A Chinese medicine composition and its use in preparing a medicine for preventing / treating type 2 diabetes liver damage

Abstract

The present invention discloses a traditional Chinese medicine composition and its use in preparing a medicament for preventing / treating type 2 diabetic liver damage, belonging to the field of biomedicine. The composition comprises, by weight, 10-16 parts of loofah extract, 8-12 parts of papaya powder, 8-12 parts of coix seed powder, 1-8 parts of grape powder, 1-8 parts of kudzu root powder, 1-8 parts of corn silk powder, 1-8 parts of hemp seed powder, 1-8 parts of seaweed powder, 1-8 parts of kelp extract, 1-8 parts of purslane powder, 1-8 parts of lotus leaf powder, 1-8 parts of mint powder, 1-8 parts of angelica powder, 1-8 parts of tangerine peel extract, 1-8 parts of cinnamon powder, 1-8 parts of green tea powder, 4-10 parts of dandelion powder, 4-10 parts of poria powder, and 4-10 parts of hawthorn powder. The traditional Chinese medicine composition can prevent / treat type 2 diabetic liver damage, and solves the problems of many side effects and other problems existing in traditional Chinese medicine compositions for preventing and treating diabetic liver damage.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to a traditional Chinese medicine composition and its application in preparing a medicine for preventing / treating type 2 diabetes liver damage. Background Art

[0002] Type 2 diabetes liver damage is a chronic complication characterized by abnormalities in liver tissue or function that occur during the development and treatment of type 2 diabetes. The liver is a key organ for glucose and lipid metabolism in the human body. During the onset of type 2 diabetes, impaired glucose and lipid metabolism leads to excessive accumulation of glycogen and fat in the liver, causing liver damage. Liver damage further aggravates glucose and lipid metabolism, accelerating the progression of diabetes, creating a vicious cycle of interaction between the two. In recent years, there have been no effective treatments for type 2 diabetes liver damage. Therefore, the search for safe and effective edible and medicinal treatments is of great practical significance.

[0003] Patent CN112807438A discloses a combination drug composition for type 2 diabetes and liver damage and its use. The combination drug composition includes selenium-rich probiotics and the thiazolidinedione drug pioglitazone. Patent CN115251388A discloses a blood sugar-lowering auxiliary composition and its food supplement. The raw materials of the composition include corn silk, kudzu root, prunella vulgaris, cinnamon bark, matsutake mushroom, maitake mushroom, centella asiatica, and schisandra chinensis. Patent CN115944706A discloses a Chinese herbal probiotic complex with blood sugar-lowering function and its preparation method. The complex is prepared by fermenting a fermented chickpea extract and an enzymatically hydrolyzed bitter melon polypeptide with multiple probiotics, performing a Maillard reaction, and then centrifuging and freeze-drying. Patent CN200810196876A relates to a new medical use of tiopronin or its sodium salt, namely, its use in the preparation of a drug for treating liver dysfunction secondary to cerebral ischemia, diabetes, and obesity. Patent CN116058504A discloses a synergistic hypoglycemic composition and a hypoglycemic health drink. The invention uses blue loquat, polygonatum and chaga as the main raw materials, and finally develops a new hypoglycemic health product through animal experiments. In general, these methods have their own characteristics, but there are certain limitations: (1) Many side effects. Most compositions do not consider the adverse effects caused by patients after taking them in the process of treating type 2 diabetes and liver damage. Some methods directly use thiazolidinediones such as pioglitazone, which can easily induce obesity and increase the risk of cardiovascular disease in patients; some methods use sulfaprotinin to treat diabetic liver damage, which can easily cause agranulocytosis, aplastic anemia, thrombocytopenia or other adverse reactions. (2) Poor efficacy. Some methods adjust the taste of the composition by adding sweeteners and food additives such as sucralose, sodium citrate, γ-cyclodextrin and flavors that are not suitable for diabetic patients to consume excessively, which can easily increase the patient's blood sugar and induce diabetic liver damage. (3) Harsh storage environment. Some methods use probiotic complexes such as Lactobacillus acidophilus, Lactobacillus casei, Bifidobacterium selenium-enriched, and Lactobacillus plantarum in the preparation of traditional Chinese medicine compositions. However, the storage environment of these probiotics is relatively harsh and generally requires low-temperature refrigeration. Moreover, after entering the human digestive system, they may be killed by gastric juice, resulting in a small number of surviving bacteria and a short time of staying in the intestines. They are easily excreted from the body, making it impossible to fully exert the efficacy of the medicine. (4) The preparation process is complicated. Some existing technologies have complex preparation processes and cumbersome operations. They require multiple steps such as enzymatic hydrolysis, extraction, and fermentation. The production process is complicated, which is not conducive to the industrial development of the composition. Summary of the Invention

[0004] In order to overcome the shortcomings of the above-mentioned prior art, the purpose of the present invention is to provide a traditional Chinese medicine composition and its use in the preparation of a drug for preventing / treating type 2 diabetic liver damage, so as to solve the technical problems of existing compositions for treating diabetic liver damage, such as multiple side effects, poor efficacy, harsh storage environment and complex preparation process.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions:

