A blood sugar lowering composition and its preparation method and application

Abstract

The invention discloses a hypoglycemic composition and a preparation method and application thereof, belonging to the technical field of hypoglycemic drugs. The hypoglycemic composition comprises raw materials in parts by weight: 15-20 parts of pine mushroom polysaccharide, 10-15 parts of schisandra extract, 3-5 parts of lotus root extract, 1-2 parts of lotus leaf extract, 1-3 parts of lotus seed embryo extract, 8-10 parts of sargassum polysaccharide, 8-10 parts of cordyceps polysaccharide, 5-8 parts of tuckahoe polysaccharide, 2-5 parts of ganoderma lucidum polysaccharide, and 3-5 parts of sargassum polysaccharide. The hypoglycemic composition can improve the hypoglycemic effect of pine mushroom polysaccharide from different action pathways, and hypoglycemia is more efficient, and can repair pancreatic islet β cells and improve the hypoglycemic effect of type I diabetes.

Description

Technical Field

[0001] The present invention relates to the technical field of blood sugar lowering drugs, and in particular to a blood sugar lowering composition, a preparation method and application thereof. Background Art

[0002] Diabetes is a metabolic disease characterized by high blood sugar. With the improvement of living standards, people's lifestyles and dietary structures have also changed. In recent years, diabetes has seriously endangered human health. With high prevalence and low treatment rate, diabetes has become one of the three major fatal diseases in humans, and its mortality rate is second only to cardiovascular and cerebrovascular diseases and cancer. Type 1 diabetes and type 2 diabetes are the two main categories of diabetes. Type 1 diabetes refers to the inability of the patient's body to produce insulin normally. Oral hypoglycemic drugs alone cannot lower blood sugar. Patients need to take insulin for treatment, but long-term use of these drugs will have inevitable toxic side effects on the human body, easily leading to adverse reactions such as hypoglycemia, vomiting and diarrhea, which also limits its application. Type 2 diabetes is often referred to as non-insulin-dependent diabetes. It can be treated by diet control, drugs, appropriate physical exercise, etc., and does not necessarily require insulin injection. If the body maintains a high blood sugar state for a long time, it will cause the occurrence of a variety of diabetic complications and harm human health. Therefore, it is of great significance to find drugs with no side effects and good blood sugar lowering effects.

[0003] Fungal polysaccharides in edible fungi have a variety of medicinal and health-care effects. A large number of studies have shown that edible fungi polysaccharides can lower blood lipids, lower blood sugar, protect the liver and promote bile secretion, and enhance human immunity. Polysaccharides are the main biologically active substances in edible fungi, and their biggest advantages are low toxicity and few side effects. Therefore, the development of medicines and health products made of edible fungi polysaccharides has become one of the hot research areas in the deep processing of edible fungi. There are three precious active substances in Matsutake, double-chain Matsutake polysaccharides, Matsutake polypeptides and Matsutake alcohol. Matsutake polysaccharides are sugar chains bound by glycosidic bonds. They are polymerized sugars composed of at least 10 monosaccharides and are high-molecular carbohydrates. Matsutake polysaccharides are not a pure substance, but a mixture of polysaccharides with different degrees of polymerization. Matsutake polysaccharides have the effect of lowering blood sugar and can effectively improve the symptoms of diabetic patients. Modern medicine shows that Matsutake has the effect of treating diabetes. Main modes of action: 1. Increase insulin content in the body: Matsutake can stimulate pancreatic β cells to secrete insulin, protect and repair pancreatic β cells, and increase insulin content in the body; 2. Reduce blood sugar content after meals: α-glucosidase inhibitor is a new type of hypoglycemic drug. Experiments have shown that α-glucosidase inhibitor (GI) extracted from Matsutake can reduce blood sugar after meals in diabetic patients; 3. Direct hypoglycemia: Matsutake can improve insulin resistance, accelerate liver glucose metabolism, and has a direct hypoglycemic effect. This application proposes a hypoglycemic composition containing Matsutake polysaccharide. Summary of the invention

[0004] As described in the above-mentioned prior art, one of the purposes of the present invention is to provide a hypoglycemic composition containing Matsutake extract, which can improve the hypoglycemic effect of Matsutake polysaccharide from different pathways of action, lower blood sugar more efficiently, and can repair pancreatic β cells, thereby improving the hypoglycemic effect of type 1 diabetes.

[0005] The second object of the present invention is to provide a method for preparing the hypoglycemic composition, which has simple steps and can be used for industrial production of the hypoglycemic composition.

[0006] The third object of the present invention is to provide the use of the hypoglycemic composition in the preparation of a hypoglycemic drug.

[0007] One of the purposes of the present invention is achieved by the following technical solution:

[0008] A blood sugar lowering composition comprises raw materials in parts by weight: 15-20 parts of matsutake polysaccharide, 10-15 parts of schisandra extract, 3-5 parts of lotus root extract, 1-2 parts of lotus leaf extract, 1-3 parts of lotus embryo extract, 8-10 parts of sargassum polysaccharide, 8-10 parts of cordyceps polysaccharide, 5-8 parts of tuckahoe polysaccharide, 2-5 parts of ganoderma polysaccharide and 3-5 parts of sargassum polysaccharide.

