Traditional Chinese medicine composition, preparation method thereof and application of traditional Chinese medicine composition in preparation of hypoglycemic products

Abstract

The invention provides a traditional Chinese medicine composition, a preparation method thereof and application of the traditional Chinese medicine composition in preparation of hypoglycemic products, and relates to the technical field of traditional Chinese medicines. The invention discloses a traditional Chinese medicine composition. The traditional Chinese medicine composition is prepared from the following raw materials: 4-8 parts of ginseng stem and leaf saponin, 8-16 parts of astragalus polysaccharide, 6-12 parts of astragaloside, 8-16 parts of catalpol, 6-12 parts of rehmannia D, 8-16 parts of verbascoside, 3-5 parts of raspberry polysaccharide, 4-8 parts of ellagic acid, 4-8 parts of pachymaran, 5-10 parts of pachymic acid, 8-16 parts of lycium barbarum polysaccharide, 40-62 parts of schisandra chinensis, 20-40 parts of Chinese yam, 40-62 parts of radix ophiopogonis, 40-62 parts of radix trichosanthis and 40-62 parts of rhizoma alismatis. The traditional Chinese medicine raw materials are pretreated by using the high-voltage pulsed electric field, and then ethanol extraction treatment is performed twice, so that the yield of the traditional Chinese medicine extract is high, and the traditional Chinese medicine composition obtained by mixing the traditional Chinese medicine extract with other active ingredients has an excellent effect of treating type II diabetes mellitus.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, specifically relating to a traditional Chinese medicine composition and its preparation method, and its application in the preparation of hypoglycemic products. Background Technology

[0002] Diabetes is a common metabolic disease, and its incidence is related to genetic factors, lifestyle, diet, and age. Based on the pathological characteristics of patients, diabetes can be divided into two types: type 1 diabetes and type 2 diabetes, also known as non-insulin-dependent diabetes mellitus. The main characteristics of type 2 diabetes are decreased insulin secretion and reduced insulin action from pancreatic β-cells, i.e., insulin resistance.

[0003] Currently, most medications used clinically to treat type 2 diabetes, besides insulin, are chemical drugs, such as sulfonylureas, biguanides, other hypoglycemic agents, and adjuvant medications. Sulfonylureas are more likely to cause adverse reactions such as hypoglycemia, granulocytopenia, and cardiovascular disease; high doses of biguanides can cause gastrointestinal reactions and easily lead to lactic acidosis in patients with lung, liver, or kidney disease, posing a threat to the safety of medication use for many patients. Traditional Chinese medicine generally agrees on the etiology of diabetes, believing it is mainly due to factors such as excessive consumption of rich and sweet foods, excessive emotional stress, immoderate sexual activity, febrile diseases causing dryness and heat, and congenital deficiencies.

[0004] Currently, commonly used traditional Chinese medicines for lowering blood sugar include: Shenqi Jiangtang Granules, Jiangtangning Capsules, and Xiaoke Pills, all of which can be used for type II diabetes. Shenqi Jiangtang Granules are formulated with: ginseng (stem and leaf) saponins, schisandra, astragalus, yam, rehmannia, raspberry, ophiopogon, poria, trichosanthes root, alisma, and wolfberry. Its functions and indications are: tonifying qi and nourishing yin, invigorating the spleen and kidneys. Chinese invention patent CN10245198A describes the above formula, which is prepared into an effective preparation after different physical or chemical treatments based on the different physicochemical properties of the active ingredients of each herb. Chinese invention patent CN103223104A discloses a traditional Chinese medicine composition for treating diabetes and its preparation method, improving the original preparation method of Shenqi Jiangtang Granules, especially the extraction and refining methods of the four herbs trichosanthes root, yam, raspberry, and poria. It uses an ultra-fine pulverization method to improve drug absorption and bioavailability. However, in the existing technology, the extraction of other components is carried out by conventional alcohol extraction or water decoction, resulting in limited content of active ingredients in the extracted extracts and low utilization of effective active ingredients.

[0005] Furthermore, Chinese invention patent CN106266543A improves the composition of the original Astragalus and Ginseng Hypoglycemic Granules by adding Rehmannia glutinosa, Scrophularia ningpoensis, and Anemarrhena asphodeloides, providing a new formulation of a traditional Chinese medicine composition for treating type II diabetes. Chinese invention patent CN114848602A adds 10wt% extracts of Pueraria lobata, Ophiopogon japonicus, Angelica sinensis, and Crataegus pinnatifida to the original Astragalus compound hypoglycemic tablets formulation. These prior art improvements to traditional hypoglycemic formulations all involve altering the original formula and adding medicinal ingredients. The addition of these components may affect the principal, assistant, adjuvant, and guide principles of traditional Chinese medicine formulas, or weaken the efficacy of the original medicinal ingredients, or even cause antagonism, reducing the theoretical efficacy.

[0006] Therefore, further research is needed on the original Astragalus compound hypoglycemic tablet formulation and preparation method, and improvements should be made in terms of Chinese herbal extraction and active ingredients in order to significantly enhance the hypoglycemic efficacy of the original Chinese herbal medicine. Summary of the Invention

[0007] This invention addresses the problems existing in the prior art by providing a traditional Chinese medicine composition and its preparation method, as well as its application in the preparation of hypoglycemic products.

