WO2018072276A1

WO2018072276A1 - Health-care food and preparation method therefor

Info

Publication number: WO2018072276A1
Application number: PCT/CN2016/109045
Authority: WO
Inventors: 杜正彩; 邓家刚; 侯小涛; 郝二伟; 夏中尚; 黄丽贞; 谢滟
Original assignee: 广西中医药大学
Priority date: 2016-10-19
Filing date: 2016-12-08
Publication date: 2018-04-26
Also published as: CN106562415A; AU2016313671A1

Abstract

A health-care food consisting of the following raw materials in parts by weight: 5-30 parts of radix puerariae extract; 2-20 parts of gynostemma pentaphylla extract; 0.5-8 parts of radix notoginseng extract; 1-20 parts of fructus mori extract; 1-10 parts of clam meat extract; and 2-20 parts of spirulina extract. According to the health-care food and the preparation method therefor, Chinese traditional medicines of radix puerariae, gynostemma pentaphylla, fructus mori and radix notoginseng are compatible with clam meat and spirulina to prepare the health-care food which can aid in reducing blood glucose and diabetic complications; and the health-care food has remarkable blood glucose reduction effect, is safe and has no toxic and side effects.

Description

Health food and preparation method thereof

Technical field

The invention relates to the technical field of health foods, in particular to a health food with hypoglycemic function and a preparation method thereof.

Background technique

Diabetes mellitus (DM) is a non-infectious chronic endocrine metabolic disease with complications. With the development of social economy and the changes in people's diet, the prevalence of diabetes has increased year by year, seriously endangering human health. On December 1, 2015, the newly released data of the International Diabetes Federation (IDF)'s authoritative guide, Diabetes Atlas, shows that the current number of adult diabetes in the world has reached 415 million, while the number of adult diabetes patients in China has reached 110 million. Become the world's largest "diabetes" in diabetes.

Diabetes can occur at any age. With the prolongation of the disease, it is easy to be complicated with systemic nerves, microvessels, and large blood vessels. It can lead to chronic progressive lesions of the heart, brain, kidney, nerves, and eyes. Many complications are increasing. The degree of aggravation is a serious threat to the health and life of patients. At present, most of the drugs for treating diabetes are focused on stimulating insulin production in islets, and the sensitivity of insulin target cells to insulin has not been fundamentally improved, so once the drug is stopped for a while, the blood sugar often rebounds again. At the same time, due to the stimulation of islets by long-term medication, the islets are in a long-term intense working state, which eventually leads to the fatigue of islet function, so that diabetics eventually have to use insulin in vitro to help reduce blood sugar, which will further lead to the part of islet function or All of them are lost. At the same time, due to the repeated rebound of blood sugar, the viscosity of the blood is increased, and the microcirculation of the blood is impeded. Finally, complications such as heart, brain, kidney and skin nerves occur.

Summary of the invention

In view of the above, an object of the present invention is to provide a health food and a preparation method thereof to overcome the deficiencies of the prior art.

Based on the above object, the active ingredient of the health food provided by the present invention is composed of the following raw materials in parts by weight:

5 to 30 parts of pueraria;

2 to 20 strands of Gynostemma pentaphyllum;

0.5 to 8 parts of Sanqi;

1 to 20 parts of mulberry;

1 to 10 pieces of meat; and

2 to 20 parts of spirulina.

In some embodiments of the present invention, the active ingredient of the health food is composed of the following raw materials in a weight ratio:

7 to 25 parts of Pueraria;

5 to 15 parts of Gynostemma pentaphyllum;

1 to 5 copies of Sanqi;

4 to 15 parts of mulberry;

2 to 8 pieces of meat; and

4 to 15 parts of spirulina.

As still another embodiment of the present invention, the active ingredient of the health food provided by the present invention is composed of the following raw materials in a weight ratio:

5 to 30 parts of Pueraria extract;

2 to 20 parts of Gynostemma pentaphyllum extract;

0.5 to 8 parts of the extract of Panax notoginseng;

1 to 20 parts of the extract of mulberry;

1 to 10 extracts of clam meat; and

2 to 20 parts of spirulina extract.

7 to 25 parts of Pueraria extract;

5 to 15 parts of Gynostemma pentaphyllum extract;

1 to 5 parts of the extract of Panax notoginseng;

4 to 15 parts of the extract of mulberry;

2 to 8 extracts of clam meat; and

4 to 15 parts of Spirulina extract.

In some embodiments of the invention, the health food is used to prepare a hypoglycemic effect Healthy food or medicine, and safe and non-toxic side effects.

The invention also provides a preparation method of the above health food, comprising the following steps:

Mixing Pueraria and Gynostemma pentaphyllum, first adding alcohol for reflux extraction, then concentrating, drying, and then pulverizing to obtain extract I;

After mixing mulberry, clam meat and spirulina, first add water for boiling extraction, then concentrate, add ethanol to the concentrated liquid, filter and take the precipitate, dry and pulverize to obtain extract II;

Beat the Sanqi into a fine powder and use the Sanqi powder for later use;

The extract I, the extract II and the notoginseng powder were mixed to obtain the health food.

In some embodiments of the invention, the method for preparing the extract I comprises the following steps:

Mix Pueraria and Gynostemma pentaphyllum, first add alcohol for reflux extraction, add 6-10 times of alcohol each time, extract for 1 to 2 hours each time, extract 1~4 times, combine the extracts, concentrate and dry, and dry at 80 to 90 ° C, pulverized through 80 to 120 mesh sieve to obtain extract I.

Mix Pueraria and Gynostemma pentaphyllum, add 6-10 times of alcohol for the first time, soak for 30~60min, then extract by reflux, add 6~8 times of alcohol for the second time, extract for 1~2 hours each time, combine and extract The solution was concentrated, dried, dried at a temperature of 80 to 90 ° C, and pulverized through a sieve of 80 to 120 mesh to obtain an extract I.