[0009] Schisandra extract: Schisandra glycosides are the active ingredient. Schisandra glycosides can inhibit sodium-glucose co-transporter 2 (SGLT2), promote the regeneration of pancreatic β cells in diabetic mice, and reveal the specific path of regeneration: 1. Promote β cell proliferation and increase the number of β cells; 2. Activate pancreatic precursor cells (also known as pancreatic stem cells) and promote their differentiation to form new β cells, producing more β cells from the source; 3. Induce other types of pancreatic cells (such as α cells that secrete glucagon) to transform into β cells that secrete insulin.

[0010] Sargassum polysaccharide can significantly increase glucose consumption, glycogen synthesis, and the activities of pyruvate kinase (PK) and hexokinase (HK) in insulin-resistant HepG2 cells. The mechanism is to improve insulin resistance by upregulating the expression levels of insulin receptor substrate-1 (IRS-1), glycogen synthase (GS), phosphoinositide 3-kinase (PI3K) and glucose transporter 4 (GLUT4).

[0011] Cordyceps sinensis polysaccharides inhibit the colon Toll-like receptor 4 (TLR4) / nuclear transcription factor (NF-κB) pathway and upregulate the expression of intestinal tight junction proteins, thereby improving glucose and serum lipid metabolism, hormone secretion and complications, reducing the permeability of intestinal epithelial cells in diabetic mice, slowing down the gastrointestinal absorption of sugar, and improving insulin sensitivity, thereby achieving the effect of lowering blood sugar.

[0012] Poria polysaccharide can change the microbial community structure in the intestines of ob / ob diabetic mice, increase the increase of butyrate-producing species of Lachnospiraceae and Carboxybacterium, increase the butyrate content in the intestines of ob / ob diabetic mice, improve intestinal mucosal damage, protect the integrity of the intestinal mucosa, reduce the proportion of Firmicutes and Bacteroidetes, increase the abundance of polysaccharide-utilizing bacteria, and promote sugar metabolism in the body.

[0013] Ganoderma lucidum polysaccharides promote insulin-stimulated glucose uptake, inhibit the expression of protein tyrosine phosphatase 1B (PTP1B) at the mRNA level, phosphorylate insulin receptor substrate-1 (IRS1), and activate phosphatidylinositol 3-kinase (PI3K) and protein kinase B (Akt), and also phosphorylate adenosine monophosphate-activated protein kinase (AMPK), thereby upregulating the expression of glucose transporter 4 (GLUT4).

[0014] Sargassum fusiformis polysaccharide increases the expression of lipid droplet coating protein gene, which can improve lipid metabolism disorders and reduce insulin resistance.

[0015] Furthermore, the raw materials in parts by weight include 20 parts of Matsutake polysaccharide, 10 parts of Schisandra chinensis extract, 4 parts of lotus root extract, 1 part of lotus leaf extract, 3 parts of lotus seed embryo extract, 10 parts of Sargassum polysaccharide, 8 parts of Cordyceps sinensis polysaccharide, 5 parts of Poria polysaccharide, 3 parts of Ganoderma lucidum polysaccharide, and 4 parts of Sargassum fusiforme polysaccharide.

[0016] Furthermore, the preparation method of Tricholoma matsutake polysaccharide comprises the following steps:

[0017] S1. Cultivating Tricholoma matsutake mycelia by liquid submerged fermentation, transferring the Tricholoma matsutake mycelia to a solid culture medium, and culturing Tricholoma matsutake mycelia and Tricholoma matsutake fruiting bodies by solid fermentation;

[0018] S2, grinding the cultured mycelium of Tricholoma matsutake and the cultured fruiting body of Agaricus blazei, adding water to the ground materials, adjusting the pH of the solution to between 4.0 and 5.5, adjusting the temperature of the solution to between 50 and 60° C., adding pectinase and cellulase to the solution, performing ultrasound-assisted extraction during the enzymolysis process, and obtaining an enzymolysis solution after the enzymolysis is completed;

[0019] S3, adding ammonium sulfate to the enzymatic hydrolyzate for salting out and precipitation to obtain a precipitate and a supernatant;

[0020] S4, adding anhydrous ethanol to the supernatant to obtain a crude product of Tricholoma matsutake polysaccharide;

[0021] S5. Decolorizing the crude Tricholoma matsutake polysaccharide with a macroporous resin, separating with the dextran gel column chromatography, and collecting components in a required molecular weight range to obtain the Tricholoma matsutake polysaccharide.

[0022] The second object of the present invention is achieved by adopting the following technical solution:

[0023] A method for preparing a blood sugar-lowering composition comprises the following steps:

[0024] The Tricholoma matsutake polysaccharide, Schisandra chinensis extract, lotus root extract, lotus leaf extract, lotus seed embryo extract, Sargassum polysaccharide, Cordyceps militaris polysaccharide, Cordyceps sinensis polysaccharide, Poria cocos polysaccharide, Ganoderma lucidum polysaccharide and Sargassum fusiformis polysaccharide are weighed according to the formula amount and dissolved in 70% (V / V) ethanol aqueous solution in sequence, and freeze-dried to obtain the blood sugar lowering composition.

