CN108514093B

CN108514093B - Multifunctional food composition for assisting in reducing blood sugar and preparation method thereof

Info

Publication number: CN108514093B
Application number: CN201810448001.0A
Authority: CN
Inventors: 魏敏杰; 宫世强; 孟贺
Original assignee: China Medical University
Current assignee: China Medical University
Priority date: 2018-05-11
Filing date: 2018-05-11
Publication date: 2021-08-31
Anticipated expiration: 2038-05-11
Also published as: CN108514093A

Abstract

The invention belongs to the field of health-care food, and particularly relates to a multifunctional food composition with the functions of assisting in reducing blood sugar, increasing insulin sensitivity, protecting liver, improving intestinal flora and the like, and a preparation method and application thereof. The food composition comprises the following components in parts by weight: 20-40 parts of Chinese yam, 10-30 parts of radix polygonati officinalis, 2-20 parts of medlar, 2-20 parts of white poria, 1-5 parts of bitter gourd, 1-5 parts of rhizoma polygonati, 1-5 parts of hawthorn, 1-5 parts of black sesame, 1-4 parts of mulberry leaf, 1-4 parts of dandelion, 1-2 parts of radix puerariae and 1-2 parts of platycodon grandiflorum. Meanwhile, the health-care food has the functions of assisting in reducing blood sugar, increasing insulin sensitivity, protecting liver and improving intestinal flora. Provides reliable scientific basis for clinical use.

Description

Multifunctional food composition for assisting in reducing blood sugar and preparation method thereof

Technical Field

The invention belongs to the field of health-care food, and particularly relates to a multifunctional food composition with the functions of assisting in reducing blood sugar, increasing insulin sensitivity, protecting liver, improving intestinal flora and the like, and a preparation method and application thereof.

Background

Diabetes is a chronic metabolic disorder characterized by hyperglycemia due to a variety of etiologies. The etiology involves various aspects such as heredity, environment, immune inflammation and the like, and various causes lead to insulin secretion deficiency or impaired insulin resistance and glucose tolerance. Modern medicine divides diabetes into two main types, one is insulin-dependent type 1 diabetes and the other is non-insulin-dependent type 2 diabetes. Diabetes not only causes metabolic disorder of three nutrient substances, namely sugar, fat and protein, but also causes a series of secondary diseases, such as hypertension, diabetic foot, diabetic nephropathy, myocardial infarction and the like, and seriously threatens the health of human beings. The incidence of diabetes also shows a trend of increasing year by year in the global scope, and is now a healthy killer after cardiovascular and cerebrovascular diseases and tumors, especially for the middle-aged and elderly people, and more particularly for the high-incidence people of diabetes.

Diabetes necessarily involves the imbalance of blood sugar, which is mainly caused by abnormal metabolism of glucose. The pancreas secretes insulin into blood, so that the pathway conduction of insulin signals is caused, the body is accelerated to take in and utilize glucose, and after the glucose is taken by cells, glycolysis or complete oxidation and reduction are carried out to supply the energy requirement of the body or synthesize glycogen for standby. Once abnormality occurs in any one of the above processes, it causes imbalance of blood sugar and occurrence of insulin resistance, resulting in diabetes. The first occurrence of glucose entry into cells is the phosphorylation reaction catalyzed by Hexokinase (HK). 4 hexokinase isoenzymes are currently found in mammals, and among them, the liver cells exist in type IV, called Glucokinase (GK). Glucokinase is an inducing enzyme which has a high specificity for glucose and thus catalyzes a reaction of glucose to be an irreversible reaction. In addition to oxidation, glucose is unnecessarily synthesized into glycogen, and the phosphorylation of glycogen synthase is reduced due to inactivation or reduced expression of glycogen synthase kinase 3 (GSK 3 beta), so that the activity of glycogen synthase is increased, and the glycogen synthesis is increased.

In recent years, the understanding of the causes of development of diabetes has been focused on abnormalities in the intestinal flora, in addition to genetics, environment, immune inflammation, and the like. The intestinal flora enters our body from our birth, accompanies throughout life, and affects our life and death, so it is considered as "the second genome acquired in the human body. Therefore, once the intestinal flora of the organism is disordered due to some reasons, harmful substances are secreted by the intestinal flora into blood, and various tissue or systemic diseases can be caused, including senile dementia, diabetes and the like. And certain foods are frequently eaten, so that beneficial bacteria can be increased and harmful bacteria can be inhibited to a certain extent, and the disordered intestinal flora can be improved.

Currently, there are many chemical drugs on the market for the treatment of diabetes, such as: insulin, metformin, sulfonylureas, alpha-glucosidase inhibitors, and the like. Although the treatment effect is obvious, the toxic and side effects of hypoglycemia, allergy, ametropia and the like have to be noticed and prevented in the using process. The search for drugs or health care products that can reduce side effects has become a focus of attention and research.

Diabetes belongs to the category of 'diabetes' in traditional Chinese medicine, and is mainly caused by weak viscera, overeating fatness, fatigue, loss of tolerance, qi stagnation, blood stasis and the like. The traditional Chinese medicine is profound and profound, a plurality of traditional Chinese medicines are not only edible food materials which are usually eaten by people, but also medicinal materials which can treat diseases, and are called as medicine-food homology, and the traditional Chinese medicine mainly focuses on food therapy for treating chronic diseases.

Disclosure of Invention

The invention aims to provide a multifunctional food composition for assisting in reducing blood sugar, which is composed of medicinal and edible traditional Chinese medicinal materials, and a preparation method thereof. Has effects in lowering blood sugar, increasing insulin sensitivity, protecting liver, and improving intestinal flora.

