Lewis Lung Cancer experiment.
(3) mouse S 180 sarcoma
The sterile mouse ascites for taking S180 sarcoma intraperitoneal inoculations growth 8d, with normal saline dilution oncocyte content to 3~4
×107/ mL, it is subcutaneous to be inoculated in armpit on the right side of experiment mice, every inoculation 0.1mL.Dosage period 11d, grouping and administrations,
Statistical method is identical as Mice Bearing Lewis Lung Cancer experiment.
(4) mouse H22 liver cancer
Take H22 liver cancer intraperitoneal inoculations growth 9d mouse ascites, with normal saline dilution to oncocyte content be 5~6 ×
107/ mL is inoculated with NIH mouse.Dosage period 11d, grouping and administrations, statistical method are identical as Mice Bearing Lewis Lung Cancer experiment.
3, test result
(1) to the histamine result of mice transplanted tumor
The growth of 4 kinds of tumor strains of positive control Cytoxan pair has apparent inhibiting effect, is 80mg/kg in accumulated dose
When, the tumor-like hyperplasia to Mice Bearing Lewis Lung Cancer, P388 leukaemia, S180 sarcomas and H22 liver cancer is respectively 82.4%,
65.7%, 69.3% and 52.5%;Bear gall powder E1, E13c1, Z26,002# distinguish the tumor-like hyperplasia of Lewis lung cancer
It is 45.7%, 25.3%, 31.6% and 22.4%;The knurl weight of bear gall powder E1, E13c1, Z26,002# to P388 leukaemia
Inhibiting rate is respectively 57.2%, 35.1%, 28.9% and 31.6%;Bear gall powder E1, E13c1, Z26,002# are to S180 meat
The tumor-like hyperplasia of tumor is respectively 44.2%, 31.2%, 38.9% and 35.4%;Bear gall powder E1, E13c1, Z26,002#
Tumor-like hyperplasia to H22 liver cancer is respectively 51.4%, 29.6%, 36.3% and 31.7%.By these results as it can be seen that bear gall powder
E1 has apparent better tumor-inhibiting action in other bear gall powders than it for kinds of tumors.This more excellent tumor killing effect may
It is since specific preparation method of the invention assigns bear gall powder excellent properties.
(2) to the toxic side effect of tumor-bearing mice
In the therapeutic test of mice transplanted tumor, various bear gall powders do not result in mouse weight growth and obviously slow down,
Have no animal dead and other adverse reactions, this shows that bear gall powder of the present invention has excellent safety.
Test example 6:Bear gall powder platelet aggregation inhibitory activity is tested
Rat divided to 6 groups at random, every group 6, is set to control group (0.5%CMC-Na), (dosage is aspirin
25mg/kg, aspirin enteric coated tablet 2 go film-coating to smash to be dissolved in 0.5%CMC-Na, be made into the solution of 5mg/mL),
Bear gall powder E1 groups, bear gall powder E13c1 groups, bear gall powder Z26 groups, bear gall powder 002# groups, each bear gall powder group dosage is 500mg/kg.
Each group gastric infusion.After 120min is administered, eye socket takes 4mL blood, is placed in the test tube of 3.8% sodium citrate containing 0.4mL,
After mixing platelet rich plasma (PRP) and platelet poor plasma (PPP) are prepared by different centrifugal speeds respectively.1000r/min
Upper plasma, which is drawn, after centrifugation 10min obtains PRP;PPP is obtained by upper plasma is drawn after residual blood 2500r/min centrifugations 10min.
It takes the PPP of 250 μ L to be added in PAPE-I types platelet aggregation/blood clotting analyzer test cup, is put into instrument connection and calibrates.Take 250 μ
The PRP of L is added in test cup, and 1min is preheated in 37 DEG C of pre-temperature slots, and 2 μ g/mL derivants collagen, 25 μ L are added immediately, measures blood
Platelet maximum aggregation rate.3 channels are measured every time, calculate average maximum aggregation rate.With SPSS16.0 statistical analysis softwares, t
It examines and carries out data processing, as a result:The blood platelet maximum aggregation inhibiting rate of aspirin group is the blood of 36.6%, bear gall powder E1 groups
Platelet maximum assemble inhibiting rate be 34.8%, bear gall powder E13c1 groups, bear gall powder Z26 groups, bear gall powder 002# group threes blood platelet
Maximum aggregation inhibiting rate is respectively 21.6%, 24.1%, 19.7%.The result shows that bear gall powder E1 is than its resisting in other bear gall powders
Platelet aggregation activity is stronger.