[0008] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0009] First, the present invention provides a traditional Chinese medicine composition, which, by weight, is made from the following raw materials: 4-8 parts ginseng stem and leaf saponins, 8-16 parts astragalus polysaccharide, 6-12 parts astragaloside A, 8-16 parts catalpol, 6-12 parts rehmannia glycoside D, 8-16 parts verbascoside, 3-5 parts raspberry polysaccharide, 4-8 parts ellagic acid, 4-8 parts poria polysaccharide, 5-10 parts poria acid, 8-16 parts wolfberry polysaccharide, 40-62 parts schisandra, 20-40 parts yam, 40-62 parts ophiopogon japonicus, 40-62 parts trichosanthes kirilowii and 40-62 parts alisma plantago-aquatica.

[0010] Preferably, the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 5-7 parts ginseng stem and leaf saponins, 10-14 parts astragalus polysaccharide, 8-11 parts astragaloside A, 10-14 parts catalpol, 7-10 parts rehmannia glycoside D, 10-14 parts verbascoside, 3.5-4.5 parts raspberry polysaccharide, 5-7 parts ellagic acid, 5-7 parts poria polysaccharide, 6-9 parts poria acid, 10-14 parts wolfberry polysaccharide, 50-60 parts schisandra, 25-35 parts yam, 45-55 parts ophiopogon japonicus, 45-55 parts trichosanthes kirilowii, and 45-55 parts alisma plantago-aquatica.

[0011] More preferably, the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 6 parts ginseng stem and leaf saponins, 12 parts astragalus polysaccharide, 10 parts astragaloside A, 12 parts catalpol, 8 parts rehmannia glycoside D, 12 parts verbascoside, 4 parts raspberry polysaccharide, 6 parts ellagic acid, 6 parts poria polysaccharide, 8 parts poria acid, 12 parts wolfberry polysaccharide, 55 parts schisandra, 30 parts yam, 50 parts ophiopogon japonicus, 50 parts trichosanthes kirilowii and 50 parts alisma plantago-aquatica.

[0012] Then, the present invention provides a method for preparing the above-mentioned traditional Chinese medicine composition, comprising the following steps: (1) Schisandra chinensis, Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica are pulverized, dried, and then pretreated with a high-voltage pulse electric field to obtain a pretreated mixture; The pretreated mixture is filtered to obtain filtrate A and filter residue A. (2) Filter residue A was initially extracted with ethanol with a volume concentration of 20%-40%, filtered, and filtrate B and filter residue B were obtained. (3) Filter residue B is extracted a second time with ethanol of 60%-80% by volume, filtered, and filtrate C and filter residue C are obtained. (4) Filtrate A, Filtrate B and Filtrate C are dried and mixed to obtain a Chinese medicine extract; then it is mixed with ginseng stem and leaf saponins, astragalus polysaccharide, astragaloside A, catalpol, rehmannia glycoside D, verbascoside, raspberry polysaccharide, ellagic acid, poria polysaccharide, poria acid and wolfberry polysaccharide to obtain a Chinese medicine composition.

[0013] Preferably, in step (1), the pulverization is performed to pulverize the material into coarse powder with a particle size of 150-500 μm.

[0014] Preferably, in step (1), the drying process involves a drying temperature ≤60℃ and a drying time ≤48h.

[0015] More preferably, in step (1), the drying temperature is 45±5℃ and the drying time is 12-36h; even more preferably, in step (1), the drying time is 24h.

[0016] Preferably, in step (1), the conditions for pretreatment of the high-voltage pulse electric field are: electric field strength 20-28kV / cm, pulse number 5-7, material-liquid ratio 1g:15-25mL, and pH value 7-9.

[0017] More preferably, in step (1), the conditions for pretreatment by the high-voltage pulse electric field are: electric field strength 24kV / cm, number of pulses 6, material-liquid ratio 1g:20mL, and pH value 8.

[0018] More preferably, in step (1), the pH value is adjusted using a conventional alkaline reagent, not limited to sodium carbonate, sodium hydroxide, or sodium bicarbonate.

[0019] Preferably, in step (2), the volume concentration of ethanol is 30%.

[0020] Preferably, in step (2), the initial extraction conditions are: extraction temperature 65-75℃, extraction time 2-4h, extraction material-liquid ratio 1g:25-35mL, and the weight of the material in the material-liquid ratio is the total weight of Schisandra chinensis, Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica in step (1).

[0021] More preferably, in step (2), the initial extraction conditions are: extraction temperature 70℃, extraction time 3h, and extraction material-to-liquid ratio 1g:30mL.

[0022] Preferably, in step (3), the volume concentration of the ethanol is 70%.

[0023] Preferably, in step (3), the conditions for the secondary extraction are: extraction temperature 50-60℃, extraction time 1-3h, extraction material-liquid ratio 1g:25-35mL, and the weight of the material in the material-liquid ratio is the total weight of Schisandra chinensis, Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica in step (1).

[0024] More preferably, in step (3), the conditions for the secondary extraction are: extraction temperature 55℃, extraction time 2h, and extraction material-to-liquid ratio 1g:30mL.

[0025] Preferably, in step (4), the drying temperature is ≤60℃ and the drying time is ≤48h.

[0026] More preferably, in step (4), the drying is freeze drying, and the drying time is 24 hours.

