KR101662887B1 - Composition Comprising Actinidia arguta shoot extract for prevention or treatment of nonalcoholic fatty liver disease - Google Patents

Abstract

The present invention relates to a composition for the prevention or treatment of nonalcoholic fatty liver disease comprising an extract of Quercus grandiflorum as an active ingredient. The net extract of the present invention has the effect of reducing the blood glucose and serum insulin in the body and reducing the lipid content and triglyceride content of the liver due to the ingestion of high fat and high sugar and is effective in reducing the inflammatory cytokine TNF- , MCP-1 and IL-6, and has an anti-inflammatory activity. Thus, the extract of the present invention can be effectively used for prevention or treatment of non-alcoholic fatty liver disease. In particular, since the extract of this invention is a natural substance, it corresponds to a medicament widely used as a herbal medicine. Therefore, the composition of the present invention containing it as an effective ingredient has a safety advantage even in long-term use.

Description

Technical Field [0001] The present invention relates to a composition for preventing or treating nonalcoholic fatty liver disease,

The present invention relates to a composition for preventing or treating a non-alcoholic fatty liver comprising an extract of Quercus grandiflorum as an active ingredient.

Fatty liver disease, also called fatty liver disease, is a disease caused by abnormal accumulation of fat (triglycerides, etc.) in hepatocytes, resulting in hepatic damage. It is known that the initial condition of fatty liver disease is a simple fatty liver that recognizes only fat deposition in hepatocytes, and then the condition progresses to fatty liver (including liver fibrosis), and also cirrhosis or hepatocellular carcinoma. In general, the causes of fatty deposits in the liver include alcohol consumption, obesity, diabetes, lipid metabolism disorders, drugs (steroids, tetracyclines, etc.), Cushing's syndrome, poisoning .

The causes of fatty liver disease are largely divided into alcohol and non-alcoholic, and liver disease caused by the former is called alcoholic fatty liver disease (also called alcoholic hepatopathy) and the latter is caused by non-alcoholic fatty liver disease nonalcoholic fatty liver disease (NAFLD).

Alcoholic fatty liver disease progresses from the initial simple fatty liver to fatty liver and cirrhosis. Non-alcoholic fatty liver disease has been thought to remain in simple fatty liver and the condition does not progress. However, recently, it has been found that non-alcoholic fatty liver disease also progresses from simple fatty liver to fatty liver or liver cirrhosis.

Nonalcoholic steatohepatitis (NAFLD) is a non-alcoholic steatohepatitis (NAFLD) that develops in the liver and is accompanied by hepatic inflammation (hepatocellular inflammation) (Ludwig J et al., Mayo Clin Proc 1980, 55 (7): 434-438), which refers to a wide range of diseases including liver fibrosis and cirrhosis, .

Non-alcoholic fatty liver disease is a disease characterized by fatty changes (steatosis) and lobular hepatitis (steatohepatitis), which are hallmarks of alcoholic hepatitis in liver biopsy despite the absence of alcohol- ) And so on. The pathologic findings of the liver are diverse spectrum from non-alcoholic fatty liver (NAFL) to non-alcoholic steatohepatitis (NASH), fibrosis associated with fatty liver, liver cirrhosis, It is used to mean all inclusive.

Nonalcoholic fatty liver disease is divided into primary and secondary, depending on the cause. Primary is caused by hyperlipidemia, diabetes, or obesity, which is characteristic of metabolic syndrome. Secondary is caused by nutritional causes (rapid weight loss, starvation, ), Various drugs (glucocorticoids, estrogens, tamoxifen, methotrexate, zidovudine, amiodarone, tetracycline, didanosine, cocaine, diltiazem, perhexiline), toxic substances (poisonous bacteria, bacterial toxins), metabolic causes (lipodystrophy, dysbetalipopoteinemia, Weber-Christian syndrome, It is known to be caused by inflammatory bowel syndrome (HIV infection) and other factors (Reye's syndrome) and acute fatty liver of pregnancy. It is known that the incidence of nonalcoholic fatty liver disease related to diabetes and obesity, which is an important factor of metabolic syndrome, is almost 50% in diabetic patients, 76% in obese patients, and almost in obese diabetic patients. The incidence of hepatitis was 18 to 36% in patients with diabetes and obesity with elevated levels of alanine aminotransferase (ALT). The hepatitis was diagnosed by insulin resistance Is known to occur. The rate of progression to liver cirrhosis in patients with diabetes mellitus, obesity, and hyperlipidemia varies according to the duration of the disease, and the proportion of patients progressing from cirrhosis to liver cirrhosis within 3 to 11 years is 4 to 26 %, And it is reported that the mortality rate is higher than the general population.