In order to achieve the purpose, the invention adopts the following technical scheme: a multifunctional food composition for assisting in reducing blood sugar comprises the following components in parts by weight: 20-40 parts of Chinese yam, 10-30 parts of radix polygonati officinalis, 2-20 parts of medlar, 2-20 parts of white poria, 1-5 parts of bitter gourd, 1-5 parts of rhizoma polygonati, 1-5 parts of hawthorn, 1-5 parts of black sesame, 1-4 parts of mulberry leaf, 1-4 parts of dandelion, 1-2 parts of radix puerariae and 1-2 parts of platycodon grandiflorum.

Preferably, the food composition comprises the following components in parts by weight: 30 parts of Chinese yam, 20 parts of polygonatum, 10 parts of medlar, 10 parts of white poria, 3 parts of balsam pear, 3 parts of rhizoma polygonati, 3 parts of hawthorn, 3 parts of black sesame, 1 part of mulberry leaf, 1 part of dandelion, 1 part of kudzu root and 1 part of platycodon grandiflorum.

The multifunctional food composition for assisting in reducing blood sugar mainly comprises Chinese yam, polygonatum odoratum, medlar and chinaroot greenbrier rhizome.

Furthermore, the invention provides a preparation prepared by adding auxiliary materials into the health food composition, preferably, the health food composition can be prepared into any suitable oral preparation according to requirements, including tablets, capsules, powder, granules, effervescent agents or bagged steeping agents and the like, and the added auxiliary materials are pharmaceutically acceptable auxiliary materials.

The invention provides a preparation method of a multifunctional food composition for assisting in reducing blood sugar, which comprises the following steps: accurately weighing corresponding medicinal materials, cleaning the medicinal materials with clear water, steaming at 100 deg.C for 1 hr, oven drying at 65 deg.C, pulverizing, sieving with 120 mesh sieve, mixing the powders, and making into desired dosage form.

An application of a multifunctional food composition for assisting in reducing blood sugar in preparing a health-care product for assisting in reducing blood sugar and increasing insulin sensitivity.

A multifunctional food composition for assisting in lowering blood sugar is used for preparing liver protection health product, and can restore damaged liver lobule structure.

A multifunctional food composition for assisting in reducing blood sugar is used for preparing health products for improving intestinal dysbacteriosis, increasing the content of some beneficial bacteria, and reducing the content of some harmful bacteria.

The invention can be applied to different food products.

The efficacy activity of each raw material in the present invention is as follows.

Chinese yam: [ PROPERTIES AND FLAVOUR ] sweet and neutral. [ GUIJING ] can enter spleen, lung and kidney meridians. [ FUNCTIONS ] can tonify spleen and stomach, promote the production of body fluid and nourish lung, tonify kidney and arrest seminal emission. Can be used for treating spleen deficiency, anorexia, chronic diarrhea, lung deficiency, cough, asthma, spermatorrhea, leukorrhagia, frequent micturition, and diabetes due to deficiency heat. Tonifying spleen and stomach, benefiting kidney meridian; spleen tonifies and governs transportation and transformation, so it can tonify the disturbance of transportation and transformation and the disturbance of body fluid; the Chinese yam is sweet and slightly sour and can tonify blood of liver yin; entering kidney, slightly sour and astringent, and can be used for treating diseases due to kidney deficiency; it enters lung to astringe, so it can help lung to discharge turbid matter, and indirectly invigorate spleen and disperse body fluids.

Fragrant solomonseal rhizome: [ PROPERTIES AND FLAVOUR ] sweet and slightly cold. [ GUIJING ] enters lung and stomach meridians. [ FUNCTIONS ] can nourish yin, moisten dryness, promote the production of body fluid and quench thirst. Can be used for treating yin injury of lung and stomach, cough due to dryness-heat, dry throat, thirst, and diabetes due to internal heat.

Medlar: [ PROPERTIES AND FLAVOUR ] sweet and neutral. [ GUIJING ] enters liver and kidney meridians. [ FUNCTIONS ] can nourish liver and kidney, replenish vital essence and improve eyesight. Can be used for treating consumptive disease and essence deficiency, soreness of waist and knees, giddiness and tinnitus, internal heat, diabetes, blood deficiency, sallow complexion, blurred vision, kidney invigorating, essence replenishing, liver nourishing, and eyesight improving; the spleen is tonified, the transportation and transformation capacity is enhanced, and blood can be well enriched; it enters heart and heart governs blood vessels, so it can be used for treating blood vessel insecurity.

White poria: [ PROPERTIES AND FLAVOUR ] sweet, bland and mild. [ GUIJING ] can enter heart, lung, spleen and kidney meridians. [ FUNCTIONS ] can induce diuresis, remove dampness, invigorate spleen, and tranquilize mind. Can be used for treating edema, oliguria, phlegm and fluid retention, dizziness, palpitation, spleen deficiency, anorexia, loose stool, diarrhea, uneasiness, palpitation, insomnia, spleen invigorating, phlegm eliminating, heart calming, and tranquilizing; the spleen is tonified, nutrient substances are transported and dissolved into blood, and the blood is insufficient to help digestion; transporting and transforming water, eliminating phlegm-damp pathogen, and being neutral in nature and cold and heat.