[0006] The first aspect of the present invention discloses a traditional Chinese medicine composition, which comprises, by weight, 10 to 16 parts of loofah extract, 8 to 12 parts of papaya powder, 8 to 12 parts of coix seed powder, 1 to 8 parts of grape powder, 1 to 8 parts of kudzu root powder, 1 to 8 parts of corn silk powder, 1 to 8 parts of seaweed powder, 1 to 8 parts of kelp extract, 1 to 8 parts of purslane powder, 1 to 8 parts of lotus leaf powder, 1 to 8 parts of mint powder, 1 to 8 parts of angelica powder, 1 to 8 parts of tangerine peel extract, 1 to 8 parts of cinnamon powder, 1 to 8 parts of green tea powder, 4 to 10 parts of dandelion powder, 1 to 8 parts of hemp seed powder, 4 to 10 parts of poria powder and 4 to 10 parts of hawthorn powder; in the traditional Chinese medicine composition, the valine content does not exceed 4.50%, and the leucine content does not exceed 5.80%.

[0007] Preferably, the ingredients, by weight, include: 14 parts of loofah extract, 10 parts of papaya powder, 10 parts of coix seed powder, 3 parts of grape powder, 3 parts of kudzu root powder, 3 parts of corn silk powder, 3 parts of hemp seed powder, 3 parts of seaweed powder, 3 parts of kelp extract, 3 parts of purslane powder, 3 parts of lotus leaf powder, 3 parts of mint powder, 3 parts of angelica powder, 3 parts of tangerine peel extract, 3 parts of cinnamon powder, 3 parts of green tea powder, 9 parts of dandelion powder, 9 parts of poria powder and 9 parts of hawthorn powder.

[0008] The second aspect of the present invention discloses the use of the above-mentioned traditional Chinese medicine composition in the preparation of a drug for preventing / treating liver damage in type 2 diabetes.

[0009] Preferably, the drug is a drug that inhibits the activity of α-amylase and α-glucosidase.

[0010] Preferably, the drug is a drug that lowers liver cholesterol, triglyceride, low-density lipoprotein cholesterol, and malondialdehyde levels.

[0011] Preferably, the drug is a drug that increases the content of high-density lipoprotein cholesterol and liver glycogen in the liver and the activity of superoxide dismutase and glutathione peroxidase.

[0012] Preferably, the drug is a drug that reduces serum aspartate aminotransferase and alanine aminotransferase levels.

[0013] Preferably, the drug is a drug that alleviates vacuolation, deformation and disordered arrangement of hepatocytes in liver tissue.

[0014] Preferably, the drug is a drug that alleviates pancreatic islet cell vacuolation, fat hyperplasia, interstitial edema and inflammatory cell infiltration.

[0015] Compared with the prior art, the present invention has the following beneficial effects:

[0016] The present invention provides a Chinese medicine composition, which uses medicinal and edible plants and plant foods as main raw materials, and contains medicinal and edible plant components such as loofah extract, grape powder, papaya powder, coix seed powder, dandelion powder, kudzu root powder, corn silk powder, hemp seed powder, seaweed powder, kelp extract, poria powder, hawthorn powder, purslane powder, lotus leaf powder, mint powder, angelica powder, tangerine peel extract, cinnamon powder and green tea powder. (1) The Chinese medicine composition is composed of multiple medicinal and edible plants, does not contain other medicinal excipients or food additives that are harmful to patients with T2DM liver damage, and has mild performance; the active ingredients selected from loofah extract, kelp extract and tangerine peel extract can ensure that each active ingredient is fully released, remove ineffective or harmful ingredients contained in the medicinal materials, have few side effects and are highly safe. (2) The valine content in the Chinese medicine composition does not exceed 4.50%, and the leucine content does not exceed 5.80%, which helps reduce the risk of T2DM liver damage. (3) The kudzu root powder, dandelion powder, purslane powder, hawthorn powder and tangerine peel extract selected in the Chinese medicine composition have lipid-lowering effects, can increase the activity levels of SOD and GSH-Px, and improve T2DM liver damage; furanocoumarins in angelica dahurica can significantly inhibit the activity of aldose reductase, improve diabetic cataracts and diabetes; loofah has the effects of promoting blood circulation, activating collaterals, detoxifying and dispersing nodules, and tuckahoe powder, seaweed powder and hemp seed powder can enhance the effects of loofah; the polysaccharide components in kelp extract can inhibit the activity of α-amylase and α-glucosidase; cinnamon powder has the effect of lowering blood sugar and relieving diabetic syndrome; papaya powder, mint powder, lotus leaf powder and coix seed powder regulate qi and strengthen the spleen, promote water expelling and dampness removal, and play a role in lowering blood sugar and protecting the liver. (4) The Chinese medicine composition is added with components such as green tea powder, grape powder, corn silk powder and seaweed powder, which can reduce the bitterness and fishy smell in the original medicinal materials and effectively improve the taste and flavor of the Chinese medicine composition when taken. (5) The preparation process of the Chinese medicine composition is simple and can fully retain the active ingredients in the composition; the operation is controllable, the storage is convenient, and it is easy to industrialize. Therefore, the Chinese medicine composition can be used as an alternative drug for the clinical treatment of liver damage in patients with T2DM, providing a new therapeutic strategy for the prevention, treatment, or auxiliary treatment of liver damage caused by diabetes. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is a diagram showing the inhibitory effect of the Chinese medicine composition on α-amylase activity;