[0027] Preferably, in step (4), the liquid is concentrated under reduced pressure before drying until the density is 1.05-1.30 (60°C) to remove most of the liquid.

[0028] Finally, this invention provides the application of the above-mentioned traditional Chinese medicine composition in the preparation of hypoglycemic products.

[0029] Preferably, the product used in the application is a drug.

[0030] More preferably, the drug is a drug with hypoglycemic effect.

[0031] More preferably, the drug is a drug for treating type II diabetes.

[0032] More preferably, the drug has the effects of lowering blood sugar, treating diabetes, increasing glucose consumption of insulin-resistant Hepg2 cells, promoting the growth of insulin-resistant Hepg2 cells, lowering blood sugar levels, and increasing serum insulin levels.

[0033] Compared with the prior art, the present invention has the following beneficial effects: 1. This invention combines specific active ingredients and traditional Chinese medicine components to obtain a traditional Chinese medicine composition that can significantly improve glucose consumption in insulin-resistant Hepg2 cells, promote the growth of insulin-resistant Hepg2 cells, reduce blood glucose levels, and increase serum insulin levels, thus bringing excellent therapeutic effects on type II diabetes.

[0034] 2. In the traditional Chinese medicine composition of the present invention, the traditional Chinese medicine components Schisandra chinensis, Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica are pretreated with a high-voltage pulsed electric field and extracted with a gradient increase in ethanol concentration, which significantly improves the yield of the traditional Chinese medicine extract and increases the activity of the effective components, thereby bringing excellent hypoglycemic and diabetic symptoms improvement effects. Detailed Implementation

[0035] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0036] When numerical ranges are given in the embodiments, it should be understood that, unless otherwise stated in the invention, both endpoints of each numerical range and any value between the two endpoints may be selected. Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0037] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all chemical reagents used in the embodiments of the present invention were obtained through conventional commercial channels. Purchase information for the main components is shown in Table 1. Products from different manufacturers do not have a significant impact on the efficacy.

[0038] Table 1

[0039] Example 1 A traditional Chinese medicine composition, by weight, is made from the following raw materials: 6 parts ginseng stem and leaf saponins, 12 parts astragalus polysaccharide, 10 parts astragaloside A, 12 parts catalpol, 8 parts rehmannia glycoside D, 12 parts verbascoside, 4 parts raspberry polysaccharide, 6 parts ellagic acid, 6 parts poria polysaccharide, 8 parts poria acid, 12 parts wolfberry polysaccharide, 55 parts schisandra, 30 parts yam, 50 parts ophiopogon japonicus, 50 parts trichosanthes kirilowii and 50 parts alisma plantago-aquatica.

[0040] The preparation method of this traditional Chinese medicine composition is as follows: (1) Schisandra chinensis, Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica were pulverized into coarse powder with a particle size between 150-500 μm and dried at 45±5℃ for 24 h; then pretreated with a high voltage pulse electric field: electric field strength 24 kV / cm, pulse number 6, material-liquid ratio 1 g: 20 mL, pH value 8 (sodium carbonate was used to adjust the pH) to obtain a pretreated mixture; The pretreated mixture is filtered to obtain filtrate A and filter residue A. (2) Filter residue A was initially extracted with ethanol with a volume concentration of 30%, the extraction temperature was 70℃, the extraction time was 3h, and the extraction material-liquid ratio was 1g:30mL; filtration was performed to obtain filtrate B and filter residue B. The weight of the materials mentioned in the material-liquid ratio is the total weight of Schisandra chinensis, Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica in step (1); (3) Filter residue B was extracted a second time with 70% ethanol at a volume concentration of 55℃ for 2 hours and the ratio of extract to liquid was 1g:30mL. After filtration, filtrate C and filter residue C were obtained. The weight of the materials mentioned in the material-liquid ratio is the total weight of Schisandra chinensis, Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica in step (1); (4) Filtrate A, Filtrate B and Filtrate C were concentrated under reduced pressure to a density of 1.20 (60℃), and then freeze-dried for 24 hours to obtain the Chinese herbal extract; The Chinese herbal extracts and the prescribed amounts of ginseng stem and leaf saponins, astragalus polysaccharides, astragaloside A, catalpol, rehmannia glycoside D, verbascoside, raspberry polysaccharides, ellagic acid, poria polysaccharides, poria acid, and wolfberry polysaccharides are mixed to obtain a Chinese herbal composition.

[0041] Example 2 A traditional Chinese medicine composition, by weight, is made from the following raw materials: 5 parts ginseng stem and leaf saponins, 10 parts astragalus polysaccharide, 11 parts astragaloside A, 10 parts catalpol, 10 parts rehmannia glycoside D, 14 parts verbascoside, 3.5 parts raspberry polysaccharide, 7 parts ellagic acid, 7 parts poria polysaccharide, 6 parts poria acid, 14 parts wolfberry polysaccharide, 60 parts schisandra, 35 parts yam, 45 parts ophiopogon japonicus, 45 parts trichosanthes kirilowii and 45 parts alisma plantago-aquatica.