In some embodiments of the invention, the method of preparing the extract II comprises the following steps:

Mix mulberry, clam meat and spirulina, first add water for boiling extraction, extract 1~4 times, add 6~10 times of water each time, extract for 1~2 hours each time, combine the decoction and concentrate;

Ethanol precipitation was added to the concentrate for 16 to 30 hours, and the precipitate was taken after filtration, dried and pulverized to obtain Extract II.

Mix mulberry, clam meat and spirulina, add 6~10 times of water for the first time, soak for 30~60min, then heat to boiling extraction, add 5~9 times of water for the second time, each extraction 1 to 2 hours, combined with decoction and concentrated;

To the concentrate, a volume concentration of 80 to 95% of ethanol to ethanol is added to a volume concentration of 70 to 90%, and then precipitation is carried out for 16 to 30 hours. After filtration, the precipitate is taken, dried at 60 to 75 ° C, and pulverized through 80 to 120 mesh. Sieve to obtain extract II.

In some embodiments of the invention, the decoction is concentrated to d = 1.0 to 1.5 at 80 °C.

It can be seen from the above that the health food provided by the present invention and the preparation method thereof are prepared by combining the traditional Chinese medicine Pueraria, Gynostemma pentaphyllum, Mulberry, Sanqi and Wenyu meat, and Spirulina to prepare a health food for lowering blood sugar and reducing diabetic complications. The health food has the function of assisting blood sugar lowering, and has safe and non-toxic side effects.

detailed description

In order to make the objects, the technical solutions and the advantages of the present invention more comprehensible, the present invention will be further described in detail below in conjunction with the specific embodiments.

Traditional Chinese medicine has a long history of diabetes prevention and control, and has accumulated rich experience and formed a unique theoretical understanding. Traditional Chinese medicine refers to diabetes as a form of diabetes. Many effective prescriptions can significantly improve the symptoms of “three more and one less” in diabetic patients, and can effectively reduce the complications of diabetes. After long-term experimental research, we have prepared traditional Chinese medicine Pueraria, Gynostemma, Mulberry, Sanqi and Wenyu meat, Spirulina to help reduce blood sugar and reduce diabetes complications from the perspective of traditional TCM syndrome differentiation and modern chemical composition and biological activity research. Health food, and safe and non-toxic side effects to make up for the shortcomings of the prior art.

Example 1: Preparation of health food capsules

(1) Mix 20 kg of Pueraria and 12 kg of Gynostemma pentaphyllum, add 8 times the volume concentration of 75% edible alcohol for the first time, soak for 30 minutes, then extract for 1 hour under reflux, and add volume of 6 times for the second time. 75% edible alcohol, reflux extraction for 1 hour, after extraction 2 times, the extract (combined extract) is concentrated and dried, drying temperature is 85 ° C, crushed through a 100 mesh sieve to obtain extract I;

(2) first soak 3.5 kilograms of clam meat with appropriate amount of water for 20 hours, then mix with 6 kilograms of mulberry and 15 kilograms of spirulina, add 8 times of water for the first time, soak for 30 minutes, then heat to boiling extraction. 1 hour, the second time added 6 times the amount of water, heated to boiling extraction for 2 hours, combined with 2 decoctions (filtered), the decoction was concentrated to d≈1.2 (80 ° C); volume concentration was added to the concentrate 85% ethanol to ethanol volume concentration of 75%, and then precipitated for 24 hours, filtered, the precipitate (polysaccharide), dried in an oven at 75 ° C, crushed through a 100 mesh sieve to obtain extract II;

(3) Put 2 kg of Sanqi into powder, pass through a 100 mesh sieve, and sterilize with damp heat to obtain Panax notoginseng powder;

(4) The extract I, the extract II and the notoginseng powder were mixed for 30 min, and then uniformly mixed with suitable excipients, and loaded with 0# capsule, 0.5 g/granule, 60 capsules/bottle. The health food is obtained by sealing and labeling.

Example 2: Preparation of health food granules

(1) Mix 8 kg of Pueraria and 10 kg of Gynostemma pentaphyllum, add 6 times the volume concentration of 78% edible alcohol for the first time, soak for 45 minutes, then reflux for 1.5 hours, and add the volume concentration of 8 times for the second time. For 70% edible alcohol, reflux extraction for 2 hours, after extraction 2 times, the extract (combined extract) is concentrated and dried, drying temperature is 88 ° C, crushed through a 100 mesh sieve to obtain extract I;

(2) first soak 8 kg of simmered meat with appropriate amount of water for 24 hours, then mix with 11 kg of mulberry and 9 kg of spirulina, add 7 times of water for the first time, soak for 55 minutes, then heat to boiling extraction. 1.2 hours, the second time added 9 times the amount of water, heated to boiling extraction for 1 hour, combined with 2 decoctions (filtered), the decoction was concentrated to d≈1.3 (80 ° C); volume concentration was added to the concentrate The volume concentration of 90% ethanol to ethanol is 86%, and then precipitated for 28 hours, filtered, and the precipitate (polysaccharide) is taken, dried in an oven at 65 ° C, and pulverized through a 120 mesh sieve to obtain an extract II;

(3) Beating 3.5 kg of Sanqi into powder, passing through 100 mesh sieve, and sterilizing by damp heat to obtain Panax notoginseng powder;

(4) The extract I, the extract II and the notoginseng powder are mixed, and then uniformly mixed with a suitable auxiliary material, and the granules are prepared according to the preparation method, and the health food is obtained by dispensing.