Rhizoma polygonati: [ PROPERTIES AND FLAVOUR ] sweet and neutral. [ GUIJING ] can enter spleen, lung and kidney meridians. [ FUNCTIONS ] can invigorate qi, nourish yin, invigorate spleen, moisten lung, and tonify kidney. Can be used for treating weakness of spleen and stomach, asthenia, xerostomia, anorexia, lung deficiency, cough, essence and blood deficiency, and internal heat diabetes.

Hawthorn fruit: [ PROPERTIES AND FLAVOUR ] sour, sweet and slightly warm. [ GUIJING ] can enter spleen, stomach and liver meridians. [ FUNCTIONS ] can promote digestion, invigorate stomach, promote the circulation of qi, and remove blood stasis. Can be used for treating meat food stagnation, gastric distention, dysentery, abdominal pain, and dysentery.

Black sesame seed: [ PROPERTIES AND FLAVOUR ] sweet and neutral. [ GUIJING ] enters liver, kidney and large intestine meridians. [ FUNCTIONS ] can tonify liver and kidney, nourish essence and blood, moisten intestine and dry.

Mulberry leaf: [ PROPERTIES AND FLAVOURS ] sweet, bitter and cold. [ GUIJING ] enters lung and liver meridians. [ FUNCTIONS ] can dispel wind-heat, clear lung-heat, moisten dryness, clear liver-heat and improve eyesight.

Dandelion: [ PROPERTIES AND WESTERN ] is bitter, sweet and cold. [ GUIJING ] enters liver and stomach meridians. [ FUNCTIONS ] can clear away heat and toxic materials, relieve swelling, dissipate stagnation, induce diuresis, and treat stranguria.

Kudzu root: [ PROPERTIES AND FLAVOUR ] sweet, pungent and cool. [ GUIJING ] can enter spleen and stomach meridians. [ FUNCTIONS ] can relieve muscles and fever, promote salivation, promote eruption, elevate yang, and stop diarrhea. Can be used for treating thirst, diabetes, dysentery, and diarrhea.

Balloon flower: [ PROPERTIES AND WESTERN ] is bitter, pungent and mild. [ GUIJING ] enters lung meridian. [ FUNCTIONS ] can ventilate lung, relieve sore throat, dispel phlegm, and expel pus.

Compared with the prior art, the invention has the innovation point.

The health food which combines 12 traditional Chinese medicinal materials of Chinese yam, polygonatum, medlar, chinaroot greenbrier rhizome, balsam pear, rhizoma polygonati, hawthorn, black sesame, mulberry leaf, dandelion, kudzu root and platycodon root has no report on the synergistic effect in the aspects of reducing blood sugar and relieving diabetic complications, achieves the effect of assisting in reducing blood sugar, and simultaneously has the functions of increasing insulin sensitivity, protecting liver and improving intestinal flora. The 12 traditional Chinese medicinal materials are all selected from medicinal materials with homology of medicine and food in No. 2002 document of Duofan Law of Notification of the Ministry of health on further standardizing the management of health food raw materials, are natural and harmless, and are suitable for long-term administration. The whole body has sweet and mild flavor, does not generate cold and cool stimulation, and can be both considered when entering the channels and the five zang-organs and six fu-organs.

Besides the data of reference pharmacopoeia, ancient books, reported documents and the like, the invention also utilizes the bioinformatics analysis means of online databases of TCMSP, DAVID and the like to screen, enrich and analyze the components and targets of 4 Chinese medicaments of Chinese yam, polygonatum, medlar and white poria which are main components of the composition so as to support the reasonability and comprehensiveness of the formula.

The invention explains from multiple angles some mechanisms of the functions of assisting the reduction of blood sugar and the increase of insulin sensitivity, including the function of regulating the expression of related genes of insulin signal transduction and glucose metabolism and the improvement function of the disordered intestinal flora.

Drawings

FIG. 1 is a graph showing fasting blood glucose levels of each group in animal experiment 1 in example 8 of the present invention.

FIG. 2 is a graph showing blood glucose levels at 0.5 hours after glucose administration in each group of animal experiment 1 in example 8 of the present invention.

FIG. 3 is a graph showing blood glucose levels at 2 hours after glucose administration in each group of animal experiment 1 in example 8 of the present invention.

Fig. 4 is the area AUC under the 0, 0.5, 2 hour blood glucose curve after glucose administration for each group of animal experiment 1 of example 8 of the present invention.

In FIGS. 1-4: control represents a blank set, model represents a model set, SS-1 represents a model + example 1 set, SS-2 represents a model + example 2 set, SS-3 represents a model + example 3 set, SS-4 represents a model + example 4 set, SS-5 represents a model + example 5 set, SS-6 represents a model + example 6 set, and SS-7 represents a model + example 7 set.

FIG. 5 is a network diagram of the components-target points of 4 main drugs of Chinese yam, fragrant solomonseal rhizome, Chinese wolfberry and white poria in the invention. Where diamonds represent elements, circles represent targets, and black circles represent targets associated with diabetes.

FIG. 6 is a top-level bubble chart of the biological process or pathway ranking 15 related to diabetes, which is enriched by the 4 main drugs of Chinese yam, fragrant solomonseal rhizome, Chinese wolfberry and white poria according to the components and target points.

FIG. 7 shows fasting blood glucose values of each group in animal experiment 2 in example 12 of the present invention.

FIG. 8 is a graph showing blood glucose levels at 0.5 hour after glucose administration in each group of animal experiment 2 in example 12 of the present invention.

FIG. 9 is a graph showing blood glucose levels at 2 hours after glucose administration in each group of animal experiment 2 in example 12 of the present invention.