Test example 7:Bear gall powder inhibition thrombosis is tested
This experiment is by studying the antithrombotic effect of bear gall powder, to clinical indication (such as thrombus to open up new
Formative disease) experimental basis is provided.
Kunming mouse 90, half male and half female are randomly divided into 6 groups.Physiological saline (control group, 0.2mL/g), bear are given respectively
Courage powder E1 groups, bear gall powder E13c1 groups, bear gall powder Z26 groups, bear gall powder 002# groups, warfarin (10mg/kg) gavage;Each bear gall powder
Group dosage is 500mg/kg.Above-mentioned dosage 1 time a day, for three days on end.At 1h after the last administration (No Food or Drink in 1h), with hair
Thin glass tube is inserted into mouse intraocular corner of the eyes ball rear vein beard, and deep about 4~5mm is gently rotated and retracted again.Autoblood, which flows into pipe, to be started
Timing, blood take out glass capillary after filling and lie against desktop, fractures both ends glass capillary about 0.5cm every 30s, and delay
Slowly it pulls open to the left and right, whether the observation place of fractureing has blood clotting silk, until blood clotting silk occurs, when required time is thrombosis
Between, it is averaged as thrombosis time (T) [Beijing Qi Chen herbal pharmacologies research methodology [M] of the mouse:The people defend
Raw publishing house, 1993:484-485].
As a result:The mean value of control group thrombosis time is 56.4 seconds, and the mean value of warfarin thrombosis time is 262.5
Second, the mean value of bear gall powder E1 group thrombosis times is 183.2 seconds, bear gall powder E13c1 groups, bear gall powder Z26 groups, bear gall powder 002#
The mean value of the thrombosis time of group three is respectively 91.3 seconds, 114.5 seconds, 89.8 seconds.The result shows that bear gall powder E1 than it in
The inhibition thrombosis activity of other bear gall powders is stronger.
Thrombosis disease seriously endangers the life and health of the mankind, and incidence is in first of various diseases.Various originals
An important factor for whole body caused by or regional flow are slowly thrombosis such as high microsteping proteinemia, hyperlipidemia, takes off
Hyperviscosity syndrome and dyshaemia caused by water, erythremia etc..Thrombosis Mechanism:1. erythrocyte aggregation is agglomerating, shape
At red thrombus;2. blood platelet sticks and assembles, discharges and react with endothelium;3. blood vessel endothelium injury starts coagulation process
[leaf appoints the Beijing high clinical practice [M]:People's Health Publisher, 2002:689-691].Clinically commonly use anticoagulant or fibre
Fibrillarin dissolves medicine to treat thrombosis disease or carry out antithrombotic, such as heparin, Coumarins, streptokinase.
In recent years, Chinese medicine thrombosis disease is increasingly subject to the attention of domestic and foreign scholars.The display of this experimental result, bear gall powder
The bear gall powder that especially the method for the present invention is prepared has excellent inhibition thrombosis activity.
Test example 8:The experiment that bear gall powder improves immunity of organisms, improves muscle power, alleviates organism fatigue
1, experimental animal and grouping
Male and healthy Wistar rats 40 and the male hyperlipidemia Wistar rats 40 for taking weight (200 ± 10) g,
Before experiment, rat is placed on laboratory rearing one week to make rat adapt to environment, it, will be healthy then according to randomized blocks
Wistar rats (big group of A) are divided into A bear gall powder E1 groups, A bear gall powder E13c1 groups, A bear gall powder Z26 groups, A bear gall powder 002# groups, A
Control group totally five groups, by male hyperlipidemia Wistar rats (big group of B) be divided into B bear gall powder E1 groups, B bear gall powder E13c1 groups,
B bear gall powder Z26 groups, B bear gall powder 002# groups, B control groups totally five groups, every group of 8 animals of each group.