[0018] Figure 2 This is a diagram showing the inhibitory effect of the Chinese medicine composition on α-glucosidase activity;

[0019] Figure 3 The figure shows the effect of the Chinese herbal medicine composition on the water intake and food intake of T2DM mice; A represents the water intake and B represents the food intake;

[0020] Figure 4 This is a graph showing the effect of the Chinese herbal composition on the body weight of T2DM mice;

[0021] Figure 5 The figure shows the effect of the Chinese herbal composition on oral glucose tolerance in T2DM mice; different superscript letters indicate significant differences among the groups (P<0.05);

[0022] Figure 6 The figure shows the effect of the Chinese herbal medicine composition on the insulin content in T2DM mice; different superscript letters indicate significant differences among the groups (P<0.05);

[0023] Figure 7 The figure shows the effect of the Chinese herbal medicine composition on the insulin sensitivity index of T2DM mice; different superscript letters indicate significant differences among the groups (P<0.05);

[0024] Figure 8 HE×200 images of liver morphology of T2DM mice; A: NC group, B: MC group, C: PC group, D: S 1500 Group, E:S 1000 Group, F:S 500 Group;

[0025] Figure 9 HE×200 images of pancreatic morphology of T2DM mice; A: NC group, B: MC group, C: PC group, D: S 1500 Group, E:S 1000 Group, F:S 500 Group. DETAILED DESCRIPTION

[0026] To facilitate understanding of the features and effects of the present invention by those skilled in the art, the following provides a general description and definition of the terms and expressions used in the specification and claims. Unless otherwise indicated, all technical and scientific terms used herein have the ordinary meanings as understood by those skilled in the art regarding the present invention. In the event of conflict, the definitions in this specification shall prevail.

[0027] The theories or mechanisms described and disclosed herein, whether correct or incorrect, should not limit the scope of the present invention in any way, that is, the present invention can be implemented without being limited by any specific theory or mechanism.

[0028] Herein, all features such as values, amounts, amounts, and concentrations defined in numerical ranges or percentage ranges are for brevity and convenience only. Accordingly, the description of numerical ranges or percentage ranges should be considered to include and specifically disclose all possible subranges and individual values ​​within the range (including integers and fractions).

[0029] In this document, unless otherwise specified, “include,” “including,” “contains,” “has” or similar terms cover the meanings of “consisting of” and “mainly consisting of,” for example, “A includes a” covers the meanings of “A includes a and other” and “A only includes a.”

[0030] In this document, for the sake of brevity, not all possible combinations of the various technical features in each embodiment or example are described. Therefore, as long as there are no contradictions in the combination of these technical features, the various technical features in each embodiment or example can be combined in any way, and all possible combinations should be considered to be within the scope of this specification.

[0031] Through the inventor's own clinical practice research of traditional Chinese medicine, it was found that many patients with T2DM, metabolic disorder-type obesity, hyperglycemia and hyperlipidemia have "triple burner" pathological problems. Therefore, under the guidance of the "triple burner" theory of traditional Chinese medicine, after years of clinical experience and experiments, a traditional Chinese medicine composition is provided, which includes, by weight: 10-16 parts of loofah extract, 8-12 parts of papaya powder, 8-12 parts of coix seed powder, 1-8 parts of grape powder, 1-8 parts of kudzu root powder, 1-8 parts of corn silk powder, 1-8 parts of hemp seed powder, 1-8 parts of seaweed powder, 1-8 parts of kelp extract, 1-8 parts of purslane powder, 1-8 parts of lotus leaf powder, 1-8 parts of mint powder, 1-8 parts of angelica powder, 1-8 parts of tangerine peel extract, 1-8 parts of cinnamon powder, 1-8 parts of green tea powder, 4-10 parts of dandelion powder, 4-10 parts of poria powder and 4-10 parts of hawthorn powder, for alleviating T2DM liver damage.

[0032] Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiments are only used to illustrate the present invention and are not used in limiting the scope of the present invention.In addition, should be understood that after reading the content taught by the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms fall equally within the scope limited by the appended claims of the application.

[0033] The following examples were prepared using conventional instruments and equipment in the art. Experimental methods in the following examples, where specific conditions are not specified, were generally performed under conventional conditions or according to the conditions recommended by the manufacturer. The various raw materials used in the following examples, unless otherwise specified, were conventional commercially available products, with specifications conventional in the art. In the present specification and the following examples, unless otherwise specified, "%" indicates percentage by weight, "part" indicates parts by weight, and "ratio" indicates weight ratio.

[0034] Table 1 Formula of Chinese medicine composition that helps prevent / treat liver damage in type II diabetes

[0035]

[0036]

[0037] According to the formula shown in Table 1, the Chinese medicine compositions of Examples 1 to 9 were prepared. In these Chinese medicine compositions, the valine content did not exceed 4.50%, and the leucine content did not exceed 5.80%.