Fig. 10 is the area AUC under the 0, 0.5, 2 hour blood glucose curve after glucose administration for each group of animal experiment 2 of example 12 of the present invention.

FIG. 11 is the results of the insulin resistance index of each group in animal experiment 2 of example 12 of the present invention.

FIG. 12 shows the results of the liver index of each group in example 13 of the present invention.

FIG. 13 shows the renal index results of each group in example 13 of the present invention.

FIG. 14 is a graph of total protein levels in each group of sera in example 13 of the present invention.

FIG. 15 shows serum glutamate pyruvate transaminase levels of each group in example 13 of the present invention.

FIG. 16 is the serum creatinine levels of each group in example 13 of the present invention.

FIG. 17 shows serum uric acid levels of each group in example 13 of the present invention.

FIG. 18 shows HE-stained pathological sections of liver groups in example 14 of the present invention.

FIG. 19 is the expression level of hepatic Glucokinase (GK) mRNA of each group in example 15 of the present invention.

FIG. 20 shows the expression levels of mRNA for liver glycogen synthase kinase (GSK 3. beta.) in each group in example 15 of the present invention.

FIGS. 7-17, FIGS. 19-20: control represents blank group, normal represents normal administration high dose group, SS-L represents model + low dose group, SS-M represents model + medium dose group, SS-H represents model + high dose group, and positive represents positive control group.

FIG. 21 is a graph showing the results of intestinal flora in example 16 of the present invention.

Detailed Description

The statistical analysis result is processed by SPSS, and the statistical difference is considered to exist when P is less than or equal to 0.05.

First, screening test of the active ingredients of the present invention.

Example 1: weighing 20 parts of Chinese yam, 30 parts of polygonatum odoratum, 2 parts of medlar, 2 parts of white poria, 5 parts of bitter gourd, 5 parts of rhizoma polygonati, 5 parts of hawthorn, 5 parts of black sesame, 4 parts of mulberry leaf, 4 parts of dandelion, 2 parts of radix puerariae and 2 parts of platycodon grandiflorum, cleaning the medicinal materials with clear water, steaming at 100 ℃ for 1 hour, drying in a constant-temperature oven at 65 ℃, pulverizing by using a pulverizer, sieving by using a 120-mesh sieve, and uniformly mixing the powder.

Example 2: weighing 25 parts of Chinese yam, 25 parts of polygonatum odoratum, 8 parts of Chinese wolfberry, 8 parts of white poria, 3 parts of bitter gourd, 3 parts of rhizoma polygonati, 3 parts of hawthorn, 3 parts of black sesame, 3 parts of mulberry leaf, 3 parts of dandelion, 1 part of radix puerariae and 1 part of platycodon grandiflorum, and preparing the preparation method is the same as that of example 1.

Example 3: weighing 30 parts of Chinese yam, 20 parts of polygonatum odoratum, 10 parts of medlar, 10 parts of white poria, 3 parts of balsam pear, 3 parts of rhizoma polygonati, 3 parts of hawthorn, 3 parts of black sesame, 1 part of mulberry leaf, 1 part of dandelion, 1 part of radix puerariae and 1 part of platycodon grandiflorum, and preparing the preparation method is the same as that of example 1.

Example 4: weighing 40 parts of Chinese yam, 10 parts of polygonatum, 20 parts of medlar, 20 parts of white poria, 1 part of balsam pear, 1 part of rhizoma polygonati, 1 part of hawthorn, 1 part of black sesame, 2 parts of mulberry leaf, 2 parts of dandelion, 1 part of kudzu root and 1 part of platycodon root. The preparation method is the same as example 1.

Example 5: weighing 30 parts of gorgon fruit, 20 parts of polygonatum odoratum, 10 parts of medlar, 10 parts of white poria, 3 parts of balsam pear, 3 parts of rhizoma polygonati, 3 parts of hawthorn, 3 parts of black sesame, 1 part of mulberry leaf, 1 part of dandelion, 1 part of kudzu root and 1 part of platycodon root, and preparing the preparation method is the same as that of example 1.

Example 6: weighing 30 parts of Chinese yam, 20 parts of liquorice, 10 parts of medlar, 10 parts of white poria, 3 parts of balsam pear, 3 parts of rhizoma polygonati, 3 parts of hawthorn, 3 parts of black sesame, 1 part of mulberry leaf, 1 part of dandelion, 1 part of kudzu root and 1 part of platycodon grandiflorum, and preparing the preparation method is the same as that of example 1.

Example 7: weighing 30 parts of Chinese yam, 20 parts of polygonatum odoratum, 10 parts of mulberry, 10 parts of poria cocos, 3 parts of bitter gourd, 3 parts of rhizoma polygonati, 3 parts of hawthorn, 3 parts of black sesame, 1 part of mulberry leaf, 1 part of dandelion, 1 part of radix puerariae and 1 part of platycodon grandiflorum, and preparing the preparation method of the traditional Chinese medicine composition is the same as that of example 1.

Example 8: animal experiment 1.

Sample preparation: the health food compositions prepared in examples 1-7 were dispersed in water in a volume of 140mL, respectively.

Main reagents and instruments: a blood glucose meter.

Animals: male rats, 180. + -.20 g, of SPF grade were obtained from Liaoning Biotechnology Ltd.

8.1 grouping and administration.

(1) 90 healthy adult animals (male rats with SPF grade, 180 +/-20 g) are selected, the normal maintenance feed is adapted to be fed for 7 days, fasting blood glucose is measured and used as a basic value, and 10 healthy adult animals are randomly selected to be used as a blank group.