In addition, liver accumulation of triglycerides directly associated with nonalcoholic fatty liver disease and hepatocellular damage resulting therefrom are caused by changes in systemic factors such as local factors and insulin resistance, Synthesis and release / oxidation imbalance. Gastroenterology 2001, 121 (3) (2001), that fatty acids are introduced into the liver in excess of the fatty acids that can be oxidized by the mitochondria of hepatocyte due to insulin resistance caused by hyperinsulinemia (Reid AE, : 710-723) are known to accumulate triglycerides. In addition, insulin resistance causes lipogenic transcription factors

The expression of peroxisome proliferator-activated receptor-gamma (PPAR-gamma) and SREBP-1c (upregulation of sterol regulatory element binding protein-1c) Accumulation of triglycerides may also be present.

There is no established treatment for this nonalcoholic fatty liver disease because it is associated with diverse factors such as diabetes, obesity, coronary artery disease, and sitting habits. Obesity is an important target in the treatment of nonalcoholic fatty liver disease, and is associated with factors associated with insulin resistance, the amount of fatty acids entering the liver, and inflammatory or fibrotic adipokines As well. Alterine alanine aminotransferase (ALT) levels and triglyceride content in the liver may be reduced by dietary control and exercise-induced weight loss, but improvements in weight loss in patients with necrotizing inflammation or liver fibrosis Little is known.

As described above, a sufficient therapeutic method has not been established for non-alcoholic fatty liver disease, and development of a therapeutic drug having high effectiveness against non-alcoholic fatty liver disease has been desired.

Korean Patent Application No. 2011-0016416

Accordingly, the inventors of the present invention have completed the present invention by confirming that the extract of Quercus grandiflorus can improve and treat the symptoms of non-alcoholic fatty liver disease while studying natural products as a therapeutic agent for treating non-alcoholic fatty liver disease.

It is therefore an object of the present invention to provide a process for the production of Actinidia The present invention also provides a pharmaceutical composition for preventing or treating non-alcoholic fatty liver disease, which comprises an extract of arguta shoot as an active ingredient.

A further object of the present invention is to provide a method for the production of Actinidia The present invention provides a health functional food for preventing or ameliorating a non-alcoholic fatty liver disease comprising an extract of arguta shoot as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating non-alcoholic fatty liver disease comprising Actinidia arguta shoot extract as an active ingredient.

In one embodiment of the present invention, the extract may be obtained by extracting with water or an organic solvent.

In one embodiment of the present invention, the organic solvent may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane and cyclohexane .

In one embodiment of the present invention, the net extract may be contained in an amount of 0.1 to 10% by weight based on the total weight of the composition.

In one embodiment of the present invention, the net extract of the present invention may be effective in reducing blood glucose and serum insulin in the body and reducing lipid content and triglyceride content of the liver due to ingestion of high fat and high sugar have.

In one embodiment of the present invention, the extract may be one having anti-inflammatory activity by inhibiting the production of inflammatory cytokines TNF-a, MCP-1 and IL-6.

In one embodiment of the present invention, the non-alcoholic fatty liver disease may be selected from the group consisting of non-alcoholic simple fatty liver, non-alcoholic fatty liver and non-alcoholic cirrhosis.

The present invention also relates to a method for the treatment of Actinidia The present invention provides a health functional food for preventing or ameliorating a non-alcoholic fatty liver disease comprising an extract of arguta shoot as an active ingredient.

In one embodiment of the present invention, the extract may be obtained by extracting with water or an organic solvent.

In one embodiment of the present invention, the organic solvent may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane and cyclohexane .

In one embodiment of the present invention, the net extract may be contained in an amount of 0.1 to 10% by weight based on the total weight of the composition.