Example 3: Preparation of health food tablets

(1) Mix 5 kg of Pueraria and 6 kg of Gynostemma pentaphyllum, add 6 times the volume concentration of 70% edible alcohol for the first time, soak for 40 minutes, then reflux for 1.2 hours, and add the volume concentration of 8 times for the second time. It is 75% edible alcohol, reflux extraction for 2 hours, the third addition of 7 times the amount of 75% edible alcohol, reflux extraction for 1 hour, extraction 3 times, the extract (combined extract) is concentrated and dried, drying temperature is 88 ° C , crushed through a 80 mesh sieve to obtain extract I;

(2) first soak 1.5 kg of clam meat with appropriate amount of water for 18 hours, then mix with 15 kg of mulberry and 11 kg of spirulina, add 6 times of water for the first time, soak for 30 minutes, then heat to boiling extraction. 1.3 hours, the second addition of 5 times the amount of water, heated to boiling extraction for 1 hour, combined with 2 decoctions (filtered), the decoction was concentrated to d≈ 1.15 (80 ° C); volume concentration was added to the concentrate 92% ethanol to ethanol volume concentration of 84%, and then precipitated for 20h, filtered, the precipitate (polysaccharide), dried in an oven at 65 ° C, crushed through a 110 mesh sieve to obtain extract II;

(3) Put 2.8 kg of Sanqi into powder, pass through a 120 mesh sieve, and sterilize with damp heat to obtain Panax notoginseng powder;

(4) The extract I, the extract II and the notoginseng powder are mixed for 40 minutes, and then uniformly mixed with a suitable auxiliary material, compressed, and packaged to obtain the health food.

Example 4: Preparation of health food ointment

(1) Mix 26 kg of Pueraria and 9 kg of Gynostemma pentaphyllum, add 10 times the volume concentration of 70% edible alcohol for the first time, soak for 60 minutes, then extract for 1 hour under reflux, and add volume of 7 times for the second time. For 70% edible alcohol, reflux extraction for 1 hour, after extraction 2 times, the extract (combined extract) is concentrated and dried, drying temperature is 80 ° C, crushed through 80 mesh sieve, to obtain extract I;

(2) Soak 7.5 kg of clam meat with appropriate amount of water for 22 hours, then mix with 13 kg of mulberry and 18 kg of spirulina, add 9 times of water for the first time, soak for 38 minutes, then heat to boiling extraction. 1.6 hours, the second time adding 7 times the amount of water, heating to boiling extraction for 1.5 hours, the second time adding 8 times the amount of water, heating to boiling extraction for 1 hour, combining 3 times of decoction (filtration), the decoction Concentrated to d≈1.25 (80 ° C); a volume concentration of 80% ethanol to ethanol was added to the concentrate at a volume concentration of 72%, and then precipitated for 29 hours, filtered, and the precipitate (polysaccharide) was taken and dried in an oven at 68 ° C. , crushed through a 100 mesh sieve to obtain extract II;

(3) Put 4.5 kg of Sanqi into powder, pass through a 120 mesh sieve, and sterilize with damp heat to obtain Panax notoginseng powder;

(4) The extract I, the extract II and the notoginseng powder are mixed, and then uniformly mixed with a suitable auxiliary material, and the paste is prepared according to a preparation method, and the package is obtained to obtain the health food.

Example 5: Preparation of health food oral liquid

(1) Mix 14 kg of Pueraria and 13 kg of Gynostemma pentaphyllum, add 7.5 times the volume concentration of 80% edible alcohol for the first time, soak for 44 minutes, then reflux for 1.8 hours, and add a volume of 6 times for the second time. 75% edible alcohol, reflux extraction for 2 hours, after extraction 2 times, the extract (combined extract) is concentrated and dried, drying temperature is 90 ° C, crushed through a 120 mesh sieve to obtain extract I;

(2) first soak 7 kg of clam meat with appropriate amount of water for 17 hours, then mix with 10 kg of mulberry and 5 kg of spirulina, add 7 times of water for the first time, soak for 55 minutes, then heat to boiling extraction. 1.7 hours, the second time added 5.5 times the amount of water, heated to boiling extraction for 2 hours, combined with 2 decoctions (filtered), the decoction was concentrated to d≈1.2 (80 ° C); volume concentration was added to the concentrate The volume concentration of 93% ethanol to ethanol is 87%, and then precipitated for 20 hours, filtered, and the precipitate (polysaccharide) is taken, dried in an oven at 72 ° C, and pulverized through a 100 mesh sieve to obtain an extract II;

(4) The extract I, the extract II and the notoginseng powder are mixed for 30 minutes, and then uniformly mixed with a suitable auxiliary material, and the oral liquid is prepared according to the preparation method, and the health food is obtained by encapsulation.

Example 6: Preparation of health food pellets

(1) Mix 15 kg of Pueraria and 13 kg of Gynostemma pentaphyllum, add 6.5 times the volume of the first time to 76% edible alcohol, soak for 45 minutes, then extract for 1.9 hours under reflux, and add 7.5 times the volume concentration for the second time. For 72% edible alcohol, reflux extraction for 1.1 hours, after extracting 2 times, the extract (combined extract) is concentrated and dried, drying temperature is 82 ° C, crushed through a 100 mesh sieve to obtain extract I;

(2) first soak 9 kg of clam meat with appropriate amount of water for 24 hours, then mix with 3 kg of mulberry and 9 kg of spirulina, add 6 times of water for the first time, soak for 58 minutes, then heat to boiling extraction. 1 hour, the second time adding 5.5 times the amount of water, heating to boiling extraction for 1 hour, the third time adding 5.5 times the amount of water, heating to boiling extraction for 1.5 hours, the fourth time adding 5 times the amount of water, heating to boiling After extracting for 1.2 hours and combining 4 times of decoction (filtration), the decoction was concentrated to d≈1.1 (80 ° C); a volume concentration of 78% ethanol to ethanol was added to the concentrate to a volume concentration of 70%, and then precipitated. After 18 hours, the mixture was filtered, and the precipitate (polysaccharide) was taken, dried in an oven at 73 ° C, and pulverized through a 100 mesh sieve to obtain an extract II;

(3) Dilute 4 kg of Sanqi into a powder, pass through a 100 mesh sieve, and sterilize with damp heat to obtain Panax notoginseng powder;

(4) The extract I, the extract II and the notoginseng powder are mixed, and then uniformly mixed with a suitable auxiliary material, and the pellet is prepared according to the preparation method, and the package is obtained to obtain the health food.