[0025] The third object of the present invention is to provide a hypoglycemic composition for use in the preparation of a hypoglycemic drug.

[0026] Furthermore, the hypoglycemic composition can also be used as an active ingredient and a pharmaceutically acceptable excipient to prepare the hypoglycemic drug; the excipient is one of an excipient, a disintegrant, a sweetener, a binder, a coating agent and a bulking agent.

[0027] Furthermore, the blood sugar lowering drug is one of a solution, an emulsion, a suspension, an injection, a powder, a granule, a tablet, and a pill.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] (1) The present invention provides a hypoglycemic composition, wherein Matsutake polysaccharide can repair pancreatic β cells, and Schisandra chinensis extract and Matsutake polysaccharide synergistically promote the repair of pancreatic β cells, thereby enhancing the therapeutic effect on type I diabetes; Sargassum polysaccharide can significantly improve the glucose consumption, glycogen synthesis, and the activities of pyruvate kinase (PK) and hexokinase (HK) in insulin-resistant HepG2 cells, and synergizes with Ganoderma lucidum polysaccharide to improve insulin resistance, promote the sensitivity of liver cells to the effects of insulin, and enhance the hypoglycemic effect of type II diabetes; Cordyceps sinensis polysaccharide inhibits the colon Toll-like receptor 4 (TLR4) / nuclear transcription factor (NF-κB) pathway and upregulates the expression of intestinal tight junction proteins, thereby reducing the permeability of intestinal epithelial cells in diabetic mice, slowing down the gastrointestinal absorption of sugar, and improving insulin sensitivity, thereby achieving the effect of lowering blood sugar; Poria cocos polysaccharide can also improve the microbial community structure in the intestine of diabetic mice and enhance the activity of polysaccharide-utilizing bacteria. The content of polyphenols in lotus root pulp, lotus root skin, lotus root nodes, lotus seeds and lotus leaves is very rich, and the content of alkaloids in lotus seeds is high. Polyphenols can lower blood lipid levels by inhibiting liver lipogenesis factors such as PPAR-γ, GLUT4 and leptin. Alkaloids can inhibit lipase, causing lipase to lose its decomposition ability and restrict fat from entering the blood, thereby producing a lipid-lowering effect, improving the insulin resistance problem caused by obesity and making it easier to lower blood sugar. Sargassum fusiformis polysaccharide increases the expression of lipid droplet coating protein genes, improves lipid metabolism disorders and reduces insulin resistance, and further improves and promotes lipid metabolism. Therefore, the above ingredients can improve the blood sugar-lowering effect of pine mushroom polysaccharide from different pathways of action, making blood sugar lowering more efficient, and synergize with pine mushroom polysaccharide to play a blood sugar-lowering role and improve the blood sugar-lowering effect.

[0030] (2) The preparation of the Matsutake extract of the present invention is through liquid deep fermentation technology, a large amount of Matsutake mycelium is obtained, and the target active substance is extracted from it, so as to realize the expanded cultivation of Matsutake, which is of great significance to make up for the shortage of wild resources. Compared with traditional artificial cultivation technology, liquid deep fermentation has the advantages of short time, easy industrial production, cost saving, stable yield and easy quality control. Therefore, improving its fermentation process and culture conditions plays an important role. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] In order to more clearly illustrate the embodiments of the present invention, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying any creative work.

[0032] Figure 1This is a graph showing the results of measuring glucose consumption in insulin resistant (IR) HepG2 cells provided in Experimental Example 1;

[0033] Figure 2 The results of the hexokinase activity assay of insulin resistance (IR) HepG2 provided in Experimental Example 1;

[0034] Figure 3 The results of the pyruvate kinase activity assay of insulin resistant (IR) HepG2 cells provided in Experimental Example 1;

[0035] Figure 4 This is the result of measuring the glycogen content in insulin resistant (IR) HepG2 cells provided in Experimental Example 1. DETAILED DESCRIPTION

[0036] The present invention is further described below in conjunction with specific embodiments. It should be noted that, under the premise of no conflict, the embodiments or technical features described below can be arbitrarily combined to form a new embodiment.

[0037] Schisandra chinensis extract was purchased from Changsha Shanghe Biotechnology Co., Ltd.;

[0038] Sargassum polysaccharide was purchased from Xi'an Pris Bioengineering Co., Ltd.;

[0039] Cordyceps sinensis polysaccharide was purchased from Chengdu Pusi Biotechnology Co., Ltd.;

[0040] Poria polysaccharide was purchased from Shanghai Moqi Biotechnology Co., Ltd.;

[0041] Ganoderma lucidum polysaccharide was purchased from Shanghai Ronghe Pharmaceutical Technology Development Co., Ltd.;

[0042] Glycogen content, hexokinase (HK) activity, and pyruvate kinase (PK) activity detection kits were purchased from Shanghai Yansheng Biochemical Reagent Co., Ltd.;

[0043] Sargassum fusiformis polysaccharide was provided by the Key Laboratory of Mudanjiang Medical College.