(2) The remaining 80 were then fasted for 24 hours (free drinking water) and then given a bolus of alloxan 150mg/kg injected intraperitoneally (immediately prior to use, in normal saline, over 30 minutes) to create a hyperglycemic model. After 5 days, fasting is carried out for 4 hours, tail blood is taken, blood sugar values are measured, and the blood sugar values are all larger than 10mmol/L, so that the molding is successful. The groups were randomly divided into 10 models, model + example 1, model + example 2, model + example 3, model + example 4, model + example 5, model + example 6, and model + example 7.

(3) The blank control group and the model control group are subjected to intragastric administration and are provided with the same volume of solvent, namely distilled water, with the volume of 1mL/100 g; the remaining groups were gavaged with the composition formulation of the corresponding example at 1mL/100g for 35 consecutive days.

(4) After the test was completed, all animals were fasted for 4 hours, and tail blood was taken to measure blood glucose and glucose tolerance.

8.2 sugar tolerance test.

The blank control group was not treated, the model control group was given the same volume of solvent, the other groups were given the corresponding dose of the composition, each group was given 2.5g/kg of glucose orally 20 minutes later, tail blood was taken, blood glucose levels at 0.5 and 2 hours after glucose administration were measured for each group, and the area AUC = (0 hour blood glucose +0.5 hour blood glucose) × 0.5/2+ (0.5 hour blood glucose +2 hour blood glucose) × 1.5/2 under the 0, 0.5, 2 blood glucose curve was calculated.

Example 9: the components and targets of the 4 main medicinal materials in the composition are screened, enriched and analyzed by applying a bioinformatics method.

(1) Inputting 4 Chinese medicinal materials into TCMSP (http:// lsp. nwu. edu. cn/tcmspsearch. php) database, respectively, and screening all the components according to pharmacokinetic parameters under the conditions that the oral bioavailability OB is more than or equal to 30%, the drug property DL is more than or equal to 0.18, or the oral bioavailability OB is more than or equal to 30% and the half-life HL is more than or equal to 4.

(2) And (4) corresponding the screened components to the target points in the database.

(3) The screened target points are input into a Uniprot (http:// www.uniprot.org /) database to standardize the target point names, corresponding to the gene names.

(4) Uploading the gene names of all targets to a DAVID (https:// DAVID. ncifcrf. gov/summary. jsp) database, carrying out GO-BP biological process and KEGG pathway enrichment analysis, and screening target genes related to the development of diabetes.

(5) And (3) performing component-target network construction on the selected components and targets by using Cytoscape3.2.1 software, and drawing a biological process or passage high-level bubble map related to diabetes.

Example 10: a low, medium and high dose composition according to the portion ratio in example 3 was prepared, i.e. example 3 was a low dose composition, with the medium dose composition having 5 times the content of each component of example 3 and the high dose composition having 10 times the content of each component of example 3, dispersed in a volume of 140mL of water, respectively.

Example 11: preparation of positive control:

weighing 3.90g of the capsule of the positive control medicament Tongrentang radix puerariae and Chinese yam, and uniformly dispersing in 140mL of clear water for later use.

Example 12: animal experiment 2.

Sample preparation: samples prepared in examples 10 and 11.

Main reagents and instruments: rat serum insulin ELISA kit, glucometer, full-automatic biochemical detector.

12.1 grouping and administration.

(1) 70 healthy adult animals (male rats with SPF grade, 180 +/-20 g) are selected, the normal maintenance feed is adapted to be fed for 7 days, fasting blood glucose is measured and used as a basic value, 20 healthy adult animals are randomly selected and respectively used as a blank group and a normal high-dose group (namely, a blank group and a normal high-dose group are not molded and are given with high dose), and 10 healthy adult animals are selected.

(2) The remaining 50 were then fasted for 24 hours (free drinking water) and then given a bolus of alloxan 150mg/kg injected intraperitoneally (immediately prior to use, in saline, over 30 minutes) to create a hyperglycemic model. After 5 days, fasting is carried out for 4 hours, tail blood is taken, blood sugar values are measured, and the blood sugar values are all larger than 10mmol/L, so that the molding is successful. The test samples were randomly divided into 10 model control groups, model + low dose group, model + medium dose group, model + high dose group, and positive control group.

(3) The blank control group and the model control group are subjected to intragastric administration and are provided with the same volume of solvent, namely distilled water, with the volume of 1mL/100 g; the remaining groups were gavaged with the corresponding composition formulation of the examples, 1mL/100g, for 35 consecutive days.

(4) After the test is finished, all animals are fasted for 4 hours, tail blood is taken to measure blood sugar and sugar tolerance, abdominal cavity venous blood is taken to centrifuge serum to measure serum insulin content, tissues such as liver, kidney and the like are picked up to measure weight, part of the same part of the liver is taken and stored in a refrigerator at minus 80 ℃ for standby, and part of the same part of the liver is stored in liquid with paraformaldehyde for standby.

12.2 sugar tolerance test: the experiment was as in example 8.2.

12.3 determination of insulin resistance index:

after fasting for 4 hours, blood of rats is taken, serum is centrifuged, and the content of serum insulin is detected by ELISA method.

(1) And (3) diluting the standard: and (3) taking 120 mu L of the standard substance, adding the equal volume of the standard substance diluent to serve as a No. 5 standard substance, taking 120 mu L of the No. 5 standard substance, adding the equal volume of the standard substance diluent to serve as a No. 4 standard substance, and diluting into the No. 3, No. 2 and No. 1 standard substances by analogy.