[0042] The preparation method of this traditional Chinese medicine composition is as follows: (1) Schisandra chinensis, Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica were pulverized into coarse powder with a particle size between 150-500 μm and dried at 45±5℃ for 24 h; then pretreated with a high voltage pulse electric field: electric field strength 28 kV / cm, pulse number 7, material-liquid ratio 1 g:15 mL, pH value 7 (sodium carbonate was used to adjust the pH) to obtain a pretreated mixture; The pretreated mixture is filtered to obtain filtrate A and filter residue A. (2) Filter residue A was initially extracted with 40% ethanol at a volume concentration of 65℃ for 2 hours at a material-to-liquid ratio of 1g:35mL; filtration was performed to obtain filtrate B and filter residue B. The weight of the materials mentioned in the material-liquid ratio is the total weight of Schisandra chinensis, Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica in step (1); (3) Filter residue B was extracted a second time with 60% ethanol at a volume concentration of 60℃ for 3 hours and the ratio of extract to liquid was 1g:25mL. After filtration, filtrate C and filter residue C were obtained. The weight of the materials mentioned in the material-liquid ratio is the total weight of Schisandra chinensis, Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica in step (1); (4) Filtrate A, Filtrate B and Filtrate C were concentrated under reduced pressure to a density of 1.05 (60℃), and then freeze-dried for 24 hours to obtain the Chinese herbal extract; The Chinese herbal extracts and the prescribed amounts of ginseng stem and leaf saponins, astragalus polysaccharides, astragaloside A, catalpol, rehmannia glycoside D, verbascoside, raspberry polysaccharides, ellagic acid, poria polysaccharides, poria acid, and wolfberry polysaccharides are mixed to obtain a Chinese herbal composition.

[0043] Example 3 A traditional Chinese medicine composition, by weight, is made from the following raw materials: 7 parts ginseng stem and leaf saponins, 14 parts astragalus polysaccharide, 8 parts astragaloside A, 14 parts catalpol, 7 parts rehmannia glycoside D, 10 parts verbascoside, 4.5 parts raspberry polysaccharide, 5 parts ellagic acid, 5 parts poria polysaccharide, 9 parts poria acid, 10 parts wolfberry polysaccharide, 50 parts schisandra, 25 parts yam, 55 parts ophiopogon japonicus, 55 parts trichosanthes kirilowii and 55 parts alisma plantago-aquatica.

[0044] The preparation method of this traditional Chinese medicine composition is as follows: (1) Schisandra chinensis, Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica were pulverized into coarse powder with a particle size between 150-500 μm and dried at 45±5℃ for 24 h; then pretreated with a high-voltage pulse electric field: electric field strength 20 kV / cm, pulse number 5, material-liquid ratio 1 g: 25 mL, pH value 9 (sodium carbonate was used to adjust the pH) to obtain a pretreated mixture; The pretreated mixture is filtered to obtain filtrate A and filter residue A. (2) Filter residue A was initially extracted with 20% ethanol at a volume concentration of 75°C for 4 hours at a material-to-liquid ratio of 1g:25mL; then filtered to obtain filtrate B and filter residue B. The weight of the materials mentioned in the material-liquid ratio is the total weight of Schisandra chinensis, Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica in step (1); (3) Filter residue B was extracted a second time with 80% ethanol at a volume concentration of 50°C for 1 hour and the ratio of extract to liquid was 1 g: 35 mL. The residue was then filtered to obtain filtrate C and filter residue C. The weight of the materials mentioned in the material-liquid ratio is the total weight of Schisandra chinensis, Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica in step (1); (4) Filtrate A, Filtrate B and Filtrate C were concentrated under reduced pressure to a density of 1.30 (60℃), and then freeze-dried for 24 hours to obtain the Chinese herbal extract; The Chinese herbal extracts and the prescribed amounts of ginseng stem and leaf saponins, astragalus polysaccharides, astragaloside A, catalpol, rehmannia glycoside D, verbascoside, raspberry polysaccharides, ellagic acid, poria polysaccharides, poria acid, and wolfberry polysaccharides are mixed to obtain a Chinese herbal composition.

[0045] Example 4 Unlike Example 1, the weight parts of the traditional Chinese medicine composition are different.

[0046] A traditional Chinese medicine composition, by weight, is made from the following raw materials: 4 parts ginseng stem and leaf saponins, 8 parts astragalus polysaccharide, 12 parts astragaloside A, 8 parts catalpol, 12 parts rehmannia glycoside D, 16 parts verbascoside, 3 parts raspberry polysaccharide, 8 parts ellagic acid, 8 parts poria polysaccharide, 5 parts poria acid, 16 parts wolfberry polysaccharide, 62 parts schisandra, 40 parts yam, 40 parts ophiopogon japonicus, 40 parts trichosanthes kirilowii and 40 parts alisma plantago-aquatica.

[0047] Everything else is the same as in Example 1.

[0048] Example 5 Unlike Example 1, the weight parts of the traditional Chinese medicine composition are different.

[0049] A traditional Chinese medicine composition, by weight, is made from the following raw materials: 8 parts ginseng stem and leaf saponins, 16 parts astragalus polysaccharide, 6 parts astragaloside A, 16 parts catalpol, 6 parts rehmannia glycoside D, 8 parts verbascoside, 5 parts raspberry polysaccharide, 4 parts ellagic acid, 4 parts poria polysaccharide, 10 parts poria acid, 8 parts wolfberry polysaccharide, 40 parts schisandra, 20 parts yam, 62 parts ophiopogon japonicus, 62 parts trichosanthes kirilowii and 62 parts alisma plantago-aquatica.