[0038] The above Chinese medicine composition or any one of the embodiments can be prepared according to the following method:

[0039] Step 1: Prepare and sterilize the raw materials of each component, including loofah, papaya, coix seed, grape, kudzu root, corn silk, hemp seed, seaweed, kelp, purslane, lotus leaf, mint, angelica dahurica, tangerine peel, cinnamon, green tea, dandelion, poria and hawthorn;

[0040] Step 2: Grind and sieve the sterilized raw materials separately to obtain powders of each component with a particle size of 800 mesh;

[0041] Step 3: Mix the loofah powder, tangerine peel powder, and kelp powder with double-distilled water in a solid-liquid ratio of 1:3, soak for 20 minutes, decoct for 20-30 minutes, and filter to obtain aqueous extracts of each component; repeat the decoction 1-2 times, combine the components, and filter the aqueous extracts obtained;

[0042] Step 4: freeze-drying the aqueous extracts of the components at a temperature of -60±3°C and a vacuum degree of 15±3Pa for 20 to 24 hours, and grinding the extracts to obtain loofah powder extract, tangerine peel powder extract, and kelp extract, respectively;

[0043] Step 5: Thoroughly mix the above raw materials according to their weight proportions, and determine the valine and leucine contents in the traditional Chinese medicine composition. If the valine content in the traditional Chinese medicine composition does not exceed 4.50%, and the leucine content does not exceed 5.80%, the traditional Chinese medicine composition is prepared. If the valine content in the traditional Chinese medicine composition exceeds 4.50%, and / or the leucine content exceeds 5.80%, readjust the specific proportions of each component in the traditional Chinese medicine composition until the valine and leucine contents meet the requirements.

[0044] The Chinese medicine compositions of the present invention are further illustrated below through test examples. The Chinese medicine compositions used in the tests are all prepared according to the optimal ratio Example 8.

[0045] 1. Amino acid composition analysis of traditional Chinese medicine compositions

[0046] 1. Experimental Methods

[0047] The amino acid composition of the Chinese medicine composition was determined by referring to the laboratory method (Chemical composition, antioxidant activity and protective effect of organic selenium-rich matcha on DNA damage [J]. Journal of Zhejiang Agricultural Sciences, 2023, 35(9): 2222-2232), which is briefly described as follows: 0.2 g of the Chinese medicine composition in the above embodiment was weighed and dissolved in 5 mL of 6.0 mol / L HCl solution. After ultrasonic treatment for 2 min, the mixture was placed in a CEMMARS6 microwave digester at 180°C for digestion; after 30 min, the digestion solution was taken out and diluted to 50 mL with double distilled water. 1 to 3 mL of the solution was drawn from the diluted solution and added to an equal volume of 0.5 mol / L NaOH. The solution was redivided to 10 mL with double distilled water and filtered. The filtrate was taken and subjected to amino acid determination in an L-8900 amino acid analyzer.

[0048] 2. Experimental Results

[0049] Abnormal amino acid metabolism is associated with the occurrence of metabolic diseases such as obesity, insulin resistance, and T2DM. Studies have found that high levels of leucine and valine can promote insulin resistance by interfering with the insulin signaling pathway or directly inhibiting glucose transport and phosphorylation in skeletal muscle, thereby increasing the risk of T2DM; while high levels of glycine can promote glutathione synthesis and inhibit the production of intracellular reactive oxygen species to protect pancreatic β cells from oxidative stress damage, thereby promoting insulin secretion and reducing the risk of T2DM. As shown in Tables 2 and 3, the Chinese medicine composition of the present invention contains 17 amino acids, and essential amino acids account for 28.21% of the total amino acids. According to the comparative analysis of the amino acid score (AAS) and chemical score (CS) models, valine is the first limiting amino acid in the Chinese medicine composition, with AAS and CS values ​​of 0.59 and 0.44, respectively; leucine is the second limiting amino acid in the Chinese medicine composition, with AAS and CS values ​​of 0.64 and 0.47, respectively. In summary, the traditional Chinese medicine composition has low leucine and valine contents and high glycine content, which helps to reduce the risk of T2DM and maintain the body's liver at a healthy level.

[0050] Table 2 Amino acid composition of Chinese medicine composition

[0051]

[0052] Table 3 Comparative analysis of AAS and CS of traditional Chinese medicine compositions

[0053]

[0054]

[0055] 2. Study on the inhibitory effect of Chinese herbal medicine combination on α-amylase and α-glucosidase activities

[0056] 1. Test article

[0057] Before use, the Chinese medicine composition of Example 8 was prepared into test solutions of different concentrations (0.5-4.0 mg / mL) using PBS (pH 6.9, 20 mM).