[0044] Preparation of lotus root extract: 100 g of dry lotus root was refluxed with 70% (V / V) ethanol aqueous solution for 12 h, and after cooling, it was filtered through filter paper with a pore size of 1.2 μm. The filtered lotus root extract was concentrated under reduced pressure below 50°C and completely dried in a vacuum dryer to obtain 4.5 g of lotus root extract.

[0045] Preparation of lotus seed extract: 100 g of lotus seeds were refluxed with a 70% (V / V) ethanol aqueous solution for 12 h, cooled, and filtered through filter paper with a pore size of 1.2 μm. The filtered lotus seed extract was concentrated under reduced pressure below 50° C. and completely dried using a reduced pressure dryer to obtain 11.12 g of lotus extract.

[0046] Preparation of lotus leaf extract: 100 g of dried lotus leaves were heated to reflux with a 70% (V / V) ethanol aqueous solution for 12 h, cooled and filtered through filter paper with a pore size of 1.2 μm. The filtered lotus leaf extract was concentrated under reduced pressure below 50°C and completely dried with a reduced pressure dryer to obtain 7.54 g of lotus leaf extract.

[0047] Example 1

[0048] The present embodiment provides a method for preparing Tricholoma matsutake polysaccharide, comprising the following steps: S1, culturing Tricholoma matsutake mycelium by liquid submerged fermentation, transferring the Tricholoma matsutake mycelium to a solid culture medium, and culturing the Tricholoma matsutake mycelium and Tricholoma matsutake fruiting bodies by solid fermentation;

[0049] Specifically, the liquid deep fermentation medium is: 2% corn starch, 4% soybean powder, 0.002% vitamin B6, and the balance is water; 35L of the medium is prepared according to the above formula, poured into a 50L fermentation tank, and sterilized at 121°C for 30 minutes. The conditions for culturing selenium-enriched pine mushroom mycelium using liquid deep fermentation are as follows:

[0050] The fermentation process is as follows:

[0051] The slant strain was transferred to a triangular flask filled with seed culture medium, and cultured at 25-28°C with shaking for 7-10 days to obtain the cultured strain;

[0052] The cultured bacteria were inoculated into a liquid submerged fermentation medium at an inoculum rate of 10%, and the fermentation parameters were a temperature of 23°C, a dissolved oxygen of 20-80%, a natural pH, and the culture was continued for 6 days before the fermentation was terminated;

[0053] S2, grinding the cultured mycelium of Tricholoma matsutake and the cultured fruiting body of Agaricus blazei, adding water to the ground materials, adjusting the pH of the solution to between 4.0 and 5.5, adjusting the temperature of the solution to between 50 and 60° C., adding pectinase and cellulase to the solution, performing ultrasound-assisted extraction during the enzymolysis process, and obtaining an enzymolysis solution after the enzymolysis is completed;

[0054] Specifically, in step S2, the amount of pectinase and cellulase added to the solution is 2.5% by mass;

[0055] S3, adding ammonium sulfate to the enzymatic hydrolyzate for salting out and precipitation to obtain a precipitate and a supernatant;

[0056] Specifically, in the salting out step, the amount of ammonium sulfate added is such that the ammonium sulfate saturation of the solution is 75-85%, and the precipitation is carried out for 12-36 hours; the dialysis is carried out for 8-12 hours using a dialysis bag with a molecular weight cut-off of 3.5 kD;

[0057] The above-mentioned ammonium sulfate saturation refers to the saturation of the saturated ammonium sulfate solution as 100%, and the ammonium sulfate saturation of 75-85% means that the ammonium sulfate content is 75-85% of the ammonium sulfate in the saturated ammonium sulfate solution. Controlling the ammonium sulfate saturation of the solution within the above range has a better precipitation effect. Dialysis in the above manner can remove salt and small molecular impurities in the precipitate and maximize the retention of active ingredients;

[0058] S4. Add anhydrous ethanol to the supernatant to obtain the crude pine mushroom polysaccharide. The supernatant is pine mushroom alcohol, which is reserved for later use.

[0059] S5. Decolorize the crude pine mushroom polysaccharide with a macroporous resin, separate it with a gel column chromatography, collect the components in the required molecular weight range, sterilize and package it, and then obtain pine mushroom polysaccharide.

[0060] Example 2

[0061] The present embodiment provides a blood sugar lowering composition, which includes, by weight, 20 parts of Matsutake polysaccharide, 10 parts of Schisandra chinensis extract, 4 parts of lotus root extract, 1 part of lotus leaf extract, 3 parts of lotus seed embryo extract, 10 parts of Sargassum polysaccharide, 8 parts of Cordyceps polysaccharide, 5 parts of Poria polysaccharide, 3 parts of Ganoderma lucidum polysaccharide, and 4 parts of Sargassum fusiformis polysaccharide.