[0050] Everything else is the same as in Example 1.

[0051] Comparative Example 1 The difference from Example 1 is that verbascoside is replaced with aucubin, otherwise it is the same as Example 1.

[0052] A traditional Chinese medicine composition, by weight, is made from the following raw materials: 6 parts ginseng stem and leaf saponins, 12 parts astragalus polysaccharide, 10 parts astragaloside A, 12 parts catalpol, 8 parts rehmannia glycoside D, 12 parts aucubin, 4 parts raspberry polysaccharide, 6 parts ellagic acid, 6 parts poria polysaccharide, 8 parts poria acid, 12 parts wolfberry polysaccharide, 55 parts schisandra, 30 parts yam, 50 parts ophiopogon japonicus, 50 parts trichosanthes kirilowii and 50 parts alisma plantago-aquatica.

[0053] Comparative Example 2 The difference from Example 1 is that pachymic acid is replaced with ellagic acid, while the rest is the same as in Example 1.

[0054] A traditional Chinese medicine composition, by weight, is made from the following raw materials: 6 parts ginseng stem and leaf saponins, 12 parts astragalus polysaccharide, 10 parts astragaloside A, 12 parts catalpol, 8 parts rehmannia glycoside D, 12 parts verbascoside, 4 parts raspberry polysaccharide, 14 parts ellagic acid, 6 parts poria polysaccharide, 12 parts wolfberry polysaccharide, 55 parts schisandra, 30 parts yam, 50 parts ophiopogon japonicus, 50 parts trichosanthes kirilowii and 50 parts alisma plantago-aquatica.

[0055] Comparative Example 3 The difference from Example 1 is that raspberry polysaccharide is replaced with wolfberry polysaccharide, otherwise it is the same as Example 1.

[0056] A traditional Chinese medicine composition, by weight, is made from the following raw materials: 6 parts ginseng stem and leaf saponins, 12 parts astragalus polysaccharide, 10 parts astragaloside A, 12 parts catalpol, 8 parts rehmannia glycoside D, 12 parts verbascoside, 6 parts ellagic acid, 6 parts poria polysaccharide, 8 parts poria acid, 16 parts wolfberry polysaccharide, 55 parts schisandra, 30 parts yam, 50 parts ophiopogon japonicus, 50 parts trichosanthes kirilowii and 50 parts alisma plantago-aquatica.

[0057] Comparative Example 4 Unlike Example 1, the raw materials in the traditional Chinese medicine composition are different by weight.

[0058] A traditional Chinese medicine composition, by weight, is made from the following raw materials: 6 parts ginseng stem and leaf saponins, 2 parts astragalus polysaccharide, 13 parts astragaloside A, 4 parts catalpol, 6 parts rehmannia glycoside D, 4 parts verbascoside, 2 parts raspberry polysaccharide, 1 part ellagic acid, 6 parts poria polysaccharide, 8 parts poria acid, 44 parts wolfberry polysaccharide, 31 parts schisandra, 111 parts yam, 31 parts ophiopogon japonicus, 31 parts trichosanthes kirilowii and 31 parts alisma plantago-aquatica.

[0059] Everything else is the same as in Example 1.

[0060] Comparative Example 5 Unlike Example 1, the preparation method of the traditional Chinese medicine composition is different. The pretreatment process using a high-voltage pulsed electric field in step (1) is replaced with microwave-assisted treatment.

[0061] The preparation method of this traditional Chinese medicine composition is as follows: (1) Schisandra chinensis, Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica were pulverized into coarse powder with a particle size between 150-500 μm and dried at 45±5℃ for 24 h; then microwave-assisted pretreatment was carried out: microwave power 100W, material-liquid ratio 1g:20mL, pH value 8 (sodium carbonate was used to adjust the pH) for 20 min to obtain a pretreated mixture; The pretreated mixture is filtered to obtain filtrate A and filter residue A. Steps (2)-(4) are the same as in Example 1.

[0062] Comparative Example 6 Unlike Example 1, the preparation method of the traditional Chinese medicine composition is different. The extraction of the traditional Chinese medicine was carried out using the water decoction and ethanol extraction method described in publication number CN103223104A.

[0063] The preparation method of this traditional Chinese medicine composition is as follows: (1) The Schisandra chinensis was crushed into coarse powder with a particle size between 150-500 μm and dried at 45±5℃ for 24h; it was then extracted twice by reflux with 50% ethanol for 2 hours each time, and the ethanol extract was concentrated under reduced pressure to obtain a Schisandra chinensis ethanol extract with a relative density of 1.30 (60℃). (2) Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica are pulverized into coarse powder with a particle size between 150-500μm and dried at 45±5℃ for 24h; water is used for initial extraction, and the extraction is carried out twice by boiling, each time for 2h, with an extraction material-to-liquid ratio of 1g:30mL; the mixture is filtered to obtain filtrate B and residue B. The weight of the materials mentioned in the material-liquid ratio is the total weight of yam, ophiopogon japonicus, alisma plantago-aquatica, and trichosanthes kirilowii. Filtrate B was concentrated under reduced pressure to obtain an extract with a relative density of 1.30 (60°C); (3) The extract from step (2) is added to ethanol until the volume concentration reaches 65% for resolution and purification, and concentrated under reduced pressure to a water extract with a relative density of 1.30 (60℃). (4) Mix the schisandra alcohol extract from step (1) and the water extract from step (3), freeze-dry for 24 hours to obtain the Chinese herbal extract; The Chinese herbal extracts and the prescribed amounts of ginseng stem and leaf saponins, astragalus polysaccharides, astragaloside A, catalpol, rehmannia glycoside D, verbascoside, raspberry polysaccharides, ellagic acid, poria polysaccharides, poria acid, and wolfberry polysaccharides are mixed to obtain a Chinese herbal composition.