[0058] 2. Experimental Methods

[0059] (1) Referring to the laboratory method (Effects of Selenium-enriched Chrysanthemum on the Expression of Genes Related to Drosophila Lifespan and Antioxidant Defense and the Development of Quality Standards [D]. Shaanxi Normal University, 2019), the inhibitory effect of the Chinese herbal composition on α-amylase activity was determined. The results are as follows: 200 μL of the Chinese herbal composition solution of different concentrations was mixed evenly with 50 μL of α-amylase solution, incubated in a 25°C water bath for 30 minutes, and then 500 μL of 1% starch solution was added and placed in a 40°C water bath for 20 minutes. Finally, 500 μL of dinitrosalicylic acid color reagent was added to terminate the above reaction, and then placed in a boiling water bath for 10 minutes, cooled to room temperature, and the volume was fixed and the absorbance at 540 nm was measured. In the experiment, double distilled water was used to replace the enzyme solution as the sample blank group, double distilled water was used to replace the Chinese herbal composition solution as the control group, and double distilled water was used to replace the Chinese herbal composition solution and enzyme solution as the control blank group. Acarbose was used as the positive control. The operation steps were the same as above. The experiment was repeated 3 times and the average value was taken. 50 The value is calculated as follows:

[0060] α-amylase inhibition rate (%) = (1-sample △ A540 / Control △A540 )*100%

[0061] Where: Sample △ A540 The difference between the absorbance of the sample group and the absorbance of the blank group is the difference between the absorbance of the sample group and the blank group. A540 It is the difference between the absorbance value of the control group and the absorbance value of the blank group.

[0062] (2) The inhibitory effect of the Chinese herbal composition on α-glucosidase activity was determined by the reference test method (Inhibitory activity of α-amylase and α-glucosidase byplant extracts from the Brazilian cerrado [J]. Planta Med, 2012, 78(4): 393-9). The results are as follows: 100 μL of the Chinese herbal composition solution of different concentrations and p-nitrophenyl-α-D-pyranoglucoside (5 mM, 0.1 mL) were mixed and incubated in a 37°C water bath for 10 min. 200 μL of α-glucosidase (0.1 U / mL) was then added and the reaction continued for 30 min. The absorbance at 405 nm was finally measured. In the experiment, double-distilled water was used to replace the enzyme solution as the sample blank group, double-distilled water was used to replace the Chinese herbal composition solution as the control blank group, and double-distilled water was used to replace the Chinese herbal composition solution and the enzyme solution as the control blank group. Acarbose was used as the positive control. The operation steps were the same as above.

[0063] The experiment was repeated 3 times and the average value was taken. The inhibition rate and IC50 The value is calculated as follows:

[0064] α-glucosidase inhibition rate (%) = (1-sample △ A405 / Control△ A405 )*100%

[0065] Where: Sample △ A405 The difference between the absorbance of the sample group and the absorbance of the blank group is the difference between the absorbance of the sample group and the blank group. A405 It is the difference between the absorbance value of the control group and the absorbance value of the blank group.

[0066] 3. Experimental Results

[0067] Depend on Figure 1 and Figure 2 It can be seen that the Chinese medicine composition has a certain inhibitory effect on the activity of α-amylase and α-glucosidase, showing a clear dose-dependent effect. In the concentration range of 0.5-4.0 mg / mL, as the concentration of the Chinese medicine composition increases, the inhibition rate of α-amylase and α-glucosidase also gradually increases, and both have a good linear correlation (R 2 =0.9983, R 2 =0.9916). As shown in Table 4, compared with the inhibitory effect of acarbose on the activities of the two enzymes, the Chinese medicine composition can inhibit the activities of α-amylase and α-glucose, hinder the decomposition of starch and maltose, delay the body's digestion and absorption of carbohydrates, reduce endogenous glucose production, and help alleviate hyperglycemia symptoms.

[0068] Table 4 Fitting equations and IC values ​​for the inhibitory effects of the Chinese herbal composition on α-amylase and α-glucosidase 50

[0069]

[0070]

[0071] 3. Human Trial Application Research of Chinese Medicine Composition

[0072] 1. Research Methods

[0073] According to the regulations and principles of population tasting, 3 volunteers with abnormal blood sugar were selected to take the above-mentioned Chinese medicine combination, 3 doses per day, for 1 month.

[0074] 2. Research Results and Analysis

[0075] After taking the Chinese medicine composition for one month, the weight loss trend of the three volunteers with abnormal blood sugar was alleviated to a certain extent. The test results of blood sugar and blood pressure are shown in Table 5. The blood sugar and blood pressure levels of the volunteers with abnormal blood sugar dropped significantly, approaching normal levels, and no symptoms of diabetic liver damage were observed.

[0076] Table 5 Blood sugar and blood pressure test results

[0077]

[0078] IV. Study on the intervention effect of Chinese herbal medicine combination on diabetic liver damage in experimental animals

[0079] In order to further confirm the effect of the above-mentioned Chinese medicine composition on diabetes and its liver damage complications and reveal its mechanism of intervention in diabetic liver damage, an animal experimental system was used to study the hypoglycemic and hypolipidemic effects of the Chinese medicine composition on mice and its effects and mechanisms on diabetic liver damage.

[0080] (1) Study on the hypoglycemic effect of Chinese herbal medicine combination on mice

[0081] 1. Animals

[0082] Kunming mice (80, 4-5 weeks old, body weight 16-20 g, male) were purchased from Sibeifu (Beijing) Biotechnology Co., Ltd., production certificate number: SCXK (Beijing) 2019-0010.