Auxiliary hypoglycemic function animal experiment

1 Materials and methods

1.1 Sample: Gesang hypoglycemic capsule, specification: 0.5g / grain, product batch number: 150201, stored in a dry place. The recommended dosage for oral administration is 2 times per person (adult) per person, 3 capsules per time, and the adult weight is calculated as 60kg, and the equivalent dose is 50mg/kg BW. Capsule contents were taken for experiments.

1.2 Experimental animals: SPF-class healthy adult Kunming male mice 144 were selected from Guangdong Medical Experimental Animal Center.

Only, the weight is 24 ~ 28 grams, the experimental animal production license number: SCXK (Gui) 2013-0002, laboratory animal quality certificate number: 44007200027357.

1.3 Experimental animal room environmental conditions: animal laboratory is a barrier system, use license number: SYXK (Gui) 2011-0005.

Animal laboratory temperature: 22 ~ 25 ° C, relative humidity: 55 ~ 70%.

1.4 Dose selection and administration of the test substance: According to the recommended dosage of the sample, 250, 500, 1000 mg/kg BW

Three doses of the test group (5, 10, 20 times the recommended amount of the human body), and a model control group, a test sample group (high dose) and a blank control group for each normal animal. Group 12 mice. Weigh 1.25, 2.50, 5.00g of Gesang Jiangtang Capsules, add pure water to 100mL, mix well, and mix them into 12.5, 25.0, 50.0mg/mL concentration suspension, respectively, and give the corresponding dose groups to the stomach, normal The test sample group of animals was given a concentration of 50.0 mg/mL suspension, and the volume of gastric perfusion was 0.2 mL/10 gBW. The model control group and the normal animal blank control group were given an equal volume of pure water, once a day, and continuously administered by stomach. 30 days.

1.5 main instruments and reagents

Instrument: Luo Kangquan excellent blood glucose meter, electronic analysis of the same day.

Reagents: blood glucose test strips, etc.

1.6 Experimental method: According to the National Food and Drug Administration issued the National Food and Drug Administration [2012] No. 107 "Notice on Printing and Evaluation of Antioxidant Function Evaluation Methods, 9 Applicable Methods for Evaluation of Health Function", Annex 3 - Auxiliary Hypoglycemic Function The evaluation method uses an animal test protocol--islet damage high blood glucose model method.

1.6.1 Hyperglycemia model animal modeling

The mice were given an intraperitoneal injection of alloxan (120 mg/kg BW) 24 hours after fasting. After 5 days, fasting for 4 hours, blood is taken from the eye to measure blood sugar. If the blood glucose level is 10 to 25 mmol/L, the model for making high blood sugar can be considered successful. 120 mice were selected for modeling, and 102 of them were successfully modeled. 96 mice with successful modeling were included in the experiment, and the mice were divided into 2 batches. The first batch of 48 patients were tested for fasting blood glucose lowering, and the second batch of 48 mice were tested for glucose tolerance.

1.6.2 Reduce the fasting blood glucose test

The first batch of hyperglycemic model animals were randomly divided into 4 groups according to the fasting 4 hours blood glucose level, 12 in each group. The test group was given different concentrations of sample solution, and the model control group was given pure water for 30 consecutive days. The fasting blood glucose level was measured by blood sampling (fasting before the test), and the fasting blood glucose level and percentage of blood glucose decreased in each group were compared.

Percentage of blood glucose drop = (pre-experiment blood glucose value - post-experiment blood glucose value) / pre-experiment blood glucose value x 100%.

Under the condition of model establishment, if the measured value of fasting blood glucose in the experimental group is lower than the model control group or the percentage of blood glucose drop is statistically significant, the test result can be judged as positive.

1.6.3 Normal animals: 24 healthy adult animals were selected and grouped according to fasting blood glucose levels for 4 hours, randomly divided into blank control groups.

And the test sample group (high dose), 12 in each group. The rest of the operation is the same as 1.6.2.

1.6.4 Glucose tolerance test: 48 rats of the second batch of hyperglycemic model mice were randomly divided into 4 groups according to the fasting 4 hours blood glucose level, 12 in each group, and the gavage method was the same as 1.6.2. Mice were fasted for 4 hours on day 31, the test group was given different concentrations of sample solution, and the model control group was given an equal volume of pure water. After 15 minutes, each group was orally administered with a solution of glucose (2.0 g/kg BW). The blood glucose levels at 0, 0.5, and 2 hours after glucose administration were measured, and the changes in the area under the blood glucose curve at each time point after the glucose concentration of the model control group and the test sample group were observed.

Area under the blood glucose curve = 0.25 × (0 hour blood glucose level + 4 × 0.5 hour blood glucose value + 3 × 2 hours blood glucose value)

Under the condition of the model establishment, if the area under the blood glucose curve of the test group at 0, 0.5, and 2 hours after glucose administration is lower than the model control group and has statistical significance, the test result can be judged as positive.

1.7 Test data processing: Statistical analysis of variance analysis was performed using SPSS11.0 statistical software.

1.8 Result judgment: Under the condition of model establishment, one of the two indicators of fasting blood glucose and glucose tolerance is positive, and it has no effect on the fasting blood glucose of normal animals. It can be determined that the test sample is positive for the blood glucose-reducing animal.

2 test results

2.1 The effect of sample on mouse body weight

The initial body weight, mid-term body weight, terminal body weight and weight gain of the mice in each dose group of hyperglycemic model were not significantly different from those in the model control group (P>0.05). See Table 1; mice in the normal animal experimental group. The initial body weight, mid-term weight, terminal body weight and weight gain were not significantly different from the blank control group (P>0.05), as shown in Table 2. The results showed that the sample had no significant effect on the weight gain of hyperglycemic model mice and normal mice.