[0064] Comparative Example 7 Unlike Example 1, the ethanol in steps (2) and (3) was replaced with ethanol with a volume concentration of 50%.

[0065] Everything else is the same as in Example 1.

[0066] Experiment 1 Yield of Traditional Chinese Medicine Extracts Yield of Chinese herbal extract (%) = mass of Chinese herbal extract / total mass of Chinese herbal raw materials (Schisandra chinensis, Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica) × 100%.

[0067] The yields of the herbal extracts prepared in Examples 1-3 and Comparative Examples 5-7 were calculated. The experiments were conducted in parallel for three times, and the average yield results are shown in Table 2.

[0068] Table 2

[0069] In Table 2, # This indicates that each group has a significant difference compared to Group 1 of Example 1. # This indicates that P < 0.05.

[0070] As can be seen from Table 2, the herbal extract prepared by the method of the present invention has a higher yield compared to other conventional extraction methods, with the yield improvement rate reaching half or more of the comparative example. Therefore, the preparation method of the present invention brings about a significant improvement in yield performance.

[0071] Experiment 2: Effects of Traditional Chinese Medicine Composition on Glucose Consumption and Cell Proliferation in Insulin-Resistant Hepg2 Cells 1. Experimental Objective The effects of traditional Chinese medicine compositions on an insulin-resistant Hepg2 cell model were determined to provide a basis for selecting effective hypoglycemic drugs.

[0072] 2. Reagents and instruments used in the experiment DMEM medium; fetal bovine serum; trypsin; PBS; glucose kit; bovine insulin; MTT; cell culture incubator; ELISA reader; microscope.

[0073] 3. Experimental cells Human hepatocellular carcinoma Hepg2 cell line.

[0074] 4. Preparation of sample solution (1) Preparation of insulin (M=5733.52) stock solution: Weigh 1.15 mg insulin into a 5 mL test tube, add 1 mL PBS, dissolve in a warm water bath, filter through a 0.22 μm microporous membrane to obtain a concentration of 2 × 10⁻⁶. -4 1 mol / L stock solution, dispensed for later use.

[0075] (2) Preparation of metformin (M=165.63) stock solution: Weigh 662.52 mg of metformin raw material into a 5 mL test tube, add 1 mL of PBS, dissolve in warm water, filter through a 0.22 μm microporous membrane, and dispense for later use.

[0076] (3) Preparation of stock solution of traditional Chinese medicine composition: Take 1 mg of traditional Chinese medicine composition, add 100 μL of DMSO to dissolve it, and prepare a stock solution of traditional Chinese medicine composition with a final concentration of 10 g / L for later use.

[0077] 5. Experimental Methods (1) Hepg2 cell culture After resuscitation, Hepg2 cells were cultured in high-glucose DMEM medium containing 10% fetal bovine serum at 37°C and 5% CO2. Once the cells reached confluence, the culture medium was discarded, and the cells were gently washed twice with PBS. The cells were then digested with trypsin and EDTA, and passaged every 3 days at a 1:3 ratio. Cells in the logarithmic growth phase were used for experiments.

[0078] (2) Effects of traditional Chinese medicine extracts on glucose consumption in insulin-resistant Hepg2 cells After digesting the cells in the logarithmic growth phase, the cell concentration was adjusted to 5 × 10⁶ cells / year using high-glucose DMEM medium containing 10% fetal bovine serum. 4 Cells were seeded at a density of 200 μL / mL in 96-well plates. Each vertical row constituted a group, including a blank control group, a model group, a metformin positive control group (final concentration 2 mmol / L), and a traditional Chinese medicine composition group (Example 1-Example 5, Comparative Example 1-Comparative Example 7; each group's dose concentration was 1×10⁻⁶). -6 After culturing for 24 hours, the culture medium was discarded, and the insulin was washed twice with PBS. The insulin was then prepared into a concentration of 1×10⁻⁶ g / L using serum-free, high-glucose DMEM medium. -7 1 mmol / L drug diluent. For the traditional Chinese medicine (TCM) composition groups, each TCM composition was diluted to the corresponding detection dose concentration using this drug diluent, and the final DMSO concentration in each experimental group was controlled at 0.01%, 100 mL per well, and incubated for 24 h. For the metformin positive control group, metformin was diluted to the corresponding detection dose concentration (final concentration 2 mmol / L) using the drug diluent, and the final DMSO concentration in each experimental group was controlled at 0.01%, 100 mL per well, and incubated for 24 h. The model group was cultured with 1 μL / mL DMSO for 24 h. The blank control group consisted of normally cultured cells.