[0062] The preparation method of a hypoglycemic composition provided in this embodiment comprises the following steps: weighing matsutake polysaccharide, lotus root extract, lotus leaf extract, lotus seed embryo extract, sargassum polysaccharide, cordyceps polysaccharide, tuckahoe polysaccharide, ganoderma lucidum polysaccharide, and fusiforme polysaccharide according to the formula amount, dissolving them in 70% (V / V) ethanol aqueous solution in turn, and freeze-drying to obtain a hypoglycemic composition.

[0063] (3) The inhibitory effect of Tricholoma matsutake polysaccharide on α-amylase is non-competitive and in a dose-dependent manner; Tricholoma matsutake polysaccharide has a mixed inhibitory effect on α-glucosidase and in a dose-dependent manner. Insulin resistance (IR) can lead to a decrease in the levels of hexokinase (HK) and pyruvate kinase (PK). Tricholoma matsutake polysaccharide can restore the levels of hexokinase (HK) and pyruvate kinase (PK) and promote glucose metabolism.

[0064] Example 3

[0065] The present embodiment provides a blood sugar lowering composition, which includes, by weight, 18 parts of Matsutake polysaccharide, 12 parts of Schisandra chinensis extract, 5 parts of lotus root extract, 2 parts of lotus leaf extract, 1 part of lotus seed embryo extract, 8 parts of Sargassum polysaccharide, 9 parts of Cordyceps polysaccharide, 8 parts of Poria polysaccharide, 5 parts of Ganoderma lucidum polysaccharide, and 3 parts of Sargassum fusiformis polysaccharide.

[0066] The preparation method of a hypoglycemic composition provided in this embodiment comprises the following steps: weighing matsutake polysaccharide, lotus root extract, lotus leaf extract, lotus seed embryo extract, sargassum polysaccharide, cordyceps polysaccharide, tuckahoe polysaccharide, ganoderma lucidum polysaccharide, and fusiforme polysaccharide according to the formula amount, dissolving them in 70% (V / V) ethanol aqueous solution in turn, and freeze-drying to obtain a hypoglycemic composition.

[0067] Example 4

[0068] The present embodiment provides a blood sugar lowering composition, which includes, by weight, 15 parts of Matsutake polysaccharide, 15 parts of Schisandra chinensis extract, 3 parts of lotus root extract, 1 part of lotus leaf extract, 2 parts of lotus seed embryo extract, 9 parts of Sargassum polysaccharide, 10 parts of Cordyceps polysaccharide, 7 parts of Poria polysaccharide, 2 parts of Ganoderma lucidum polysaccharide, and 5 parts of Sargassum fusiformis polysaccharide.

[0069] The preparation method of a hypoglycemic composition provided in this embodiment comprises the following steps: weighing matsutake polysaccharide, lotus root extract, lotus leaf extract, lotus seed embryo extract, sargassum polysaccharide, cordyceps polysaccharide, tuckahoe polysaccharide, ganoderma lucidum polysaccharide, and fusiforme polysaccharide according to the formula amount, dissolving them in 70% (V / V) ethanol aqueous solution in turn, and freeze-drying to obtain a hypoglycemic composition.

[0070] Example 5

[0071] This embodiment provides a hypoglycemic drug. According to different pharmaceutical excipients, a hypoglycemic composition can also be used as an active ingredient and a pharmaceutically acceptable pharmaceutical excipient to prepare a hypoglycemic drug.

[0072] The pharmaceutical excipient may be one of an excipient, a disintegrant, a sweetener, a binder, a coating agent and a bulking agent.

[0073] Depending on the different drug excipients, the blood sugar-lowering drug can be a solution, emulsion, suspension, injection, powder, granule, tablet, or pill.

[0074] Comparative Example 1

[0075] The difference from Example 2 is that the blood sugar lowering composition provided in Comparative Example 1 includes, in terms of weight parts, 30 parts of Matsutake polysaccharide, 4 parts of lotus root extract, 1 part of lotus leaf extract, 3 parts of lotus embryo extract, 10 parts of Sargassum polysaccharide, 8 parts of Cordyceps polysaccharide, 5 parts of Poria polysaccharide, 3 parts of Ganoderma lucidum polysaccharide, and 4 parts of Sargassum fusiforme polysaccharide.

[0076] Comparative Example 2

[0077] The difference from Example 2 is that the blood sugar lowering composition provided in Comparative Example 1 includes, in terms of weight parts, 20 parts of Matsutake polysaccharide, 10 parts of Schisandra chinensis extract, 4 parts of lotus root extract, 1 part of lotus leaf extract, 3 parts of lotus seed embryo extract, 10 parts of Sargassum polysaccharide, 13 parts of Cordyceps sinensis polysaccharide, 3 parts of Ganoderma lucidum polysaccharide, and 4 parts of Sargassum fusiformis polysaccharide.