Table 1. Effect of Gesang Jiangtang Capsule on body weight of hyperglycemic model mice

Note: The initial body weight, mid-term weight, terminal body weight and weight gain of the mice in each dose group were not significantly different from the model control group (P>0.05).

(Continued) Table 1. Effect of Gesang Jiangtang Capsule on body weight of hyperglycemic model mice

Table 2. Effect of Gesang Jiangtang Capsule on normal mouse weight

Note: The initial body weight, mid-term body weight, terminal body weight and weight gain of the experimental group were not significantly different from the blank control group (P>0.05).

2.2 Effect of sample on fasting blood glucose in hyperglycemic model mice

It can be seen from Table 3 that the blood glucose level after fasting is >10 mmol/L after 4 hours of modeling, and the model is established. The mice were given different doses of Gesang Jiangtang Capsules for 30 days. The blood glucose levels of the 30th day of each dose group were lower than the model control group. The blood glucose drop value and percentage decrease of each dose group were higher than the model control group, and high. The blood glucose level, blood glucose decrease value and percentage decrease of the middle dose group were significantly different from the model control group (P<0.01 or P<0.05), indicating that the sample has the effect of lowering the fasting blood glucose of the hyperglycemic model mice.

Table 3. Fasting blood glucose results in hyperglycemic model mice

Note: * indicates that compared with the model control group, P < 0.05, ** indicates comparison with the model control group, P < 0.01.

2.3 Effect of sample on glucose tolerance test in hyperglycemic model mice

Table 4. Results of glucose tolerance test in hyperglycemic model mice

Note: ** indicates P<0.01 compared with the model control group.

It can be seen from Table 4 that after 30 days of oral administration of different doses of Gesang Jiangtang Capsules in hyperglycemic model mice, the area under the blood glucose curve of each dose group was lower than that of the model control group, and the high and middle dose groups and the model control group. The difference was very significant (P<0.01), indicating that the sample has the effect of reducing the area under the blood glucose curve at each time point in the glucose tolerance test of hyperglycemic mice, and the glucose tolerance test results are positive. .

2.4 Effect of sample on fasting blood glucose in normal mice

As can be seen from Table 5, there was no significant difference in the fasting blood glucose level between the mice and the normal control group (P>0.05). Fasting blood glucose in normal mice had no significant effect.

Table 5. Fasting blood glucose results in normal mice

Note: There was no significant difference in blood glucose values between the sample group before and after the test and the blank control group (P>0.05).

3 Conclusion:

Gesang Jiangtang Capsules with 250, 500, 1000mg/kg BW (equivalent to 5, 10, 20 times the recommended amount of human body) were administered to hyperglycemic mice for 30 days, which could reduce the fasting blood glucose level of mice. To reduce the area under the blood glucose curve at the time of the glucose tolerance test in mice, the glucose tolerance test results were positive, and had no significant effect on the weight gain of the mice; the normal mice were continuously gavage for 30 days at a dose of 1000 mg/kg BW. There was no significant effect on fasting blood glucose and weight gain in mice. In summary, the sample has an auxiliary blood glucose lowering function.

Acute and chronic toxicity studies

1 Materials and methods

1.1 sample: Gesang hypoglycemic capsule, specification: 0.5g / grain, product batch number: 150201, set in the shade Store in a dry place. The recommended dosage for oral administration is 2 times per person (adult) per person, 3 capsules per time, and the adult weight is calculated as 60kg, and the equivalent dose is 50mg/kg BW.

1.2 Experimental animals and environment: SPF-grade healthy Kunming mice and SD rats were all propagated by the Experimental Animal Center of Guangxi Medical University. The experimental animal production license number: SCXK (Gui) 2014-0002, the experimental animal quality certificate number : 45000300000170, 45000300000223; 45000300000379. The animal laboratory is a barrier system, using the license number: SYXK (Gui) 2011-0005. Animal laboratory temperature: 22 ~ 25 ° C, relative humidity: 55 ~ 70%.

1.3 Acute oral toxicity test of mice: 20 mice of Kunming mice weighing 18-22 g were selected by the maximum tolerated dose (MTD) test method, half male and half female. Animals were fasted for 16 hours before the test, and were not allowed to drink water. Weigh 20.0g sample, add pure water to 40mL, mix well, prepare 500mg/mL concentration suspension, and then give the animal 2 times (interval 6h), each dose is 0.4mL/20gBW, total dose It is 20,000 mg/kg BW. After the gavage, the animal's poisoning performance was observed and recorded. Weigh once a week and observe for two weeks. At the end of the experiment, dissect the animals for general observation. The acute toxicity of the test substance was evaluated according to the toxicity classification standard.

1.4 Rat 30-day feeding test

1.4.1 Dose selection and test substance administration method: 80 SD rats were selected, female and male, and the body weight was 60-80 g. The animals were randomly divided into 4 groups, namely a negative control group and 3 test groups, 20 in each group, half male and half male. The doses of the three test groups were set to 5000, 2500, and 1250 mg/kg BW, respectively, which corresponded to 100, 50, and 25 times the recommended dose of the human body. Weigh 50.0, 25.0, 12.5g samples, add pure water to 100mL, mix well, and mix 500, 250, 125mg/mL concentration suspension, and weigh the corresponding dose group according to the volume of 1.0mL/100g BW. The negative control group was given an equal volume of pure water, and was intragastrically administered once a day for 30 days.