In one embodiment of the present invention, the net extract of the present invention may be effective in reducing blood glucose and serum insulin in the body and reducing lipid content and triglyceride content of the liver due to ingestion of high fat and high sugar have.

In one embodiment of the present invention, the extract may be one having anti-inflammatory activity by inhibiting the production of inflammatory cytokines TNF-a, MCP-1 and IL-6.

In one embodiment of the present invention, the non-alcoholic fatty liver disease can be selected from the group consisting of non-alcoholic simple fatty liver, non-alcoholic fatty liver and non-alcoholic cirrhosis.

The present invention relates to a composition for the prevention or treatment of nonalcoholic fatty liver disease comprising an extract of Quercus grandiflorum as an active ingredient. The net extract of the present invention has the effect of reducing the blood glucose and serum insulin in the body and reducing the lipid content and triglyceride content of the liver due to the ingestion of high fat and high sugar and is effective in reducing the inflammatory cytokine TNF- , MCP-1 and IL-6, and has an anti-inflammatory activity. Thus, the extract of the present invention can be effectively used for prevention or treatment of non-alcoholic fatty liver disease. In particular, since the extract of this invention is a natural substance, it corresponds to a medicament widely used as a herbal medicine. Therefore, the composition of the present invention containing it as an effective ingredient has a safety advantage even in long-term use.

FIG. 1 is a graph showing the results of analysis of the effect of controlling the blood glucose level of the extract of P. multiflorum of the present invention.
FIG. 2 is a graph showing the results of analysis of the effect of the present invention on the reduction of serum insulin by the high-fat diet and high sugar intake.
FIG. 3 is a graph showing the results of HOMA-IR measurement of the activity of improving the insulin resistance of the extract of the present invention.
FIG. 4 is a graph showing the results of analysis of the activity of reducing the lipid content in the liver tissues, which was increased by high-fat diet and high sugar intake, according to the present invention.
FIG. 5 is a graph showing the results of analysis of the activity of reducing the triglyceride content in liver tissues, which was increased by high-fat diet and high sugar intake, according to the present invention.
FIG. 6 is a graph showing the anti-inflammatory activity of the extract of P. multinense according to the present invention and showing the activity of reducing the production of TNF-a in serum.
FIG. 7 is a graph showing the results of analysis of the activity of reducing the production of MCP-1 in serum by analyzing the anti-inflammatory activity of the extract of P. multiflorum of the present invention.
FIG. 8 is an analysis of the anti-inflammatory activity of the extract of P. multiflorum of the present invention, and shows the results of analysis of the activity of reducing the production of IL-6 in serum.

The present invention relates to a novel use of a net extract of Dulone, and is characterized by providing a composition for preventing or treating a non-alcoholic fatty liver disease comprising an extract of Dulone extract as an active ingredient.

In the present invention, Actinidia arguta shoot, which is an effective ingredient for improving and treating the non-alcoholic fatty liver disease, is collected mainly in spring, cooked in a herb, or prepared as an acorn and stored for a long time. It has been studied for contents that have anti-obesity effect and antioxidant effect, but overall research on the order of Dalae has not been done yet. to be.

The present invention has the effect of reducing the increase in blood glucose and serum insulin in the body due to the ingestion of high fat and high sugar and reducing the lipid content and triglyceride content of liver tissue, Inflammatory cytokines TNF-α, MCP-1 and IL-6, and thus it is possible to prevent or treat non-alcoholic fatty liver disease.

Accordingly, the present invention can provide a composition capable of preventing or treating non-alcoholic fatty liver disease by containing the extract of Porphyra yezoensis as an active ingredient.

The extract of the genus Dulone according to the present invention can be obtained by extracting and isolating from nature using an extraction and separation method known in the art, and the 'extract' defined in the present invention can be obtained by extracting And includes, for example, crude extract, polar solvent-soluble extract, or non-polar solvent-soluble extract of Alaska pollack.