[0083] 2. Experimental Methods

[0084] (1) Preparation of T2DM mouse model and group administration experiment: All Kunming mice were fed with normal experimental mouse feed for one week and then randomly divided into experimental group and normal group (NC: 10 mice); the experimental group was fed with high-fat and high-sugar feed, and the normal group was fed with normal feed for 5 weeks. All experimental mice were fasted but not watered for 12 hours (overnight), and their fasting blood glucose was measured and weighed. Then, the experimental group mice were intraperitoneally injected with 95 mg / kg bw (Body Weight) of Streptozotocin (STZ) (prepared immediately), and the normal group mice were injected with an equal volume of normal saline. After 72 hours, they were fasted for 12 hours, and their fasting blood glucose (FBG) was measured. FBG ≥ 11.0 mmol·L -1 The mice were determined to be T2DM mice. The mice with T2DM model were randomly divided into 5 groups again, namely model group (MC), glimepiride positive control group (PC), 500, 1000, 1500 mg / kg composition group (S 500 、S 1000 、S 1500Each group consisted of 10 mice. The glimepiride positive control group and the combination group were treated with the corresponding drugs, while the NC and MC groups were administered with an equal volume of normal saline for four consecutive weeks. During the dosing period, mice in all groups except the NC group continued to be fed a high-fat, high-sugar diet. The mice's coat, mental state, and urine output were observed during the experiment. The average body weight, food intake (food intake = added food - remaining food), and water intake (water intake = added water - remaining water) of each group were recorded every three days. Fasting blood glucose levels were measured every other week.

[0085] (2) Mouse glucose tolerance test: The experimental mice were fed with the drug for 4 weeks and fasted for 12 hours without water. The fasting blood glucose values ​​of the mice in each group were measured (0 hour). Then, the mice in the control group and the combination group were gavaged with glimepiride and different doses of the Chinese medicine combination. The NC and MC groups were given normal saline. After 20 minutes, the mice in each group were again gavaged with 2 g·kg -1 bw glucose solution, and finally measure the blood glucose levels of mice at 0.5h and 2h after administration of glucose, and calculate the area under the blood glucose curve (AUC) according to the following formula.

[0086] AUC=[(0h blood glucose+0.5h blood glucose)*0.5] / 2+[(0.5h blood glucose+2h blood glucose)*1.5] / 2

[0087] (3) Study on the degree of insulin resistance in mice: The mouse model was prepared and grouped according to the above method. The serum of mice was collected and the INS content in the serum was determined according to the method in the kit instructions. The degree of insulin resistance was evaluated using the homeostasis model assessment-insulin resistance index (HOMA-IR). The calculation formula is as follows: HOMA-IR = (FBG mmol L -1 *INS mIU L -1 ) / 22.5

[0088] (4) Study on antioxidant capacity in mice: Mouse models were prepared and grouped according to the above method. Serum and liver tissue were collected from mice, and the levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA) in serum and liver tissue were determined according to the kit instructions.

[0089] 3. Statistical Analysis: All data were analyzed using SPSS 26.0 software. Results are presented as mean ± standard deviation and statistically analyzed. One-way analysis of variance was used for data comparisons. p < 0.05 and p < 0.01 were considered statistically significant, highly significant, or statistically insignificant.

[0090] 4. Experimental Results

[0091] (1) Analysis of the physical signs, eating and drinking conditions of T2DM mice: As shown in Table 6, the study found that compared with the NC group mice, the fasting blood glucose level of T2DM model mice was higher, the hair of the MC group mice was dark yellow and sparse, and the drinking and eating conditions gradually increased ( Figure 3 ), weight loss ( Figure 4 ), the litter was damp and had a noticeable odor; after artificial intervention with different doses of Chinese medicine composition and glimepiride, the composition group S 500 、S 1000 With S 1500 The mice's weight increased in the later stages of treatment, their water and food intake decreased significantly, and their physical appearance improved significantly. In particular, the high-dose group showed a significant decrease in FBG values ​​by 32.13% (Table 6). This suggests that the Chinese herbal composition can indeed improve the physical appearance and the typical "three mores and one less" symptoms of T2DM mice.

[0092] Table 6 Effect of Chinese herbal composition on fasting blood glucose (FBG) in T2DM mice

[0093]

[0094] *P<0.05**P<0.01 compared with MC group; different superscript letters indicate significant differences among the groups (P<0.05).

[0095] (2) Analysis of glucose tolerance test in T2DM mice: Improving insulin sensitivity is an important starting point for treating T2DM, and glucose tolerance is one of the important indicators for evaluating insulin sensitivity. As shown in Table 7, compared with the NC group, the blood glucose levels of the mice in the MC group, PC group, and combination group reached a peak at 0.5 h after glucose loading; when glimepiride and the Chinese medicine combination were given, the PC group and S group showed a peak at 0.5 h. 1000 、S 1500 The blood glucose level of mice in the MC group decreased significantly after 2 hours compared with that in the MC group (P<0.05). Figure 5 As shown in the figure, compared with the NC group, the area under the AUC curve of the MC group was significantly increased (P<0.05); after artificial intervention, the S 1000 、S 1500 Compared with glimepiride, the area under the AUC curve of T2DM mice was significantly reduced (P<0.05), indicating that the Chinese medicine composition can alleviate the glucose load of T2DM mice and improve the glucose tolerance of T2DM mice.

[0096] Table 7 Effects of Chinese herbal composition on glucose tolerance in T2DM mice

[0097]

[0098] Note: Different superscript letters indicate significant differences among the groups (P<0.05).