1.4.2 Experimental method: All animals were given normal feed during the experiment, fed in a single cage, and fed freely. Animals were observed daily for activity and growth, fed twice a week, recorded for food intake and leftovers, weekly weight, and weekly food intake and food utilization. At the end of the experiment, the animals were fasted overnight (fasting for 16 hours, not limited to drinking water), then the animals were weighed, the rats were sacrificed, 2 blood samples were taken, and a blood anticoagulation blood cell counter was used to detect Hb, RBC, WBC and its classification. PLT and the like; another blood is not anticoagulated to separate serum, and the serum AST, ALT, BUN, Cr, TC, TG, Glu, TP, Alb and other items are detected by a kit and an automatic biochemical analyzer. After blood collection, the animals were dissected and grossly observed. The liver, kidney, spleen and testicles were weighed to calculate the ratio of the body to the body. The liver, kidney, spleen, stomach, duodenum, testis and ovary were taken. Perform a histopathological examination. In the right Animals in each dose group were examined for gross lesions and biochemical indicators. Only the histopathological examination of the main organs of the high-dose group and the control group was performed. If the lesions were found, the corresponding organs and tissues of the middle- and low-dose groups were found. checking.

1.4.3 Experimental data statistics: One-way ANOVA was performed using SPSS statistical software. In the statistical analysis, the data were first tested for homogeneity of variance. If the variances were homogeneous, one-way analysis of variance was used for the overall comparison. The difference was found by Dunnett test to compare the two groups between the multiple dose groups and the control group. If the variance is not uniform, the data is subjected to appropriate variable conversion, and after the homogeneity test of the variance is satisfied, the converted data is used for statistics; if the converted data still does not meet the variance requirement, the rank sum test is used for statistical analysis.

2 results

2.1 Acute oral toxicity test

Table 6 Results of acute toxicity test of Gesang Jiangtang Capsule on mice

As can be seen from Table 6, after the mice were administered with a dose of 20,000 mg/kg BW, the animals grew well and no body weight was affected. No symptoms of poisoning were observed in the mice tested, and no animal death was observed for 14 days. At the end of the experiment, the animals were dissected and observed in general, and no obvious abnormal changes were observed in the main organs such as liver, kidney, spleen, heart, lung, stomach and intestine. The results showed that the acute oral toxicity MTD of the sample to mice was greater than 20000 mg/kg BW, and the acute oral toxicity was non-toxic.

2.2 Rat 30-day feeding test

2.2.1 General performance of animals: During the experiment, the animals in each group had good growth and development. No abnormal behavior and poisoning performance were observed in the animals, and no deaths were observed in each group.

2.2.2 Effect of sample on body weight and food utilization rate in rats

The results are shown in Table 7 to Table 8. The rats were intragastrically administered with Gesang Jiangtang Capsules at a dose of 5000, 2500, and 1250 mg/kg BW for 30 days. During the experiment period, the male and female rats in each dose group received weekly weight, weight, and weight. There was no significant difference in weekly food intake and total food intake, weekly food utilization rate and total food utilization rate compared with the control group (P>0.05), indicating that the sample had no significant effect on body weight gain and food utilization rate. .

Table 7 Effect of Gesang Jiangtang Capsule on Rat Body Weight

Note: There was no significant difference between the dose groups in the table and the negative control group (P>0.05).

Table 8 Effect of Gesang Jiangtang Capsule on total food utilization rate in rats

2.5.3 Effect of sample on blood routine indicators in rats

Table 9 Results of blood routine indicators of rats after the 30-day feeding test of Gesang Jiangtang Capsule

Table 10 Results of blood routine indicators of rats after the 30-day feeding test of Gesang Jiangtang Capsule

It can be seen from Tables 9 and 10 that Gesang Jiangtang Capsules at the doses of 5000, 2500, and 1250 mg/kg BW were administered to rats for 30 days. The hemoglobin, total number of red blood cells, total number of white blood cells and their total number of male and female rats in each dose group were There was no significant difference in the classification and platelet counts compared with the control group (P>0.05), indicating that the sample had no significant effect on the blood routine parameters of the rats.

2.5.4 Effect of sample on blood biochemical parameters in rats

The results are shown in Table 11 and Table 12. The rats were intragastrically administered with Gesang Jiangtang Capsule at 5000, 2500, 1250 mg/kg BW for 30 days. Serum aspartate aminotransferase, alanine aminotransferase and urea were used in the female and male rats in each dose group. Nitrogen, creatinine, cholesterol, triglyceride, total protein, albumin, and blood glucose were not significantly different from the control group (P>0.05), indicating that the sample had no significant effect on blood biochemical parameters in rats.

Table 11 Results of blood biochemical indicators in rats after the 30-day feeding test of Gesang Jiangtang Capsule

Table 12 Results of blood biochemical indicators in rats after the 30-day feeding test of Gesang Jiangtang Capsule

2.5.5 Effect of sample on organ weight and organ/body weight ratio in rats

Results: Gesang Jiangtang Capsules at 5000, 2500, 1250mg/kg BW were administered to rats for 30 days. The testis weight and liver/body, kidney/body of liver, kidney, spleen and male rats in each dose group The spleen/body and male testicular/body ratios were not significantly different from the control group (P>0.05), indicating that the sample had no significant effect on the organ weight and organ/body weight ratio.

2.5.6 Anatomical gross observation and histological examination results

At the end of the experiment, the animals were dissected, and no obvious lesions were observed in the animals in each group. Therefore, only the high-dose group of the sample and the main organs of the negative control group were examined for histopathological examination. The results are shown in Table 18 to Table 24. The results showed that 1 male in the high-dose group and 1 male and 1 female in the control group showed hepatocyte vacuolar degeneration in the hepatic lobules; 1 male and 1 female in the high-dose group and 1 in the control group. Hepatocyte dot necrosis was observed in the hepatic lobules of male and 2 female rats; a small amount of inflammatory cell infiltration was observed in the liver portal area of one male and two female rats in the high-dose group and the control group. In the high-dose group, there were 2 males and 1 female, and 1 male and 1 female rats in the control group showed a small amount of inflammatory cell infiltration in the renal cortex. The above tissue lesions belong to the spontaneous mild lesions of the animals, and the degree of tissue lesions of the two groups of animals is similar, so it can be ruled out by the sample, and no histopathological changes are observed in other organs, indicating that the sample is the above organ tissues of the rats. No damage.