2, administration and measurement
A control groups and B control rats use normal rats feed, and all four groups are using normal rats to big group of A
10g or less bear gall powders are added respectively in every 100 grams of feeds on the basis of feed:Bear gall powder E1, bear gall powder E13c1, bear gall powder
All four groups are mixed on the basis of using normal rats feed in every 100 grams of feeds respectively by Z26, big group of bear gall powder 002#, B
Add 10g or less bear gall powders:Bear gall powder E1, bear gall powder E13c1, bear gall powder Z26, bear gall powder 002#.
The rat of 10 groups is continuously raised 10 weeks.After raising, by Rat Fast 12h, sterile taking-up spleen separation
Obtain splenic lymphocytes.
Measure lymphoproliferation activit [Morten BH, Svend EN, Kurt B.Re-examination and
further development of a precise and rapid dye method for measuring cell
Growth/cell kill [J] .J of Immunological Methods, 1989,119 (2):203-210;Lin Zhongning, Dong
Win jade tablet, Dong Shuyun waits the Study of Methodology of .MTT methods detection T the proliferative function of lymphocyte to be examined with application [J] China Healths miscellaneous
Will, 2000,10 (1):8-10].The spleen aseptically taken out is pulverized to and is fabricated to individual cells suspension, by cell number
It is adjusted to 4 × 109A/L.96 hole flat undersides are added in cell suspension after adjustment to cultivate, 200 μ L are added per hole, add altogether
Enter 8 holes, 8 holes are divided into positive hole and negative hole, wherein 4 hole of positive hole, wherein the knife of 25 a concentration of 50mg/L of μ L is added per hole
Legumin (ConA), 4 hole of negative hole are then added without ConA, and the flat-bottomed plates after adding well are placed in CO2 incubators, at a temperature of 37 DEG C
Cultivate 48h.The 4h before culture terminates the MTT10 of 10 a concentration of 5g/L of μ L is added in each hole of 96 hole flat-bottomed plates,
Culture terminates preceding 2h, and 100 μ L buffer solutions are added in each hole for measuring optical density, start to measure optical density after adding.Its
In, the proliferative capacity of T lymphocytes is reflected with stimulus index (Stimulation Index, SI).The sun of SI=ConA stimulations
The average optical density value for the negative hole that the average optical density value/ConA in property hole is not stimulated.
Proleulzin and interleukin-4 (IL-2, IL-4) measure.Lymphocyte suspension is separately added into the flat culture in 24 holes
The ConA of 100 a concentration of 50mg/L of μ L is then added to every hole, is put into CO2 incubators and cultivates per hole 1mL again for plate, temperature 37
DEG C, incubation time 48h.After culture, after the lymphocyte supernatant in culture plate is aseptically drawn, then distinguish
It is fitted into the EP pipes of 0.5mL, is preserved in the refrigerator of -20 DEG C of merging, be used for the detection of IL-2 and IL-4 contents.It is examined using ELISA
The content of test agent box detection detection IL-2 and IL-4.
3, data record and statistical analysis
The activity condition of each group rat is observed during raising and death condition and is recorded;To each group's rat lymphocyte
Proliferative capacity stimulus index and proleulzin, 4 results recorded.This research institute obtains data and is all made of SPSS20.0
It is handled, component compares to be examined using t, is indicated with (x ± s), with P<0.05 has differences for statistics.
In addition the energy and vigor of each group animal are observed during experiment and after the test, and are recorded each when off-test
The energy vigor score (being indicated with the mean value of every group of 8 animals) of group animal, the energy vigor score observe by the naked eye, 10 points
Highest and indicate energy vigor highest, 0 point it is minimum and indicate that energy vigor is minimum.
The energy and vigor of animal can reflect the muscle power height and degree of fatigue of animal, the higher expression of energy vigor score
Animal muscle power is high and abundant, endurance, the low and weak, fatiguability of lower the expressions animal muscle power of energy vigor score.