As an appropriate solvent for extracting the extract from the above-mentioned extracts, any organic solvent which is pharmaceutically acceptable may be used, and water or an organic solvent may be used, and for example, purified water, methanol alcohol having 1 to 4 carbon atoms, acetone, ether, benzene, chloroform (including methanol, ethanol, propanol, isopropanol, and butanol) various solvents such as chloroform, ethyl acetate, methylene chloride, hexane and cyclohexane may be used alone or in combination. More preferably, ethanol (alcohol) can be used.

As the extraction method, any one of the methods such as hot water extraction method, cold extraction method, reflux cooling extraction method, solvent extraction method, steam distillation method, ultrasonic extraction method, elution method and compression method can be selected and used. In addition, the desired extract may be further subjected to a conventional fractionation process or may be purified using a conventional purification method. There is no limitation to the method for producing the extract of the present invention, and any known method can be used.

For example, the net extract of the present invention may be prepared in powder form by an additional process such as vacuum distillation or freeze drying or spray drying, have. Further, the primary extract can be further purified by using various chromatographies such as silica gel column chromatography, thin layer chromatography, high performance liquid chromatography and the like, You can get it.

Therefore, in the present invention, the net extracts of Daruya comprise all the extracts, fractions and tablets obtained in each step of extraction, fractionation or purification, their diluted solutions, concentrates or dried products.

The method for preparing the extract of Dulone extract according to an embodiment of the present invention will be described in more detail as follows.

In the present invention, after drying and pulverizing the plant, the plant is treated with a lower alcohol having 1 to 4 carbon atoms, preferably about 5 to 15 times the weight of the raw sample, preferably 20 to 25 ° C for 3 to 15 hours Lt; / RTI >

The ethanol extract obtained from the above-described method can be separated from the extract and the residue by filtration under reduced pressure to obtain an extract of A. napus.

The composition of the present invention comprising the above extract of Phellodendriola vera as an active ingredient may be a pharmaceutical composition or a food composition.

The pharmaceutical composition of the present invention can be prepared by using pharmaceutically acceptable and physiologically acceptable adjuvants in addition to the above-mentioned active ingredients. Examples of the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, A lubricant or a flavoring agent can be used.

The pharmaceutical composition may be formulated into a pharmaceutical composition containing at least one pharmaceutically acceptable carrier in addition to the above-described active ingredients for administration.

The pharmaceutical composition may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, liquids, syrups, juices, suspensions, emulsions, drops or injectable solutions. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural and synthetic gums such as starch, gelatin, glucose or beta-lactose, natural sugars such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum and the like. Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include those suitable for sterilization and in vivo such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

In one embodiment of the present invention, the net extract of the present invention may be contained in an amount of 0.1 to 10% by weight based on the total weight of the composition.

The composition of the present invention may also be a food composition. The food composition may contain various flavors or natural carbohydrates as an additional ingredient, such as a conventional food composition, in addition to containing an effective ingredient, a nettle extract.

Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. The above-described flavors can be advantageously used as natural flavorings (tau martin), stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.).

The food composition of the present invention can be formulated in the same manner as the above pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, meat, chocolates, foods, confectionery, pizza, ram noodles, other noodles, gums, candy, ice cream, alcoholic beverages, vitamin complexes, .

In addition, the food composition may further contain flavorings such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and aging agents (cheese, chocolate, etc.), pectic acid, Salts of alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, the food composition of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks.

The net extract of Dulone, an active ingredient of the present invention, is a natural substance and has almost no toxicity and side effects. Therefore, it can be safely used for long-term use for the prevention or treatment of non-alcoholic fatty liver disease.

The present invention also provides a health functional food for improving liver function and improving non-alcoholic fatty liver disease, which comprises net extract of Dulone, as an active ingredient.

The health functional food of the present invention can be prepared and processed in the form of tablets, capsules, powders, granules, liquids, and circles for the purpose of improving liver function, specifically improving non-alcoholic fatty liver disease.

In the present invention, the term "health functional food" refers to foods manufactured and processed using raw materials or ingredients having useful functions in accordance with Law No. 6727 on Health Functional Foods, Or to obtain a beneficial effect in health use such as physiological action.

The health functional foods of the present invention may contain conventional food additives and, unless otherwise specified, whether or not they are suitable as food additives are classified according to the General Rules for Food Additives approved by the Food and Drug Administration, Standards and standards.