[0079] After 24 hours, the culture medium was centrifuged at 1000 rpm for 5 minutes, and the supernatant was used to determine the residual glucose content. A blank control group was used as a control, and the glucose consumption of cells in each group was calculated. Simultaneously, the effect of the traditional Chinese medicine composition on cell growth activity was detected using the MTT assay.

[0080] (3) Effect of MTT assay on cell growth activity of traditional Chinese medicine composition After the glucose consumption experiment, the test culture medium was removed, and 80 μL of DMEM cell culture medium and 20 μL of LTT were added to each well. The cells were then incubated at 37°C with 5% CO2. After 4 hours, the culture was terminated, the liquid in the wells was carefully discarded, and 100 μL of DMSO was added to each well. The cells were shaken at room temperature for 1 minute to fully dissolve the crystals. The absorbance of each well was measured at 492 nm using a microplate reader to detect the effect on cell growth activity.

[0081] The effects of the traditional Chinese medicine composition on glucose consumption and cell growth activity in insulin-resistant Hepg2 cells are shown in Table 3 (x±s, n=4).

[0082] Table 3

[0083] In Table 3, # and ## This indicates that the data in the same column shows significant differences between each group and the model group. # This indicates that P < 0.05. ## This indicates that P < 0.01; △ and △△ This indicates that the data in the same column shows significant differences compared to the group in Example 1. △ This indicates that P < 0.05. △△ This indicates that P < 0.01.

[0084] As shown in Table 3, the herbal extract of this invention has a significant hypoglycemic effect, significantly increasing glucose consumption. Compared with comparative examples 1-3 (components with replaced components), comparative example 4 (component ratios changed), and comparative examples 5-7 (preparation methods changed), glucose consumption is significantly increased, demonstrating excellent hypoglycemic effect. The herbal extract of this invention also promotes cell growth, with no observed cytotoxic effects.

[0085] Experiment 3: Effects of Traditional Chinese Medicine Composition on Insulin-Resistant Rats 1. Experimental Materials 1.1 Animals Wistar rats, male, SPF grade, 130 rats, 6-8 weeks old, weighing 200±20g, were purchased from Spiford (Beijing) Biotechnology Co., Ltd.

[0086] Breeding environment: room temperature 20-26℃, relative humidity controlled at 40-70%, animal stocking density is 2 animals / cage.

[0087] 1.2 Drugs The traditional Chinese medicine compositions prepared in Examples 1-5, and the traditional Chinese medicine compositions prepared in Comparative Examples 1-4 and Comparative Examples 6; metformin hydrochloride tablets, streptozotocin (STZ), sodium citrate, citric acid; basic feed, high-sugar and high-fat feed.

[0088] 1.3 Instruments Blood glucose meter, insulin test kit.

[0089] 2. Dosage design The oral dose of the traditional Chinese medicine composition is 80 mg / kg; the oral dose of metformin hydrochloride is 104 mg / kg; once daily.

[0090] 3. Experimental Methods (1) Animal modeling of type II diabetes mellitus (T2DM): Wistar rats were acclimatized for 7 days with free access to food and water, and were randomly divided into a modeling group (n=110) and a normal group (n=10) according to their body weight. The normal group was fed a basal diet, while the modeling group was fed a high-sugar, high-fat diet. Both groups had free access to food and water for 8 weeks. After 8 weeks, the rats were fasted but allowed free access to water for 12 hours, and venous blood was collected to measure blood glucose and insulin levels. The modeling group received a single intraperitoneal injection of streptozotocin (STZ) 25 mg / kg, which was dissolved in citrate buffer (pH 4.2-4.5) at a concentration of 2% before injection. The normal group received an intraperitoneal injection of the same volume of citrate-sodium citrate buffer. Fasting blood glucose and serum insulin levels were measured 72 hours later. A fasting blood glucose level >16.7 mmol / L was considered a successful model.

[0091] (2) Grouping and administration: Rats that successfully developed the model were randomly divided into a model group, a positive drug group, and a traditional Chinese medicine composition group (Example 1-Example 5, Comparative Example 1-Comparative Example 4, and Comparative Example 6) according to their blood glucose levels, with 10 rats in each group. The normal group was given a basal diet, while the other groups continued to be given a high-sugar, high-fat diet. The normal group and the model group were given purified water by gavage; the positive drug group was given metformin hydrochloride by gavage; and the traditional Chinese medicine composition groups were given the corresponding doses of the traditional Chinese medicine composition.

[0092] (3) Measurement of relevant indicators: After 8 weeks of drug administration, all rats were fasted but allowed to drink water for 12 hours, and fasting tail vein blood was collected. Fasting blood glucose (FBG) was measured using a blood glucose meter; serum insulin level (FINS) of rats was measured using enzyme-linked immunosorbent assay (ELISA).

[0093] The results of fasting blood glucose (FBG) and serum insulin (FINS) levels in rats are shown in Table 4.

[0094] Table 4

[0095] In Table 4, # and## This indicates that the data in the same column shows significant differences between each group and the model group. # This indicates that P < 0.05. ## This indicates that P < 0.01; △ and △△ This indicates that the data in the same column shows significant differences compared to the group in Example 1. △ This indicates that P < 0.05. △△ This indicates that P < 0.01.