[0078] Comparative Example 3

[0079] The difference from Example 2 is that the blood sugar lowering composition provided in Comparative Example 1 includes 20 parts of Matsutake polysaccharide, 10 parts of Schisandra chinensis extract, 10 parts of Sargassum polysaccharide, 8 parts of Cordyceps polysaccharide, 5 parts of Poria polysaccharide, and 3 parts of Ganoderma lucidum polysaccharide in parts by weight.

[0080] Comparative Example 4

[0081] The present comparative example provides a blood sugar lowering composition, which includes, by weight, 30 parts of matsutake polysaccharide, 10 parts of schisandra extract, 4 parts of lotus root extract, 1 part of lotus leaf extract, 3 parts of lotus embryo extract, 8 parts of cordyceps polysaccharide, 5 parts of tuckahoe polysaccharide, 3 parts of ganoderma lucidum polysaccharide, and 4 parts of sargassum polysaccharide.

[0082] Experimental Example 1

[0083] Method for establishing insulin-resistant (IR) HepG2 cells

[0084] HepG2 cells were cultured at 5×10 4 The cells were inoculated in a 96-well microplate at a density of 100 cells / well, and the sample volume was 100 μL DMEM high-glucose medium. After incubation for 12 h, the cells were washed with PBS and incubated at 100

[0085] μL / well of DMEM high-glucose medium containing 1.0 μM insulin was added to replace the original medium and cultured for 36 hours to obtain an insulin-resistant HepG2 cell model.

[0086] Insulin resistance (IR) HepG2 cells were plated into eight 6-well plates and the cell density was adjusted to 2.5×10 5 Each well contained 2 mL of DMEM high-glucose medium. HepG2 cells were plated in a 6-well plate and the cell density was adjusted to 2.5 × 10 5 / well, each well containing 2mL of DMEM culture medium. After incubation for 12 hours, 9 6-well plates were divided into 9 groups, namely normal control group, model control group, Example 2 to 4 groups, and comparative example 1 to 4 groups. The cells in each group were washed three times with PBS, and the cells in each group were treated with 2mL of the corresponding hypoglycemic composition solution with a final concentration of 2.0mg / mL per well. Among them, the model control group and the blank control group were not treated with the hypoglycemic composition. The cells in the model control group were cultured in DMEM high-glucose culture medium, and the blank control group was cultured in DMEM culture medium. After treatment, after incubation for 24 hours, the cells were collected and treated with cell lysis buffer for 10 minutes. The cell lysis buffer contained 20mM Tris buffer (pH7.5), 150mM NaCl and 1% TritonX-100 by mass. The glycogen content, hexokinase (HK) activity, and pyruvate kinase (PK) activity were detected by enzyme-linked immunosorbent assay (ELISA) kit according to the method in the kit instructions, and the results were measured. Figure 2 , Figure 3 , Figure 4 The supernatant of cells from each group of wells was collected, and the glucose concentration in the supernatant was determined using a glucose test kit according to the operating method in the kit instructions. The results are shown in Figure 1 shown.

[0087] Insulin resistance usually leads to decreased glucose utilization and downregulation of the expression of hepatic glycolytic enzymes, such as hexokinase (HK) and pyruvate kinase (PK). Hexokinase is the first key enzyme in glycolysis and the first rate-limiting enzyme in the formation of glycogen from glucose. Pyruvate kinase is another key enzyme in glycolysis that catalyzes the conversion of phosphoenolpyruvate (PEP) to pyruvate. In the diabetic state, the deficiency of these enzymes leads to the failure of glucose utilization.

[0088] like Figure 1-4 As shown in the figure, the glucose consumption level in the model control group was much lower than that in the normal control group, indicating that the model group was successfully established. Figure 1Compared with the model control group, the glucose consumption of Example 2 to 4 groups and Comparative Example 1 to 4 groups was significantly higher than that of the model control group, and the glucose consumption of Comparative Example 1 group was significantly lower than that of Example 2, indicating that in Example 2, Tricholoma matsutake polysaccharides and Schisandra chinensis extracts have a synergistic effect, synergistically promote the repair of pancreatic β cells, significantly improve insulin sensitivity, and promote the utilization of glucose by liver cells, thereby significantly improving glucose consumption; the glucose consumption of Comparative Example 2 group was equivalent to that of Example 2. Since it was an in vitro cell experiment, Cordyceps sinensis polysaccharides and Poria cocos polysaccharides did not affect glucose consumption; the blood sugar lowering effect of the comparative example 3 group is significantly lower than that of Example 2, indicating that in the comparative example 3, without adding lotus root extract, lotus leaf extract and lotus seed embryo extract, the blood sugar lowering effect is worse, indicating that fat metabolism disorder has a significant effect on the blood sugar lowering effect; the blood sugar lowering effect of the comparative example 4 group is lower than that of Example 2, indicating that in Example 2, Matsutake polysaccharide and Sargassum polysaccharide have a synergistic effect, synergistically improving the glucose consumption, glycogen synthesis and pyruvate kinase (PK) and hexokinase (HK) activities of insulin-resistant HepG2 cells, inhibiting insulin resistance and promoting the blood sugar lowering effect.