Examples of the items listed in the above-mentioned 'food additives' include chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; Natural additives such as persimmon extract, licorice extract, crystalline cellulose, high color pigment and guar gum; L-glutamic acid sodium preparations, noodle-added alkalis, preservative preparations, tar coloring preparations and the like.

For example, the health functional food in the form of tablets may be prepared by granulating a mixture obtained by mixing the active ingredient of the present invention with an excipient, a binder, a disintegrant and other additives in a usual manner, and then adding a lubricant Or the mixture can be directly compression molded. In addition, the health functional food of the tablet form may contain a mating agent or the like if necessary.

The hard capsule of the capsule-type health functional food may be prepared by filling a normal hard capsule with a mixture of the net extract of the present invention, which is an effective ingredient of the present invention, with an additive such as an excipient, etc. The soft capsule may contain, And filling the mixture with a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a coloring agent, a preservative and the like, if necessary.

The ring-shaped health functional food can be prepared by molding a mixture of the extract of Quercus grandiflorum, an effective ingredient of the present invention, excipient, binder, disintegrant, etc., by a conventionally known method and, if necessary, Or it may be coated with a material such as starch, talc.

The granular health functional food may be prepared by granulating a mixture of the net extract of the present invention and excipients, binders, disintegrators, etc. into granules by a known method, and if necessary, adding a flavoring agent, And the like.

As shown in the following examples, the health functional food comprising the extract of the present invention as the active ingredient of the present invention can significantly reduce the insulin reduction in the serum, the lipid content of the liver tissue and the triglyceride content, , Especially for non-alcoholic fatty liver improvement.

The health functional food may be a beverage, a meat, a chocolate, a food, a confectionery, a pizza, a ramen, a noodle, a gum, a candy, an ice cream, an alcoholic beverage, a vitamin complex and a health supplement food.

Further, the present invention provides the use of a composition comprising as an active ingredient, a herbal extract for preventing or treating nonalcoholic fatty liver disease or a food for producing a food. The composition of the present invention comprising the above-mentioned extract of nettle fruit extract as an active ingredient can be used for the production of medicines or foods for the prevention or treatment of non-alcoholic fatty liver disease.

The present invention also provides a method for the prevention or treatment of nonalcoholic fatty liver disease comprising administering to a mammal a nettle extract.

As used herein, the term " mammal " refers to a mammal that is the subject of treatment, observation, or experimentation, preferably a human.

As used herein, the term " therapeutically effective amount " refers to the amount of active ingredient or pharmaceutical composition that elicits a biological or medical response in a tissue system, animal or human, as contemplated by a researcher, veterinarian, The amount that induces the relief of the symptoms of the disease or disorder being treated. It will be apparent to those skilled in the art that the therapeutically effective dose and the number of administrations of the active ingredient of the present invention will vary depending on the desired effect. Thus, the optimal dosage to be administered can be readily determined by those skilled in the art and will vary with the nature of the disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation, and the age, The age, body weight, sex, diet, time of administration, route of administration and fraction of the composition, duration of treatment, concurrent medication, and the like. In the treatment method of the present invention, in the case of an adult, it is preferable to administer the herbal extract of the present invention at a dose of 1 mg / kg to 250 mg / kg once or several times a day.

In the therapeutic method of the present invention, the composition comprising the extract of the present invention as an active ingredient may be administered orally, rectally, intravenously, intraarterially, intraperitoneally, intramuscularly, intramuscularly, transdermally, topically, And can be administered in a conventional manner.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are for further illustrating the present invention, and the scope of the present invention is not limited to these examples.

< Example  1>

Dalae  Preparation of extract

Distilled water corresponding to a weight of 20 times was added to commercially available Dulaya seeds and then extracted at 90 ^° C for 8 hours. The extracts were lyophilized at 80 ^° C in a vacuum rotary evaporator and lyophilized. The lyophilized powder was extracted with 70% ethanol (10 times (w / v)) for 12 hours, 6 hours and 3 hours at room temperature The extracts were sequentially extracted and then filtered to separate the extract and the residue. The filtrate was concentrated under reduced pressure at 50 ° C using a rotary evaporator (EYELA FDU-1100, Japan) to prepare a net extract of 13.2%.