[0099] (3) Analysis of insulin resistance in T2DM mice: T2DM is non-insulin-dependent diabetes, and insulin is the only hormone in the body that can lower blood sugar. The pancreatic islet cells in T2DM patients can still produce insulin, but the insulin produced cannot meet the normal needs of the body and the sensitivity is reduced, which usually manifests as insulin resistance. HOMA-IR is often used to evaluate the degree of insulin resistance. Figure 6 and Figure 7 As shown in the results, compared with the NC group, the INS content of the mice in the MC group was significantly increased (P < 0.01), and the HOMA-IR index was also significantly increased (P < 0.01), indicating insulin resistance. After intervention with glimepiride and the Chinese medicine composition, the INS content and HOMA-IR index of the mice in the PC group and the composition group were reduced to varying degrees compared with the MC group. This indicates that the PC group and the composition S 1500 The INS content and HOMA-IR index of T2DM mice were significantly reduced in both groups (P<0.01).

[0100] (4) Analysis of oxidative stress response in mice: Hyperglycemia and hyperlipidemia can both lead to oxidative stress, and oxidative stress is involved in impaired pancreatic B cell function and insulin resistance, thereby promoting the occurrence of diabetic complications; the level of SOD and GSH-Px activity reflects the body's ability to scavenge free radicals, and MDA indirectly reflects the degree to which the body's cells are attacked by free radicals. The present invention uses diabetic mice induced by high fat and high sugar combined with STZ as a model to explore whether the Chinese medicine composition has antioxidant properties. The results are shown in Table 8: The activities of SOD and GSH-Px in mice in the MC group were significantly lower than those in the NC group, and the MDA level was significantly higher than that in the NC group (P < 0.05); after treatment with glimepiride and the Chinese medicine composition, the activities of SOD and GSH-Px gradually increased, and the MDA level decreased to varying degrees. Composition S 1500 The activity of SOD and GSH-Px in the control group was significantly increased (P < 0.01), and MDA accumulation was reduced. This indicates that the Chinese medicine composition can alleviate the oxidative stress level of T2DM mice by increasing the activity of antioxidant enzymes SOD and GSH-Px and reducing MDA accumulation.

[0101] Table 8 Effects of Chinese herbal medicine combination on antioxidant enzymes and MDA in T2DM mice

[0102]

[0103] Note: Different superscript letters indicate significant differences among the groups (P<0.05).

[0104] (II) Study on the effect of Chinese herbal medicine combination on diabetic liver damage in mice

[0105] 1. Animals: Same as above.

[0106] 2. Experimental Methods

[0107] (1) The mouse model was prepared and grouped according to the above method. Liver tissue and serum were obtained from the mice, and the liver glycogen content, aspartate aminotransferase (AST), and alanine aminotransferase (ALT) levels were determined according to the kit instructions. The statistical analysis method was the same as above.

[0108] (2) Study on pathological sections of mouse organ tissues: The liver and pancreas tissues of mice were fixed with 4% paraformaldehyde solution for 72 hours, and then dehydrated with different concentrations of ethanol solution. After drying, xylene clearing, wax immersion, embedding, sectioning and drying, HE staining was performed. The tissue pathology was observed under a 200x light microscope and photographed.

[0109] 3. Experimental Results

[0110] (1) Analysis of liver function in T2DM mice: The liver is one of the most important organs for glucose metabolism and can convert glucose into glycogen for storage through various pathways, thus restoring blood sugar to normal levels. As shown in Table 9, compared with the NC group, the amount of glycogen in the liver of the MC group mice was significantly reduced (P ≤ 0.05); after intervention with the Chinese medicine composition and glimepiride, the composition S 1500 The liver glycogen content in mice in the PC group was significantly increased (P≤0.05), and the S 500 and S 1000 There was no significant difference in the amount of liver glycogen in the mice of the MC group and the NC group (P≤0.05), indicating that the Chinese medicine composition can indeed promote glucose consumption and inhibit gluconeogenesis. ALT and AST are important indicators for evaluating liver function. As shown in Table 9, compared with the NC group, the AST and ALT activities of the MC group were significantly increased (P<0.01); after artificial intervention treatment, the composition S 1000 With S 1500 It significantly reduced AST and ALT in the serum of mice (P<0.05) and effectively alleviated liver damage caused by T2DM.

[0111] Table 9 Effects of Chinese herbal medicine combination on liver glycogen, AST and ALT levels in T2DM mice liver

[0112]

[0113] Note: Different superscript letters on the right side of the numbers in the same column indicate significant differences in content among the groups (P<0.05).

[0114] (2) Analysis of mouse liver pathological sections: Figure 8As shown in the figure, the liver lobule structure of the NC group mice was clear, the cytoplasm of the liver cells was uniform, and there was no cell deformation; the liver tissue structure of the MC group mice was disordered, the cells were arranged in disorder, the outline was blurred, some were edematous and even had vacuoles and deformation, the cell nuclei were unevenly arranged and even aggregated, and the cytoplasm was fused. 1500 The liver tissue of mice treated with Glemepiride tended to be normal, with less vacuolar deformation and more regular arrangement of liver cells, which was basically close to the effect of the positive control group Glemepiride.