[0089] Figure 2 Compared with the model control group, the hexokinase activities of Example 2-4 groups and Comparative Examples 1-4 groups were significantly higher than those of the model control group, and the hexokinase activity of Comparative Example 1 group was significantly lower than that of Example 2, indicating that in Example 2, Tricholoma matsutake polysaccharides and Schisandra chinensis extracts have a synergistic effect, synergistically promote the repair of pancreatic β cells, significantly improve insulin sensitivity, and thus improve the activity of hexokinase; the activity of hexokinase in Comparative Example 2 group is equivalent to that in Example 2. Since it is an in vitro cell experiment, Cordyceps sinensis polysaccharides and Poria cocos polysaccharides do not affect the activity of hexokinase; Comparative Example 2 The blood sugar lowering effect of Example 3 is significantly lower than that of Example 2, indicating that in Example 3, without adding lotus root extract, lotus leaf extract, and lotus seed embryo extract, the blood sugar lowering effect is worse, indicating that fat metabolism disorder has a significant effect on the blood sugar lowering effect; the blood sugar lowering effect of Example 4 is lower than that of Example 2, indicating that in Example 2, Matsutake polysaccharides and Sargassum polysaccharides have a synergistic effect, synergistically improving the glucose consumption, glycogen synthesis, and pyruvate kinase (PK) and hexokinase (HK) activities of insulin-resistant HepG2 cells, inhibiting insulin resistance, and promoting the blood sugar lowering effect.

[0090] Figure 3Compared with the model control group, the pyruvate kinase activities of Example 2-4 groups and Comparative Examples 1-4 groups were significantly higher than those of the model control group, and the pyruvate kinase activity of Comparative Example 1 group was significantly lower than that of Example 2, indicating that in Example 2, Tricholoma matsutake polysaccharides and Schisandra chinensis extracts have a synergistic effect, synergistically promote the repair of pancreatic β cells, significantly improve insulin sensitivity, and thus improve pyruvate kinase activity; the pyruvate kinase activity of Comparative Example 2 group is equivalent to that of Example 2, and because it is an in vitro cell experiment, Cordyceps sinensis polysaccharides and Poria cocos polysaccharides do not affect the pyruvate kinase activity; The blood sugar lowering effect of Comparative Example 3 is significantly lower than that of Example 2, indicating that in Comparative Example 2, without adding lotus root extract, lotus leaf extract, and lotus seed embryo extract, the blood sugar lowering effect is worse, indicating that fat metabolism disorder has a significant effect on the blood sugar lowering effect; the blood sugar lowering effect of Comparative Example 4 is lower than that of Example 2, indicating that in Example 2, Matsutake polysaccharides and Sargassum polysaccharides have a synergistic effect, synergistically improving the glucose consumption, glycogen synthesis, and activity of pyruvate kinase (PK) and hexokinase (HK) in insulin-resistant HepG2 cells, inhibiting insulin resistance, and promoting the blood sugar lowering effect.

[0091] Figure 4 Compared with the model control group, the glycogen content of Example 2 to 4 groups and Comparative Example 1 to 4 groups were significantly higher than that of the model control group, and the glycogen content of Comparative Example 1 group was significantly lower than that of Example 2, indicating that in Example 2, Tricholoma matsutake polysaccharide and Schisandra chinensis extract had a synergistic effect, synergistically promoted the repair of pancreatic β cells, significantly improved insulin sensitivity, and thus increased the glycogen content; the glycogen content of Comparative Example 2 group was equivalent to that of Example 2. Since it was an in vitro cell experiment, Cordyceps sinensis polysaccharide and Poria cocos polysaccharide did not affect the glycogen content; Comparative Example 3 The hypoglycemic effect of the comparative example 4 group was significantly lower than that of Example 2, indicating that in comparative example 2, without adding lotus root extract, lotus leaf extract and lotus seed embryo extract, the hypoglycemic effect was worse, indicating that fat metabolism disorder has a significant effect on the hypoglycemic effect; the hypoglycemic effect of the comparative example 4 group was lower than that of Example 2, indicating that in Example 2, Matsutake polysaccharides and Sargassum polysaccharides have a synergistic effect, synergistically improving the glucose consumption, glycogen synthesis and pyruvate kinase (PK) and hexokinase (HK) activities of insulin-resistant HepG2 cells, inhibiting insulin resistance and promoting the hypoglycemic effect.

[0092] Experimental Example 2

[0093] 90 healthy mice, weighing 18 ± 2g, were purchased from the Experimental Animal Center of Guangdong Medical University. Ten normal control mice were used as the normal control group, and another 80 mice were used as the model group. Male and female mice were randomly selected. 55 mg of streptozotocin was dissolved in a 0.05M citric acid buffer solution with a pH of 4.5. Diabetes was induced by gavage. After 72 hours of gavage, the mice in the model control group that had fasted for 12 hours were measured for fasting blood sugar. Blood sugar values ​​greater than 10 mmol / L indicated that the diabetic model mice were successfully constructed. During the model construction process, the mice in the normal control group were injected with sterilized saline. The model group was divided into 8 groups, each with 10 mice, namely the model control group, Example 2-4 group, and Comparative Example 1-4 group. The blood sugar values ​​of each group were measured, and the average value was taken. The results are shown in Table 1.