< Example  2>

Weight and Dietary intake  Measure

① Experimental animals and experiment design

Five - week - old male C57BL / 6J mice were purchased from Hyochang Science and were adapted to a weekly diet (Purina rodent chow 5057, Purina). They were divided into three groups according to their body weight. The temperature of the breeding room was maintained at room temperature of 24 ± 5 ° C, humidity was maintained at 55 ± 5%, light intensity was switched on and off at intervals of 12 hours, and animal body weight and dietary intake were measured twice a week. The composition of the experimental diet was as follows. The control group was fed with a diet containing 65% corn starch and 5% corn oil, and 27% sugar, 3% corn oil, and 33% lard And high - fat and high - sugar diets. The diarrhea seeds were fed diets containing 1% ethanol extract of high - fat and high - sugar diet for 12 weeks. After 12 weeks from the start of dietary intake, the animals were fasted for 12 hours and sacrificed by cardiac collection. Blood was centrifuged at 3,000 × g for 15 minutes, serum was isolated, and serum and liver tissues were stored at 70 ° C.

Dietary composition Materials Group (%) Control group High fat, high blood sugar Darya Seon-gun Casein 20.00 20.00 20.00 Corn starch 65.00 10.10 9.10 Sucrose - 27.00 27.00 alpha -cellulose 5.00 - - Corn oil 5.00 3.00 3.00 Lard - 33.00 33.00 Vitamin mixture ^{1 )} 1.00 1.40 1.40 Mineral mixture ²⁾ 3.50 5.00 5.00 D, L-methionine 0.30 0.30 0.30 Colin By Tartrate 0.20 0.20 0.20 4- butylhydroquinone ³⁾ 0.001 0.007 0.007 Purple net extract - - One

¹⁾ AIN-76 vitamin mixture

²⁾ AIN-76 mineral mixture

³⁾ Antioxidative agent, 0.01 g / 50 g lipids

② Measurement of changes in body weight and dietary intake

The body weight and the dietary intake of the control, high fat, high sugar, and Dawaya seeds are shown in Table 2 below. Body weight of the high fat and high fat diet group was significantly higher than that of the control group. Body weight of the high fat diet group and the high fat diet group were not significantly different from those of the high fat diet group. There was no significant difference in dietary intake among the three groups.

Weight and Dietary Intake Control group High-fat, high-gang Treated group Initial BW (g) 19.4 ± 2.2 19.5 ± 2.3 19.6 ± 2.3 Final BW (g) 27.0 + - 2.4 ^a 38.9 ± 3.9 ^b 37.5 ± 2.3 ^b Dietary intake
(g / day) 3.4 ± 0.4 3.0 ± 0.4 3.1 ± 0.3

< Example  3>

Measuring effect of glucose control effect and insulin resistance improvement

<3-1> Serum Glucose  And insulin concentration measurement

Blood glucose was measured by glucose oxidase enzyme method. That is, 3 mL of a coloring reagent prepared by mixing enzyme solution and buffer solution 120 mL (ASAN PHARMACEUTICAL, Korea) was added to 20 μL of plasma and reacted at 37 ° C. for 5 minutes to develop color. Then, using a spectrophotometer The absorbance was measured at 500 nm and the blood glucose concentration was calculated by substituting the following equation.

Serum insulin concentrations were measured using a mouse insulin ELISA kit (Mercodia AB, Sweden) and the method was measured according to the manufacturer's instructions.

<3-2> Measurement of insulin resistance improvement effect

The homeostasis model assessment for insulin resistance (HOMA-IR) was used as an index of insulin resistance. HOMA-IR was calculated from [fasting blood glucose (mmol / L) x fasting insulin (uU / mL) /22.5].

As a result, the blood glucose level of the high fat and high glucose groups was 165.0 ± 19.4 mg / dL, which was significantly higher than that of the control group (122.2 ± 15.7 mg / dL) (137.0 ± 17.6 mg / dL), and there was no significant difference between the control group and the high-fat and high-glycemic group (see FIG. 1). Serum insulin concentrations of control group, high fat, high sugar group, and Durae group were 18.8 ± 4.1, 37.3 ± 5.1, and 27.3 ± 3.6 μU / mL, respectively. Serum insulin concentration of high fat and high glucose group was significantly increased compared with control group. However, the serum insulin concentration in the group that ingested the extracts of Dulaya extract was significantly lower than that of the high fat and high sugar groups (see FIG. 2).