(2) The blank wells were not loaded with sample, biotin-labeled anti-INS antibody, streptavidin-HRP, but only with chromogenic A, B and stop solution, and the rest steps were performed in the same manner.

(3) The standard 50. mu.L, streptavidin-HRP 50. mu.L (standard already integrated with biotin antibody) was added to the wells.

(4) 40 mu L of sample is added into a sample hole to be detected, 10 mu L of anti-INS antibody and 50 mu L of streptavidin-HRP are added.

(5) After the addition, the sealing plate membrane is covered, the mixture is gently shaken and mixed, and the mixture is incubated at 37 ℃ for 60 minutes.

(6) Removing the sealing plate film, discarding the waste liquid, spin-drying, filling each hole with washing liquid, standing for 30 seconds, discarding, repeating the steps for 5 times, and patting to dry.

(7) The wells were first filled with 50. mu.L of developer A solution and then 50. mu.L of developer B solution (liquid B noted light shielding), incubated at 37 ℃ for 15 minutes in the absence of light.

(8) The reaction was stopped by adding 50. mu.L of a stop solution, and the absorbance OD value was measured in order at a wavelength of 450nm (measurement was completed within 10 minutes) with zero calibration in a blank well.

(9) From the OD values, a standard curve was drawn and the concentration of each insulin was calculated (standard curve OD = standard OD — blank OD, experimental group actual OD = experimental group OD — blank OD).

(10) The insulin resistance index ≈ fasting glucose x insulin/22.5 is calculated.

Example 13: and calculating the organ index and detecting the liver and kidney functions.

13.1 at the end of the experiment, harvested livers and kidneys were weighed and recorded, and the organ index = organ weight (g)/body weight of the experimental animal (g) × 100% was calculated.

13.2 full-automatic biochemical analyzer for detecting the contents of liver function index (total protein, alkaline phosphatase) and renal function index (creatinine and urea nitrogen) in serum.

Example 14: pathological section of liver.

14.1 wax block embedding.

(1) Washing with flowing water for 6 hours, and soaking with 70%, 80%, 90%, 95% and absolute ethyl alcohol gradient alcohol.

(2) Soaking in

xylene

1 and 2.

(3) And (6) wrapping a wax block.

(4) And (5) slicing by using a slicing machine.

14.2 HE staining.

(1) And handcuff the slices for 2 hours by a roasting machine.

(2)

Dewaxing xylene

1 and 2, and passing through absolute ethyl alcohol, 95 percent, 85 percent and 75 percent gradient alcohol.

(3) Tap water washing, hematoxylin staining and tap water washing.

(4) 1% hydrochloric acid alcohol, tap water washing, eosin dyeing and tap water washing.

(5) Gradient elution is carried out on 75%, 85%, 95% and absolute ethyl alcohol, and elution is carried out on xylene 1 and xylene 2.

(6) And sealing the neutral resin, airing and observing by a microscope.

Example 15: detection of the level of gene expression in the liver.

Main reagents and instruments: trizol reagent, DNA reverse transcription kit, DNA amplification kit, PCR amplification instrument, etc.

15.1 extraction of total RNA from liver tissue.

(1) The liver tissue frozen at-80 ℃ was removed, and approximately 50mg of the tissue was cut with a scalpel and placed in a test tube containing Trizol without RNase.

(2) The tissue was beaten into a homogenate by a homogenizer, mixed as uniformly as possible, and allowed to stand at room temperature for 5 minutes.

(3) Centrifuge at 12000g for 5 min at 4 ℃. Transferring the supernatant into a new centrifugal tube without RNA enzyme, adding chloroform according to the proportion of 5:1, violently shaking for about 30s, mixing uniformly, and standing for 5 minutes at room temperature.

(4) Centrifuge at 12000g for 15 min at 4 ℃. The supernatant liquid was transferred to a new rnase-free centrifuge tube.

(5) Adding isopropanol with the same volume, mixing by gently reversing the upper part and the lower part, and standing for 10 minutes at room temperature.

(6) Centrifuge at 12000g for 10 min at 4 ℃.

(7) The supernatant was carefully discarded, 1mL of 75% ethanol (DEPC water) pre-cooled at-20 ℃ was added to the pellet, and the pellet was washed by mixing the solution up and down.

(8) Centrifuge at 12000g for 5 min at 4 ℃.

(9) The supernatant was carefully discarded, air dried, the precipitate was dissolved with DEPC water and stored at-80 ℃.

15.2 reverse transcription of mRNA.

(1) mu.L of total RNA extracted from the liver was measured by an ultraminimetric UV analyzer and the concentration was recorded.

(2) Based on the measured concentration, the volume of RNA required was calculated as an amount of 1. mu.g of reverse transcription.

(3) Adding the calculated amount of template RNA and water into corresponding Reaction tubes, wherein each tube contains primer oligo (dT), gDNA Remover, Enzyme-Mix, 2 × Reaction-Mix, RNA and DEPC water.

(4) After fully and evenly mixed, the mixture is put into a PCR amplification instrument for reverse transcription.

(5) The complementary DNA (cDNA) is obtained and ready for use.

15.3 real-time fluorescent quantitative amplification (qPCR):

the selected internal reference gene is glyceraldehyde-3-phosphate dehydrogenase (GAPDH), the target gene is glucokinase, glycogen synthase kinase 3 beta, and the internal reference gene is synthesized in Shanghai.

(1) 12000g of primers are centrifuged for 15 minutes, and the corresponding Rase-free Water is added to each tube and diluted by 10 times for use.