3 Conclusion:

After administration of the sample of the present invention at a dose of 20,000 mg/kg BW to the mice, no symptoms of poisoning and death were observed in the animals. The acute oral toxicity of the samples of the present invention to the mice was greater than 20000 mg/kg BW, and the acute oral toxicity was none. Toxic level. 30-day feeding test (chronic toxicity test): Rats were continuously administered with a dose of 5000, 2500, 1250 mg/kg BW (100, 50, 25 times recommended by the human body, respectively) for 30 days. Animals during the experiment The growth and development were good. There was no significant difference in body weight, weight gain, food intake, food utilization rate, blood routine index, blood biochemical index, organ weight and organ/body weight ratio of the animals in each dose group. >0.05); gross anatomical observation and histopathological examination showed no abnormal changes associated with the sample. The sample was not seen to have toxic side effects on various observation indexes of the rats within the range of the amount of the reagent.

Three genotoxicity tests:

1.Ames test

1.1 Method: The strains of Salmonella typhimurium histidine-deficient strains TA97a, TA98, TA100 and TA102 which were identified to meet the requirements were tested. Weigh 5.0g of the sample of the invention, add pure water to 100mL, mix well, prepare a 50mg/mL concentration solution, and then dilute it 5 times (take 10mL plus pure water to 50mL), respectively, and make 10, 2, 0.4, 0.08 The solution of mg/mL concentration is treated by autoclaving (0.103Mpa 20min) for testing. Polychlorinated biphenyl-induced rat liver microsomal enzyme (S-9) was used as an in vitro metabolic activation system. Using the plate incorporation method, 0.1 mL of the test strain enrichment solution, 0.1 mL of the test solution and 0.5 mL of the S-9 mixture (when metabolic activation is required) are sequentially added to the maintained top culture medium, and then mixed and poured. On the bottom medium plate. The five test doses were 5000, 1000, 200, 40, and 8 μg/dish, respectively, with spontaneous reversion control, solvent control, and positive mutagen control. The spontaneous reversal control was the same as the sample group except that no sample was added. The solvent control replaced the sample with sterile pure water, and the rest of the conditions were the same as the sample group. Each strain of each dose group was made into 3 parallel dishes. The cells were cultured at 37 ° C for 48 hours, and the number of colonies per dish was counted. The entire set of tests was repeated twice under the same conditions. If the number of revertant colonies of the test substance increases more than twice the number of spontaneously reverted colonies and has a dose-response relationship, the mutagenesis test is positive.

1.2 Results: For the four test strains of TA97a, TA98, TA100 and TA102, the number of revertant colonies in each dose group did not exceed twice the number of spontaneously reverted colonies, and there was no dose-response relationship. It indicates that the mutagenesis test result of the present invention is negative.

2. Micronucleus test of mouse bone marrow cells

2.1 Method: The test was carried out by oral gavage twice at intervals of 24 hours. Choose a weight of 25 ~ 30g 50 Kunming mice were randomly divided into 5 groups, 10 in each group, half male and half female. The three doses in the experimental group were 10000, 5000, 2500 mg/kg BW, respectively, with pure water as the negative control and 40 mg/kg BW dose of cyclophosphamide (cp) as the positive control. Weigh 20.0, 10.0, 5.0g samples, add pure water to 40mL, mix well, prepare 500, 250, 125mg/mL concentration suspension, then weigh the animals according to the volume of 0.4mL/20g BW, negative The control group was given an equal volume of pure water, and the positive control group was given an equal volume of 2 mg/mL cyclophosphamide solution. The animals were sacrificed by cervical dislocation 6 hours after the second administration. The scaphoid bone marrow was diluted with calf serum, fixed with methanol, and stained with Giemsa. Under the light microscope, 1000 polychromatic red blood cells (PCE) were counted per animal, and the number of PCEs containing micronuclei was observed. The micronucleus rate was calculated as the micronucleus-containing PCE per thousand; and 200 polychromatic red blood cells were counted at the same time. The ratio of polychromatic red blood cells to mature red blood cells (PCE/NCE) was calculated. The statistical processing was performed by using the Poisson distribution mean comparison method in the SPSS statistical software. For example, the micronucleus rate of the experimental group was higher than that of the negative control group, and there was a significant dose-response relationship and statistical significance, which was a positive result.

2.2 Results: The micronucleus rate of bone marrow cells of mice in each dose group of the present invention was not significantly different from that of the negative control group (P>0.05), and the PCE/NCE values of each dose group were not less than that of the negative control group. %, there was no significant difference compared with the negative control group, and the micronucleus rate of the cyclophosphamide positive control group was significantly different from that of the negative control group (P<0.01), suggesting that the bone marrow of the sample of the present invention was not observed. The cells have damage and inhibition.

3. Mouse sperm abnormality test

3.1 Methods: 50 Kunming male mice weighing 25-35 g were randomly divided into 5 groups, 10 in each group. The three doses in the experimental group were 10000, 5000, 2500 mg/kg BW, respectively, with pure water as the negative control and 40 mg/kg BW dose of cyclophosphamide (cp) as the positive control. Weigh 20.0, 10.0, 5.0g samples, add pure water to 40mL, mix well, prepare 500, 250, 125mg/mL concentration suspension, then weigh the animals according to the volume of 0.4mL/20g BW, negative The control group was given an equal volume of pure water, and the positive control group was given an equal volume of 2 mg/mL cyclophosphamide solution. Gavage once a day for 5 consecutive days. The animals were sacrificed on the 30th day after the last dose, and the sperm smears of the epididymis were taken, fixed by methanol, and stained with eosin. Under an optical microscope, 1000 sperm counts were counted per animal, and the rate of sperm deformity was calculated. Wilcoxon rank sum test statistical processing in SPSS statistical software was applied. For example, the sperm deformity rate of the experimental group was higher than that of the negative control group, and it was statistically significant, and there was a dose-response relationship, which was a positive result.