The HOMA-IR of the high-fat and high-fat group was 15.1 ± 2.7, which was significantly higher than that of the control group (5.6 ± 1.3). The HOMA-IR of the Dahlia group was 9.3 ± 2.2, which was significantly lower than that of the high fat and high sugar groups. In the C57BL / 6J mice, high-fat and high-sugar diets induced insulin resistance and increased hyperinsulinemia, HOMA-IR, and hyperglycemia. However, consumption of the ethanol extract of Dianaea reduced the insulin resistance index, HOMA-IR And improved hyperinsulinemia and hyperglycemia (see FIG. 3). Therefore, it was shown that the intake of dalasa net was helpful for the improvement of insulin resistance.

< Example  4>

Total lipid and triglyceride content of liver tissue

<4-1> Measurement of total lipid content in liver tissue

Liver tissues were obtained from mouse groups corresponding to each experimental group of Example 2 to extract lipids from liver tissues and the collected liver tissues were stored at -70 ° C. until analysis. Liver tissue was extracted by Folch method, one of crude fat analysis methods, and used for lipid measurement. 0.1 g of liver tissue was homogenized by adding chloroform and methanol (2: 1), shaking, and distilled water (15 mL) was added to the separating funnel to separate the layers. After separation, the supernatant was discarded and the mixture was diluted with chloroform and methanol (8: 4: 3) to separate the remaining chloroform layer. After separation of the layers, the concentrate was concentrated in a vacuum concentrator (Evaporator) 10 mL was added to extract lipids from liver tissue. After that, the aluminum plate was heated in an oven at 80 ° C. for 30 minutes and then cooled. The resulting chloroform layer solution (5 mL) was placed on a plate, placed on a hot plate and volatilized. The plate was heated again for 30 minutes, and the fat weight was measured by weighing.

As shown in FIG. 4, total liver lipid contents of the control, high fat, high sugar, and Darae seeds were 79.7 ± 14.9, 133.9 ± 20.2, and 108.6 ± 16.4 mg / g liver, respectively. That is, the total lipid content of the liver tissues of the high fat and high glucose groups was significantly higher than that of the control group, whereas the total lipid contents of the liver tissues of the high fat and high glucose groups were significantly lower than those of the high fat and high glucose groups , And there was no significant difference from the control group.

<4-2> Measurement of triglyceride content in liver tissue

The content of triglyceride in the extracts was determined by enzymatic method using a kit for quantification by enzymatic method (Asan Pharmaceutical, Korea). 0.1 ml of the extract of the extract was applied to a dry bath (DW-110, Daeil Tech, Korea), and the solvent was completely volatilized and dissolved in 0.2 ml of ethanol. To prevent turbidity with the reaction solution, 3 ml of the reaction reagent in which 0.5 ml of triton X-100 was added was added to the buffer solution, and the solution was developed at 37 ° C for 5 minutes and absorbance was measured at 550 nm. The absorbance value of the sample was substituted into the following equation to calculate the triglyceride content of liver tissue.

The statistical analysis of all the results of the present invention was expressed as mean ± standard error. The significance test between the groups was performed using the one-way ANOVA and the Tukey's test was used as the post test (P <0.05)

As shown in FIG. 5, in the case of high fat and high glucose (39.7 ± 5.5 mg / g liver), liver lipid accumulation was significantly increased compared with the control (16.1 ± 2.8 mg / g liver) Respectively. However, the content of triglyceride (28.9 ± 5.5 mg / g liver) in liver tissue of Dahlia group was significantly lower than that of high fat and high sugar group and there was no significant difference between the groups.

These results indicate that the long-term intake of high-density polysaccharide in high-fat and high-sugar diets induced in insulin-resistant and non-alcoholic fatty liver can significantly reduce the total lipid and triglyceride content of liver tissue Thus, ultimately, it could be shown that sucrose can be effective in the improvement and treatment of nonalcoholic fatty liver disease.