(2) According to the experimental dosage, adding template cDNA, upstream and downstream primers, Master Mix and DEPC water to make up to 50 μ L per tube.

(3) The components are added into a 96-well plate with eight rows, and the 96-well plate is put into a Real Time PCR System instrument for amplification and Real-Time quantitative determination.

Example 16: and (4) detecting intestinal flora.

The detection groups are blank group, model + medium dose group.

16.1 MetaVx library construction and Illumina MiSeq sequencing.

(1) The concentration of the DNA sample was measured using a Qubit2.0 Fluorometer (Invitrogen, Carlsbad, Calif.), and a kit and sequencing library were constructed using a MetaVx-library.

(2) 30-50ng of DNA was used as a template, and 2 hypervariable regions including V3 and V4 were amplified in prokaryotic 16S rDNA using a series of PCR primers designed with Kinza-only.

(3) Library quality was checked using an Agilent 2100 bioanalyzer (Agilent Technologies, Palo Alto, CA, USA) and library concentration was determined by a qubit2.0 Fluorometer (Invitrogen, Carlsbad, CA).

16.2 the results are plotted as a heat map.

And (5) experimental results.

1. Example 8 of the present invention the fasting blood glucose levels and glucose tolerance results of each group of animal experiment 1 are shown in table 1 and fig. 1 to 4.

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Examples 5 to 7 are examples of the split of the equal division of example 3 of the present invention, and from the results, it can be seen that: the blood sugar of the model group is obviously increased, the fasting blood sugar of each group, the blood sugar of 0.5 hour and 2 hours after the glucose is given and the area under the blood sugar curve are obviously reduced compared with the model group, which shows that the composition has the effect of reducing the blood sugar, and the fasting blood sugar of each group, the blood sugar of 0.5 hour and 2 hours after the glucose is given and the area under the blood sugar curve of each group have no difference compared with the model group, which shows that the formula of the invention is reasonable and effective. The group of example 3 is the best group by combining the groups.

The TCMSP database analyzes the components and target spots of the 4 Chinese medicines, as shown in Table 2 and FIGS. 5-6.

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The 4 main raw materials of the traditional Chinese medicine are screened to obtain 103 potential active ingredients and 197 target points, wherein the total number of the enriched target points related to diabetes mellitus is 81, and the enriched target points account for more than 1/3 of the screened target points. The pathways of the first 15 are closely related to the metabolic processes of sugar and fat, and the rationality of the formula and the potential of the functional properties of assisting in reducing blood sugar and improving insulin sensitivity are fully explained from the aspect of big data bioinformatics analysis.

3. Example 12 results of fasting plasma glucose and glucose tolerance for each group of animal experiment 2 are shown in table 3 and figures 7-10.

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From the results it follows: compared with the blank group, the model group has obvious increase of the area AUC values under the fasting blood glucose value curve, the blood glucose value at 0.5 hour after the glucose administration, the blood glucose value at 2 hours after the glucose administration, the blood glucose value at 0 hour, 0.5 hour and 2 hours after the glucose administration, and the difference has statistical significance.

Compared with the model group, the model + low dose group, the model + medium dose group, the model + high dose group and the positive control group, the fasting blood glucose value, the blood glucose value 0.5 hour after the glucose administration, the blood glucose value 2 hour after the glucose administration, the AUC values of the areas under the blood glucose value curves 0, 0.5 and 2 hours after the glucose administration are obviously reduced, and the difference has statistical significance. The hypoglycemic effect of the invention has a relationship with the dosage, wherein the medium dosage is more than the high dosage and more than the low dosage.

The blank + high dose group, i.e., the high dose group administered to normal animals, showed no statistical difference in the area AUC under the fasting blood glucose level curve, the blood glucose level at 0.5 hour after administration of glucose, the blood glucose level at 2 hours after administration of glucose, and the blood glucose level at 0, 0.5, and 2 hours after administration of glucose, compared to the blank group.

4. The results of the calculation of the insulin resistance index for each group are shown in table 4 and fig. 11.

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From the results it follows: compared with the blank group, the model group has obvious increase of the insulin resistance index value, and the difference has statistical significance.

Compared with the model group, the model + low dose group, the model + medium dose group, the model + high dose group and the positive control group, the insulin resistance index value is obviously reduced, and the difference has statistical significance. The effect of the invention on increasing insulin sensitivity is related to the dosage, wherein the dosage is more than high dosage and less than low dosage.

The blank + high dose group, i.e., the high dose group given to normal animals, did not statistically differ from the blank group in insulin resistance index values.

5. The organ index results of example 13 of the present invention are shown in Table 5 and FIGS. 12 to 13.

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From the results it follows: the model group showed an increase in visceral index compared to the blank group, but did not show statistical differences.

Compared with the model group, the model + low dose group, the model + medium dose group, the model + high dose group and the positive control group, the index value of the visceral organs is reduced, but the statistical difference is not generated.

The group of blank + high dose, i.e., the group of high dose given to normal animals, had no statistical difference in organ index values compared to the blank group.

6 liver and kidney function index results of example 13 of the present invention are shown in Table 6 and FIGS. 14 to 17.

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From the results it follows: the serum total protein and creatinine levels of the model group are increased compared with the blank group, but the serum total protein and creatinine levels are not statistically different; the serum total protein and creatinine levels were reduced without statistical difference compared to the model group for the model + low dose group, the model + medium dose group, the model + high dose group, and the positive group.