4, result
During raising, each group rat does not occur death, and make discovery from observation A bear gall powder E1 groups and B bear gall powders
The rat vigor and energy of E1 groups are better than other rats, and the energy of B control rats and vigor are worst.
Mice spleen lymphocytes proliferation activity and interleukin content balance see the table below.
Note:In big group of A, there were significant differences for SI indexes, IL-2 values and IL-4 values compared with A control groups for bear gall powder E1 groups
(P<0.05), test group is obviously improved than three parameters of control group, and in addition three kinds of bear gall powders SI compared with A control groups refers to
Number, IL-2 values and the variant (P of IL-4 values<0.1) but do not have conspicuousness;In big group of B, bear gall powder E1 groups and B control group phases
Than SI index, IL-2 values and IL-4 values there were significant differences (P<0.05), test group is obviously carried than three parameters of control group
Height, in addition three kinds of bear gall powders SI indexes, IL-2 values and variant (P of IL-4 values compared with B control groups<0.1) but do not have significantly
Property.
The above result shows that bear gall powder of the present invention can significantly increase the immunity of healthy rat and hyperlipemia rat,
And the energy and vigor of animal body are improved, it is worthy of popularization.
Above disclosed is only presently preferred embodiments of the present invention, cannot limit the right of the present invention with this certainly
Range, therefore according to equivalent variations made by scope of the present invention patent, be still within the scope of the present invention.
Claims (10)
1. a kind of bear gall powder is the burnished gold or bright-yellow powder of hyaloid gloss, glassy yellow fluorescence is shown at 365nm,
For example≤1%, and its solubility in water is more than 10g/100ml to hydroscopicity≤2%, and the solubility in 90% ethyl alcohol is big
In 1.2g/100ml, the pH value of 5% aqueous solution is in 6.0~7.5 ranges.
2. bear gall powder according to claim 1,:
Solubility in water is 13.6~14.2g/100ml;
Solubility in 90% ethyl alcohol is 1.25~2g/100ml;
The pH value of 5% aqueous solution is in 6.0~7.0 ranges;
Moisture is 0.4~1%.
3. bear gall powder according to claim 1 is prepared by a method comprising the following steps to obtain:
(1) the bear gall juice by the acquisition of no Tube Drain method filters, and obtains filtrate, is diluted with water, obtain bear bile fluid, at 2~6 DEG C of temperature
12 hours are stood, then carries out subsequent operation;
(2) it is filled with 20~30min of nitrogen into bear bile fluid obtained by step (1), is then filled with 10~20min of carbon dioxide;
(3) bear bile fluid handled through aforesaid operations is inclined in material disc, is immediately placed on and is refrigerated to -45 DEG C of freezings below in advance
In drying machine, freeze 2~5 hours;Then -25~-20 DEG C are warming up to 0.25~0.35 DEG C of speed per minute, keep 1~
1.5 hour;Then it is warming up to -5~0 DEG C with 0.25~0.35 DEG C of speed per minute, is kept for 2~3 hours;Then with per minute
0.25~0.35 DEG C of speed is warming up to 10~15 DEG C, is kept for 4~5 hours;Then with 0.25~0.35 DEG C of speed per minute
30~35 DEG C are warming up to, is kept for 2~3 hours, freeze-drying program is terminated;
(4) by the dried lump in material disc be ground into can by the fine powder that 50 mesh sieve to get.
4. bear gall powder according to claim 1, wherein:
In step (1) plus the water of 0.5~1 times of amount dilutes;
When inflatable body it is inflated in a manner of bubbling in step (2);
Step is filled with 25~30min of nitrogen in (2);
Step is filled with 15~20min of carbon dioxide in (2).
5. bear gall powder according to claim 1, wherein:
Bear bile fluid is inclined in material disc in step (3), and liquid layer depth is less than 5cm;
Bear bile fluid is inclined in material disc in step (3), and liquid layer depth is less than 2.5cm;
Bear bile fluid is inclined in material disc in step (3), and liquid layer depth is 1.5~2.5cm.