3.2 Results: The sample of the present invention did not significantly change the incidence of sperm abnormality in mice. The sperm deformity rate of each sample group was not significantly different from that of the negative control group (P>0.05), while cyclophosphamide was not significantly different (P>0.05). The difference between the positive control group and the negative control group was very significant (P<0.01), suggesting that the sperm of the present invention did not cause spermatogenesis in male mice.

4 Conclusion:

Three genotoxicity tests (Ames test, mouse bone marrow micronucleus test, mouse sperm abnormality test) were negative. This indicates that the present invention is not genotoxic.

The above experimental research confirmed that the health food provided by the invention has obvious hypoglycemic health-care effect, and is safe and non-toxic.

It should be understood by those of ordinary skill in the art that the discussion of any of the above embodiments is merely exemplary, and is not intended to suggest that the scope of the disclosure (including the claims) is limited to these examples; Combinations of the technical features in the different embodiments are also possible, and there are many other variations of the various aspects of the invention as described above, which are not provided in the details for the sake of brevity. Therefore, any omissions, modifications, equivalents, improvements, etc., which are within the spirit and scope of the invention, are intended to be included within the scope of the invention.

Claims

A health food product, characterized in that the active ingredient of the health food product is composed of the following traditional Chinese medicine raw materials in parts by weight:

5 to 30 parts of pueraria;

2 to 20 strands of Gynostemma pentaphyllum;

0.5 to 8 parts of Sanqi;

1 to 20 parts of mulberry;

1 to 10 pieces of meat; and

2 to 20 parts of spirulina.
The health food according to claim 1, wherein the active ingredient of the health food is composed of the following traditional Chinese medicine raw materials in a weight ratio:

7 to 25 parts of Pueraria;

5 to 15 parts of Gynostemma pentaphyllum;

1 to 5 copies of Sanqi;

4 to 15 parts of mulberry;

2 to 8 pieces of meat; and

4 to 15 parts of spirulina.
A health food product, characterized in that the active ingredient of the health food product is composed of the following traditional Chinese medicine raw materials in parts by weight:

5 to 30 parts of Pueraria extract;

2 to 20 parts of Gynostemma pentaphyllum extract;

0.5 to 8 parts of the extract of Panax notoginseng;

1 to 20 parts of the extract of mulberry;

1 to 10 extracts of clam meat; and

2 to 20 parts of spirulina extract.
The health food according to claim 3, wherein the active ingredient of the health food is composed of the following traditional Chinese medicine raw materials in a weight ratio:

7 to 25 parts of Pueraria extract;

5 to 15 parts of Gynostemma pentaphyllum extract;

1 to 5 parts of the extract of Panax notoginseng;

4 to 15 parts of the extract of mulberry;

2 to 8 extracts of clam meat; and

4 to 15 parts of Spirulina extract.
A health food according to any one of claims 1 to 4, which can be used for preparing a health food or a medicine having hypoglycemic action, and can be prepared into any dosage form such as oral solid, semi-solid or oral liquid. And safe and non-toxic side effects.
A method for preparing a health food according to any one of claims 1 to 4, comprising the steps of:

Mixing Pueraria and Gynostemma pentaphyllum, first adding alcohol for reflux extraction, then concentrating, drying, and then pulverizing to obtain extract I;

After mixing mulberry, clam meat and spirulina, first add water for boiling extraction, then concentrate, add ethanol to the concentrate, filter and take the precipitate, dry and pulverize to obtain extract II;

Beat the three seven into powder to get the notoginseng powder;

The extract I, the extract II and the notoginseng powder were mixed to obtain the health food.
The method for preparing a health food according to claim 1, wherein the method for preparing the extract I comprises the following steps:

Mix Pueraria and Gynostemma pentaphyllum, first add alcohol for reflux extraction, add 6-10 times of alcohol each time, extract for 1 to 2 hours each time, extract 1~4 times, combine the extracts, concentrate and dry, and dry at 80 to 90 ° C, pulverized through 80 to 120 mesh sieve to obtain extract I.
The method for preparing a health food according to claim 7, wherein the method for preparing the extract I comprises the following steps:

Mix Pueraria and Gynostemma pentaphyllum, add 6-10 times of alcohol for the first time, soak for 30~60min, then extract by reflux, add 6~8 times of alcohol for the second time, extract for 1~2 hours each time, combine and extract The solution was concentrated, dried, dried at a temperature of 80 to 90 ° C, and pulverized through a sieve of 80 to 120 mesh to obtain an extract I.
The method for preparing a health food according to claim 6, wherein the method for preparing the extract II comprises the following steps:

Mix mulberry, clam meat and spirulina, first add water for boiling extraction, extract 1~4 times, add 6~10 times of water each time, extract for 1~2 hours each time, combine the decoction and concentrate;

Ethanol precipitation was added to the concentrate for 16 to 30 hours, and the precipitate was taken after filtration, dried and pulverized to obtain Extract II.
The method for preparing a health food according to claim 9, wherein the method for preparing the extract II comprises the following steps:

Soak the meat with water for 12 to 24 hours, then mix with mulberry and spirulina. Add 6 to 10 times of water for the first time. Soak for 30 to 60 minutes, then heat to boiling extraction. Add 5 for the second time. ~ 9 times the amount of water, each extraction for 1 to 2 hours, combined with decoction and concentrated;

To the concentrate, a volume concentration of 80 to 95% of ethanol to ethanol is added to a volume concentration of 70 to 90%, and then precipitation is carried out for 16 to 30 hours. After filtration, the precipitate is taken, dried at 60 to 75 ° C, and pulverized through 80 to 120 mesh. Sieve to obtain extract II.