[0115] (3) Analysis of mouse pancreatic pathological sections: Figure 9 As shown in the figure, the pancreatic islet structure of the NC group mice was clear, the islet cells were normal in morphology and evenly distributed; the pancreatic islets of the MC group mice were wrapped by a large amount of lipids, with more vacuoles in the pancreas, and obvious fat hyperplasia, interstitial edema and inflammatory cell infiltration. 1500 The number of pancreatic cells gradually increased, the islet structure was clearer, and the degree of fat hyperplasia, interstitial edema and inflammatory cell damage was better.

[0116] (III) Study on the effect of Chinese herbal medicine combination on liver lipids in mice

[0117] 1. Animals: Same as above

[0118] 2. Experimental Methods

[0119] Mouse models were prepared and grouped as described above. Serum and liver tissue were collected and the levels of high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), cholesterol (TC), and triglyceride (TG) in serum and liver were measured according to the kit instructions. Statistical analysis was performed as described above.

[0120] 3. Experimental Results

[0121] "Lipotoxicity" is also an important factor in the pathogenesis of diabetes. High blood lipids can also increase insulin resistance, thereby worsening blood sugar. As shown in Table 10, in order to explore the effect of the Chinese medicine composition on lipid metabolism in diabetic mice, this study detected the blood lipid levels in the mouse serum. The study observed that compared with the NC group, the MC group mice showed obvious symptoms of lipid metabolism disorders, specifically, the TG, TC, and LDL-C levels in the serum were significantly increased, and the HDL-C content was significantly decreased (P < 0.05); after drug intervention, the PC group and the Chinese medicine composition group S 1000 With S 1500 The HDL-C content in the serum of mice in the dosage group was significantly increased, while the TC, TG and LDL-C levels were significantly decreased (P < 0.05), and there was a significant statistical difference (P < 0.05), indicating that the Chinese medicine composition can alleviate the lipid metabolism disorder in diabetic mice to a certain extent.

[0122] Table 10 Effects of Chinese herbal composition on blood lipids in T2DM mice

[0123]

[0124] The above content is only for explaining the technical idea of ​​the present invention and cannot be used to limit the protection scope of the present invention. Any changes made on the basis of the technical solution in accordance with the technical idea proposed by the present invention shall fall within the protection scope of the claims of the present invention.

Claims

1. A Chinese medicine composition, characterized in that The Chinese medicine composition comprises, by weight, 10 to 16 parts of loofah extract, 8 to 12 parts of papaya powder, 8 to 12 parts of coix seed powder, 1 to 8 parts of grape powder, 1 to 8 parts of kudzu root powder, 1 to 8 parts of corn silk powder, 1 to 8 parts of seaweed powder, 1 to 8 parts of kelp extract, 1 to 8 parts of purslane powder, 1 to 8 parts of lotus leaf powder, 1 to 8 parts of mint powder, 1 to 8 parts of angelica powder, 1 to 8 parts of tangerine peel extract, 1 to 8 parts of cinnamon powder, 1 to 8 parts of green tea powder, 4 to 10 parts of dandelion powder, 1 to 8 parts of hemp seed powder, 4 to 10 parts of poria powder and 4 to 10 parts of hawthorn powder; in the Chinese medicine composition, the valine content does not exceed 4.50%, and the leucine content does not exceed 5.80%.

2. A Chinese medicine composition according to claim 1, characterized in that, By weight, it consists of 14 parts of loofah extract, 10 parts of papaya powder, 10 parts of coix seed powder, 3 parts of grape powder, 3 parts of kudzu root powder, 3 parts of corn silk powder, 3 parts of hemp seed powder, 3 parts of seaweed powder, 3 parts of kelp extract, 3 parts of purslane powder, 3 parts of lotus leaf powder, 3 parts of mint powder, 3 parts of angelica powder, 3 parts of tangerine peel extract, 3 parts of cinnamon powder, 3 parts of green tea powder, 9 parts of dandelion powder, 9 parts of poria powder and 9 parts of hawthorn powder.

3. Use of a traditional Chinese medicine composition according to claim 1 or 2 in the preparation of a medicament for preventing / treating liver damage in type 2 diabetes.

4. The use according to claim 3, characterized in that The drug is a drug that inhibits the activities of α-amylase and α-glucosidase.

5. The use according to claim 3, characterized in that The drug is a drug that lowers liver cholesterol, triglyceride, low-density lipoprotein cholesterol or malondialdehyde levels.

6. The use according to claim 3, characterized in that The medicine is a medicine for increasing the content of high-density lipoprotein cholesterol and liver glycogen in the liver and the activity of superoxide dismutase and glutathione peroxidase.

7. The use according to claim 3, characterized in that The drug is a drug for lowering serum aspartate aminotransferase and alanine aminotransferase levels.

8. The use according to claim 3, characterized in that The medicine is a medicine for alleviating vacuolation, deformation and disordered arrangement of hepatocytes in liver tissue.

9. The use according to claim 3, characterized in that The medicine is a medicine for alleviating pancreatic islet cell vacuolation, fat hyperplasia, interstitial edema and inflammatory cell infiltration.

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