[0096] As shown in Table 4, the traditional Chinese medicine composition of the present invention can significantly reduce fasting blood glucose (FBG) in type 2 diabetic rats, indicating that the traditional Chinese medicine composition of the present invention can effectively control blood glucose levels. Serum insulin levels (FINS) in rats increased after administration, which may be due to the drug improving the function of pancreatic β cells and promoting insulin secretion. Furthermore, it can be seen that the traditional Chinese medicine compositions of Examples 1-5 of the present invention have significantly better effects in reducing FBG and increasing FINS than the comparative examples. The experimental results show that the traditional Chinese medicine composition of the present invention has a significantly better therapeutic effect on type 2 diabetes than conventional traditional Chinese medicine compositions prepared by other components, ratios, or other preparation methods.

[0097] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A traditional Chinese medicine composition, characterized in that, The product is made from the following raw materials in parts by weight: 4-8 parts ginseng stem and leaf saponins, 8-16 parts astragalus polysaccharide, 6-12 parts astragaloside A, 8-16 parts catalpol, 6-12 parts rehmannia glycoside D, 8-16 parts verbascoside, 3-5 parts raspberry polysaccharide, 4-8 parts ellagic acid, 4-8 parts poria polysaccharide, 5-10 parts poria acid, 8-16 parts wolfberry polysaccharide, 40-62 parts schisandra, 20-40 parts yam, 40-62 parts ophiopogon japonicus, 40-62 parts trichosanthes kirilowii and 40-62 parts alisma plantago-aquatica.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, Based on weight, it is made from the following raw materials: 5-7 parts ginseng stem and leaf saponins, 10-14 parts astragalus polysaccharide, 8-11 parts astragaloside A, 10-14 parts catalpol, 7-10 parts rehmannia glycoside D, 10-14 parts verbascoside, 3.5-4.5 parts raspberry polysaccharide, 5-7 parts ellagic acid, 5-7 parts poria polysaccharide, 6-9 parts poria acid, 10-14 parts wolfberry polysaccharide, 50-60 parts schisandra, 25-35 parts yam, 45-55 parts ophiopogon japonicus, 45-55 parts trichosanthes kirilowii and 45-55 parts alisma plantago-aquatica.

3. The traditional Chinese medicine composition according to claim 2, characterized in that, The product is made from the following ingredients by weight: 6 parts ginseng stem and leaf saponins, 12 parts astragalus polysaccharide, 10 parts astragaloside A, 12 parts catalpol, 8 parts rehmannia glycoside D, 12 parts verbascoside, 4 parts raspberry polysaccharide, 6 parts ellagic acid, 6 parts poria polysaccharide, 8 parts poria acid, 12 parts wolfberry polysaccharide, 55 parts schisandra, 30 parts yam, 50 parts ophiopogon japonicus, 50 parts trichosanthes kirilowii and 50 parts alisma plantago-aquatica.

4. The method for preparing the traditional Chinese medicine composition according to any one of claims 1-3, characterized in that, Including the following steps: (1) Schisandra chinensis, Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica are pulverized, dried, and then pretreated with a high-voltage pulse electric field to obtain a pretreated mixture; The pretreated mixture is filtered to obtain filtrate A and filter residue A. (2) Filter residue A was initially extracted with ethanol with a volume concentration of 20%-40%, filtered, and filtrate B and filter residue B were obtained. (3) Filter residue B is extracted a second time with ethanol of 60%-80% by volume, filtered, and filtrate C and filter residue C are obtained. (4) Filtrate A, Filtrate B and Filtrate C are dried and mixed to obtain a Chinese medicine extract; then it is mixed with ginseng stem and leaf saponins, astragalus polysaccharide, astragaloside A, catalpol, rehmannia glycoside D, verbascoside, raspberry polysaccharide, ellagic acid, poria polysaccharide, poria acid and wolfberry polysaccharide to obtain a Chinese medicine composition.

5. The preparation method according to claim 4, characterized in that, In step (1), the crushing is performed to crush the material into coarse powder with a particle size of 150-500 μm; In step (1), the drying process involves a drying temperature ≤ 60℃ and a drying time ≤ 48h. In step (1), the conditions for pretreatment by the high-voltage pulse electric field are: electric field strength 20-28kV / cm, number of pulses 5-7, material-liquid ratio 1g:15-25mL, and pH value 7-9.

6. The preparation method according to claim 4, characterized in that, In step (2), the volume concentration of the ethanol is 30%; In step (2), the initial extraction conditions are: extraction temperature 65-75℃, extraction time 2-4h, extraction material-liquid ratio 1g:25-35mL, and the weight of the material in the material-liquid ratio is the total weight of Schisandra chinensis, Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica in step (1).

7. The preparation method according to claim 4, characterized in that, In step (3), the volume concentration of the ethanol is 70%; In step (3), the conditions for the secondary extraction are: extraction temperature 50-60℃, extraction time 1-3h, extraction material-liquid ratio 1g:25-35mL, and the weight of the material in the material-liquid ratio is the total weight of Schisandra chinensis, Dioscorea opposita, Ophiopogon japonicus, Trichosanthes kirilowii and Alisma plantago-aquatica in step (1).

8. The preparation method according to claim 4, characterized in that, In step (4), the drying process involves a drying temperature of ≤60℃ and a drying time of ≤48h.

9. The use of the traditional Chinese medicine composition according to any one of claims 1-3 in the preparation of hypoglycemic products.

10. The application according to claim 9, characterized in that, The product used in this application is a drug.

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