[0094] Table 1 Blood glucose measurement results

[0095] Group Number of mice Blood glucose concentration (mmol / L) Normal control group 10 2.35±0.38 Model control group 10 18.56±3.25 Example 2 10 2.56±0.65 Example 3 10 2.32±0.69 Example 4 10 2.25±0.21 Comparative Example 1 10 5.31±1.52 Comparative Example 2 10 4.58±1.21 Comparative Example 3 10 13.56±1.02 Comparative Example 4 10 3.86±1.58

[0096] As can be seen from Table 1, the blood glucose value of the model control group is greater than 10mmol / L, indicating that the model was successfully constructed. Compared with the model control group, Examples 2 to 4 groups can all play a significant hypoglycemic effect, and Comparative Examples 1 to 3 groups can all play a significant hypoglycemic effect. The hypoglycemic effect of Comparative Example 1 group is lower than that of Example 2, indicating that in Example 1, Matsutake polysaccharides and Schisandra extract have a synergistic effect, synergistically promote the repair of pancreatic β cells, and enhance the therapeutic effect on type I diabetes; the hypoglycemic effect of Comparative Example 2 group is lower than that of Example 2, indicating that in Example 1, Cordyceps polysaccharides and Poria cocos polysaccharides have a synergistic effect, synergistically reduce the permeability of intestinal epithelial cells in diabetic mice, thereby promoting the reduction of type I diabetes. The effect on blood sugar; the blood sugar lowering effect of the comparative example 3 group is lower than that of Example 2, indicating that in Example 1, without adding lotus root extract, lotus leaf extract and lotus seed embryo extract, the blood sugar lowering effect is worse, indicating that fat metabolism disorder has an impact on the blood sugar lowering effect; the blood sugar lowering effect of the comparative example 4 group is lower than that of Example 2, indicating that in Example 1, Matsutake polysaccharides and Sargassum polysaccharides have a synergistic effect, synergistically improving the glucose consumption, glycogen synthesis and pyruvate kinase (PK) and hexokinase (HK) activities of insulin-resistant HepG2 cells, inhibiting insulin resistance and promoting the blood sugar lowering effect.

[0097] The above-mentioned embodiments are only preferred embodiments of the present invention and cannot be used to limit the scope of protection of the present invention. Any non-substantial changes and substitutions made by technicians in this field on the basis of the present invention shall fall within the scope of protection required by the present invention.

Claims

1. A blood sugar lowering composition, characterized in that: The invention is composed of the following raw materials in parts by weight: 15-20 parts of Matsutake polysaccharide, 10-15 parts of Schisandra chinensis extract, 3-5 parts of lotus root extract, 1-2 parts of lotus leaf extract, 1-3 parts of lotus seed embryo extract, 8-10 parts of Sargassum polysaccharide, 8-10 parts of Cordyceps polysaccharide, 5-8 parts of Poria polysaccharide, 2-5 parts of Ganoderma lucidum polysaccharide and 3-5 parts of Sargassum fusiformis polysaccharide.

2. A hypoglycemic composition according to claim 1, characterized in that: The invention is composed of the following raw materials in parts by weight: 20 parts of Matsutake polysaccharide, 10 parts of Schisandra chinensis extract, 4 parts of lotus root extract, 1 part of lotus leaf extract, 3 parts of lotus seed embryo extract, 10 parts of Sargassum polysaccharide, 8 parts of Cordyceps sinensis polysaccharide, 5 parts of Poria polysaccharide, 3 parts of Ganoderma lucidum polysaccharide and 4 parts of Sargassum fusiforme polysaccharide.

3. A method for preparing a hypoglycemic composition according to any one of claims 1 to 2, characterized in that: The following steps are involved: The Tricholoma matsutake polysaccharide, Schisandra chinensis extract, lotus root extract, lotus leaf extract, lotus seed embryo extract, Sargassum polysaccharide, Cordyceps sinensis polysaccharide, Poria polysaccharide, Ganoderma lucidum polysaccharide and Sargassum fusiformis polysaccharide are weighed according to the formula amount and dissolved in 70% (V / V) ethanol aqueous solution in sequence, and freeze-dried to obtain the blood sugar lowering composition.

4. Use of a hypoglycemic composition according to any one of claims 1 to 2 in the preparation of a hypoglycemic drug, characterized in that: The hypoglycemic composition is used as an active ingredient and a pharmaceutically acceptable drug excipient to prepare the hypoglycemic drug; the drug excipient is one of an excipient, a disintegrant, a sweetener, a binder, a coating agent and a bulking agent.

5. Use of a hypoglycemic composition according to any one of claims 4 in the preparation of a hypoglycemic drug, characterized in that: The blood sugar lowering drug is one of a solution, an emulsion, a suspension, an injection, a powder, a granule, a tablet, and a pill.

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