6. bear gall powder according to claim 1, wherein:
In step (3), so that material disc is immediately placed on and be refrigerated in -55~-45 DEG C of freeze drier in advance;
In step (3), so that material disc is immediately placed on and be refrigerated in -50~-45 DEG C of freeze drier in advance.
7. bear gall powder according to claim 1, wherein:
In step (3), so that material disc is immediately placed on and be refrigerated in -50~-45 DEG C of freeze drier in advance, freezing 3~4 is small
When;Then -25~-20 DEG C are warming up to 0.3 DEG C of speed per minute, are kept for 1~1.5 hour;Then with 0.3 DEG C per minute
Speed is warming up to -5~0 DEG C, is kept for 2~3 hours;Then 10~15 DEG C are warming up to 0.3 DEG C of speed per minute, keep 4~5
Hour;Then 30~35 DEG C are warming up to 0.3 DEG C of speed per minute, are kept for 2~3 hours, terminate freeze-drying program.
8. the method for preparing bear gall powder, includes the following steps:
(1) the bear gall juice by the acquisition of no Tube Drain method filters, and obtains filtrate, is diluted with water, obtain bear bile fluid, at 2~6 DEG C of temperature
12 hours are stood, then carries out subsequent operation;
(2) it is filled with 20~30min of nitrogen into bear bile fluid obtained by step (1), is then filled with 10~20min of carbon dioxide;
(3) bear bile fluid handled through aforesaid operations is inclined in material disc, is immediately placed on and is refrigerated to -45 DEG C of freezings below in advance
In drying machine, freeze 2~5 hours;Then -25~-20 DEG C are warming up to 0.25~0.35 DEG C of speed per minute, keep 1~
1.5 hour;Then it is warming up to -5~0 DEG C with 0.25~0.35 DEG C of speed per minute, is kept for 2~3 hours;Then with per minute
0.25~0.35 DEG C of speed is warming up to 10~15 DEG C, is kept for 4~5 hours;Then with 0.25~0.35 DEG C of speed per minute
30~35 DEG C are warming up to, is kept for 2~3 hours, freeze-drying program is terminated;
(4) by the dried lump in material disc be ground into can by the fine powder that 50 mesh sieve to get.
9. the bear gall powder of any one of claim 1-7 is being prepared for improving immunity of organisms, improving muscle power, alleviation organism fatigue
Product in purposes.
10. purposes according to claim 9, wherein the product is drug, health food.
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CN108653332A (en) * | 2018-07-14 | 2018-10-16 | 齐光伟 | Inhibition thrombosis and platelet aggregation and the bear gall powder and purposes for preventing cerebral ischemia |
CN108785328A (en) * | 2018-07-14 | 2018-11-13 | 齐光伟 | Prevent or treat method and the bear gall powder used of entity tumor and hematological system tumor |
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CN108578433A (en) * | 2018-07-14 | 2018-09-28 | 齐光伟 | For preventing or treating gall stone cholecystitis and improve the bear gall powder of gallbladder function |
CN108578434A (en) * | 2018-07-14 | 2018-09-28 | 齐光伟 | Prevent or treat method and the bear gall powder used of eye conjunctivitis and eyelid herpes zoster |
CN108635375A (en) * | 2018-07-14 | 2018-10-12 | 齐光伟 | Prevent or treats hepatopathy and liver fibrosis and improve the bear gall powder and purposes of liver function |
CN108653332A (en) * | 2018-07-14 | 2018-10-16 | 齐光伟 | Inhibition thrombosis and platelet aggregation and the bear gall powder and purposes for preventing cerebral ischemia |
CN108785328A (en) * | 2018-07-14 | 2018-11-13 | 齐光伟 | Prevent or treat method and the bear gall powder used of entity tumor and hematological system tumor |
WO2023088463A1 (en) * | 2021-11-22 | 2023-05-25 | Shenzhen Hightide Biopharmaceutical Ltd. | Compositions and methods of berberine ursodeoxycholate for modulating gut microbiome compositions |
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