< Example  5>

Dalae  Analysis of anti-inflammatory activity of extract

Furthermore, the present inventors investigated whether the extracts of P. xylostella influenced the production of TNF-α, MCP-1 and IL-6, which are known as inflammatory cytokines, Serum concentrations of TNF-α, MCP-1 and IL-6 in the serum were measured using mouse ELISA kit (eBioscience, USA) , And the method was measured according to the manufacturer's instructions.

Serum TNF-α concentrations (32.4 ± 6.3 pg / mL) were significantly increased in the high-fat and high-fat diet groups compared to the control group (16.7 ± 3.6 pg / mL) Serum TNF-α levels were significantly decreased (23.3 ± 4.2 pg / mL) compared to the control group (see FIG. 6). The serum MCP-1 levels were 28.5 ± 4.2, 44.2 ± 7.1, and 40.7 ± 7.9 pg / mL, respectively, in the control, high fat, high glucose and high glucose groups. Serum MCP- , And the intake of Mulberry seeds significantly decreased the serum MCP-1 concentration compared to the high fat / high sugar group (see FIG. 7). Long-term intake of high-fat and high-fat diets increased serum IL-6 levels (17.1 ± 3.8 pg / mL) compared with the control group (5.7 ± 1.2 pg / mL) -6 concentration was significantly lower than that of the high fat / high sugar group (see FIG. 8).

On the other hand, as described above, TNF-α, MCP-1 and IL-6 are known as cytokines that promote inflammation. Obesity induces changes in the adipocytokine secretion of adipocytes, leading to elevated levels of inflammatory cytokines and inducing persistent inflammatory responses, leading to insulin resistance. Therefore, the intake of high-fat and high-sugar diets can induce low degree inflammation by increasing the concentration of IL-6 and MCP-1 in the serum, but the extract of the present invention has an action to reduce the production of inflammatory cytokines It can be shown that it may be effective in reducing insulin resistance and improving and treating non-alcoholic fatty liver.

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (14)

A pharmaceutical composition for the treatment of non-alcoholic fatty liver disease comprising Actinidia arguta shoot extract as an active ingredient,
The method of claim 1 or 2, wherein the extract of Phellinus Liliaceae is extracted by adding distilled water to the plant extracts, freeze-drying the distilled water extract, extracting the lyophilized product with ethanol, filtering the ethanol extract, and concentrating the filtrate under reduced pressure , &Lt; / RTI &gt;
The net extract of the genus Xeraya is contained in an amount of 0.1 to 10% by weight based on the total weight of the composition,
The net extract of the present invention has the effect of reducing the blood glucose and serum insulin in the body due to the ingestion of high fat and high sugar and reducing the lipid content and triglyceride content of liver tissue,
The extracts of the present invention have anti-inflammatory activity by inhibiting the production of inflammatory cytokines TNF-a, MCP-1 and IL-6,
Wherein the non-alcoholic fatty liver disease is selected from the group consisting of non-alcoholic simple fatty liver disease, non-alcoholic fatty liver disease, and non-alcoholic liver cirrhosis. delete delete delete delete delete delete A health functional food for improving non-alcoholic fatty liver disease comprising Actinidia arguta shoot extract as an active ingredient,
The method of claim 1 or 2, wherein the extract of Phellinus Liliaceae is extracted by adding distilled water to the plant extracts, freeze-drying the distilled water extract, extracting the lyophilized product with ethanol, filtering the ethanol extract, and concentrating the filtrate under reduced pressure , &Lt; / RTI &gt;
The net extract of the genus Xeraya is contained in an amount of 0.1 to 10% by weight based on the total weight of the composition,
The net extract of the present invention has the effect of reducing the blood glucose and serum insulin in the body due to the ingestion of high fat and high sugar and reducing the lipid content and triglyceride content of liver tissue,
The extracts of the present invention have anti-inflammatory activity by inhibiting the production of inflammatory cytokines TNF-a, MCP-1 and IL-6,
The non-alcoholic fatty liver disease is selected from the group consisting of non-alcoholic simple fatty liver, non-alcoholic fatty liver and non-alcoholic liver cirrhosis. delete delete delete delete delete delete

Images