The serum glutamic-pyruvic transaminase and uric acid levels of the model group are obviously increased compared with those of the blank group, and the statistical difference is realized; the level of serum glutamic pyruvic transaminase and uric acid is obviously reduced compared with the model group and the model group, and the model group and the low-dose group, the model group and the medium-dose group, the model group and the high-dose group and the positive group have statistical significance.

The blank + high dose group, i.e., the high dose group given to normal animals, showed no difference in serum total protein, creatinine, glutamic-pyruvic transaminase, uric acid levels, compared to the blank group.

7 liver pathology HE staining results for each group are shown in fig. 18.

From the results it follows: comparing hepatic lobules of liver tissues of the model group with the blank group, the structure of hepatic cord is changed, the arrangement of cells is irregular, and the central venous area is reduced; the model + medium dose group can basically recover the disordered lobular structure, approaches the blank group and is better than the change of the positive control group; the hepatic lobular structural damage of the model + low dose, high dose group was also improved; the blank + high dose group (i.e., the high dose group given to normal animals) did not undergo pathological changes compared to the blank group.

8 results of the mRNA expression levels of the liver genes of each group are shown in Table 7, FIGS. 19 to 20.

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From the results it follows: compared with the blank group, the mRNA level of the hepatic Glucokinase (GK) is obviously reduced, and the difference has statistical significance; the positive control group is compared with the model group, the mRNA level of the hepatic Glucokinase (GK) is obviously increased, and the difference has statistical significance. The blank + high dose group, i.e., the high dose group given to normal animals, had no statistical difference in the mRNA level of hepatic Glucokinase (GK) compared to the blank group.

Compared with the blank group, the mRNA level of liver glycogen synthase kinase 3 beta (GSK 3 beta) is obviously increased, and the difference has statistical significance; the mRNA level of liver glycogen synthase kinase 3 beta (GSK 3 beta) is obviously reduced and the difference has statistical significance when the model + low dose group, the model + medium dose group and the model + high dose group are compared with the positive control group and the model group.

The blank + high dose group, i.e., the high dose group given to normal animals, showed no statistical difference in the mRNA level of liver glycogen synthase kinase 3 β (GSK 3 β) compared to the blank group.

Results of intestinal flora are shown in table 8 and fig. 21.

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From the results it follows: results for intestinal flora show: some bacteria such as lachnospirillum which are beneficial to the human body are obviously reduced in the model group, and the content of the bacteria is increased; some bacteria which are unfavorable for human body, such as pasteurella, fusobacterium, proteus, streptococcus, are increased in the model group, and the invention can reduce the increased flora, which shows that the invention can improve the function of the disordered intestinal flora.

To summarize: animal experiments show that compared with a model group, the fasting blood glucose value, the blood glucose values at 0.5 and 2 hours after glucose administration in a glucose tolerance experiment and the areas under the blood glucose curves at 0, 0.5 and 2 hours are obviously reduced, which shows that the invention has the function of assisting in reducing blood glucose; the insulin resistance index of the invention shows a descending trend compared with the model group, and has statistical difference, which shows that the invention has the function of insulin sensitization; the blank + high dose group, that is, the high dose group of the health food given by the normal animal, has no obvious change in the blood sugar values at 0, 0.5 and 2 hours, the areas under the blood sugar curves at 0, 0.5 and 2 hours and the insulin resistance index, which shows that the health food can not generate blood sugar fluctuation or insulin resistance when being taken by the normal group, and the health food can be eaten safely.

The change of liver and kidney index, the detection of serum liver and kidney function index and the HE staining result of liver pathological section show that the invention has certain liver and kidney protection function and certain recovery function to the pathological damage of diabetic liver.

The detection of the mRNA level of the gene in the liver proves that the invention can regulate the expression of the genes of glucokinase and glycogen synthase kinase 3, is beneficial to the reduction of blood sugar, and indicates that the health food can regulate the expression of related genes in the processes of insulin signal transduction and glucose metabolism.

The results of the intestinal flora show that the invention can increase the content of probiotics and reduce the flora which is unfavorable to human body, thereby improving the disordered intestinal flora and being beneficial to the health of the organism.

The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. The multifunctional food composition for assisting in reducing blood sugar is characterized by being prepared from the following components in parts by weight: 30 parts of Chinese yam, 20 parts of polygonatum, 10 parts of medlar, 10 parts of white poria, 3 parts of balsam pear, 3 parts of rhizoma polygonati, 3 parts of hawthorn, 3 parts of black sesame, 1 part of mulberry leaf, 1 part of dandelion, 1 part of kudzu root and 1 part of platycodon grandiflorum.

2. The multifunctional food composition for assisting in reducing blood sugar according to claim 1, wherein the food composition is prepared into an oral preparation comprising a tablet, a capsule, a powder, a granule, an effervescent or a bag infusion by adding an auxiliary material, wherein the auxiliary material is generally a pharmaceutically acceptable auxiliary material.

3. The preparation method of the multifunctional food composition for assisting in lowering blood sugar of claim 1, comprising the steps of: accurately weighing corresponding medicinal materials, cleaning the medicinal materials with clear water, steaming at 100 deg.C for 1 hr, oven drying at 65 deg.C, pulverizing, sieving with 120 mesh sieve, mixing the powders, and making into desired dosage form.

4. The use of the multifunctional food composition for assisting in lowering blood sugar according to claim 1 in the preparation of health products for assisting in lowering blood sugar, increasing insulin sensitivity, protecting liver, and improving intestinal flora.

5. Use of the multi-functional food composition for assisting in lowering blood glucose according to claim 1 in the preparation of a food.