A traditional Chinese medicine composition for preventing and treating metabolic-related fatty liver disease combined with type 2 diabetes and application thereof

Abstract

The application provides a traditional Chinese medicine composition for preventing and treating metabolic-related fatty liver disease combined with type 2 diabetes and application thereof, and the traditional Chinese medicine composition is made of raw materials in the following weight parts: 6-30 parts of Huangqi, 0-20 parts of Ganjiang, 4-20 parts of Shaoyao, 4-20 parts of Gancao and 2-30 parts of Dazao. The medicine provided by the application can simultaneously significantly improve the abnormal glucose metabolism and liver metabolism damage of a "sugar-liver comorbidity" model animal, realizes integrated intervention on a core pathological link of the disease, can produce an unexpected integrated treatment effect on the "sugar-liver comorbidity", significantly reduces adverse reactions of diarrhea, improves gastrointestinal tolerance, is more suitable for the core pathogenesis of the "sugar-liver comorbidity" with cold and heat mixed together, and is beneficial to improving long-term medication safety.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, specifically to a traditional Chinese medicine composition and its application in the preparation of a drug for the prevention and / or treatment of metabolic-related fatty liver disease complicated with type 2 diabetes (hereinafter referred to as "diabetic liver comorbidity"). Background Technology

[0002] Metabolic fatty liver disease and type 2 diabetes are common metabolic disorders that often coexist, mutually reinforcing each other and creating a vicious cycle, clinically known as "diabetic-hepatic comorbidity." Currently, modern medicine lacks specific drugs for "diabetic-hepatic comorbidity," and often employs a comprehensive treatment plan combining hypoglycemic agents with hepatoprotective drugs. However, this approach suffers from numerous side effects, poor patient compliance, and unsatisfactory overall efficacy.

[0003] Traditional Chinese medicine (TCM) has advantages in preventing and treating complex metabolic diseases through multi-target and holistic regulation. Currently, some TCM compound formulas have been reported for the treatment of fatty liver or diabetes. For example, the classic formula "Huangqin Tang" (composed of Scutellaria baicalensis, Paeonia lactiflora, Glycyrrhiza uralensis, and Ziziphus jujuba) is recorded in the *Shanghan Lun* (Treatise on Cold Damage) as being used to treat damp-heat dysentery. Modern research has also confirmed its anti-inflammatory and antioxidant activities, and it is mainly used in the field of intestinal diseases.

[0004] However, there are currently no literature reports or technological insights regarding the use of Huangqin Decoction or its modified forms for the prevention and treatment of "diabetic liver disease," a complex metabolic syndrome as defined in modern medicine. More importantly, the pathogenesis of "diabetic liver disease" in Traditional Chinese Medicine often falls under the category of "liver stagnation and spleen deficiency with internal damp-heat," and frequently involves signs of deficiency and cold in the middle jiao (middle burner). Simply using cooling and heat-clearing formulas (such as the original Huangqin Decoction) may exacerbate spleen cold with long-term use, and is therefore not the optimal choice. Summary of the Invention

[0005] The purpose of this invention is to address the shortcomings of existing technologies in providing safe and effective integrated treatment solutions for "diabetic liver disease" by providing a new use of a traditional Chinese medicine composition in the prevention and treatment of this disease. This use is based on a creative improvement of a classic prescription and produces unexpected synergistic effects.

[0006] To achieve the above objectives, the following technical solution is adopted:

[0007] On the one hand, the present invention provides a traditional Chinese medicine composition for the prevention and treatment of metabolic-related fatty liver disease complicated with type 2 diabetes. The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 6-30 parts of Scutellaria baicalensis, 0-20 parts of dried ginger, 4-20 parts of Paeonia lactiflora, 4-20 parts of Glycyrrhiza uralensis, and 2-30 parts of Ziziphus jujuba.

[0008] On the other hand, the present invention also provides the application of a traditional Chinese medicine composition in the preparation of a drug for the prevention and treatment of metabolic-related fatty liver disease complicated with type 2 diabetes. The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 6-30 parts of Scutellaria baicalensis, 0-20 parts of dried ginger, 4-20 parts of Paeonia lactiflora, 4-20 parts of Glycyrrhiza uralensis, and 2-30 parts of Ziziphus jujuba.

[0009] As a preferred and inventive technical solution of the present invention, the weight part of dried ginger in the traditional Chinese medicine composition is 1 to 20 parts; the drug has good gastrointestinal tolerance with long-term use. By introducing dried ginger, this solution changes the overall formula from simply "clearing heat and drying dampness" to "harmonizing cold and warm, opening with pungent and descending with bitter", which is more in line with the core pathogenesis of "diabetic liver disease" with mixed cold and heat, and is conducive to improving the safety of long-term use.

[0010] Furthermore, the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 6-15 parts of Scutellaria baicalensis, 5-10 parts of dried ginger, 5-10 parts of Paeonia lactiflora, 5-10 parts of Glycyrrhiza uralensis, and 5-10 parts of Ziziphus jujuba.

[0011] Furthermore, the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 9 parts Scutellaria baicalensis, 6 parts dried ginger, 6 parts Paeonia lactiflora, 6 parts Glycyrrhiza uralensis, and 6 parts jujube; or, the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 9 parts Scutellaria baicalensis, 6 parts Paeonia lactiflora, 6 parts Glycyrrhiza uralensis, and 6 parts jujube.

[0012] Furthermore, when the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 9 parts Scutellaria baicalensis, 6 parts Paeonia lactiflora, 6 parts Glycyrrhiza uralensis, and 6 parts Ziziphus jujuba, the fingerprint spectrum of multiple batches of the traditional Chinese medicine composition is processed using the "Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 Edition)" to generate a fingerprint spectrum. After comparison with the retention time of a single reference standard, seven chemical components are identified: peak 1 is paeoniflorin, peak 2 is glycyrrhizin, peak 3 is baicalin, peak 4 is glycyrrhizin, peak 5 is wogonin, peak 6 is baicalein, and peak 7 is wogonin.

[0013] Furthermore, when the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 9 parts of Scutellaria baicalensis, 6 parts of dried ginger, 6 parts of Paeonia lactiflora, 6 parts of Glycyrrhiza uralensis, and 6 parts of Ziziphus jujuba, the "Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 Edition)" is used to process the chromatograms of multiple batches of the traditional Chinese medicine composition to generate fingerprint spectra. After comparing the retention time with that of a single reference standard, nine chemical components are identified, namely: peak 1 is paeoniflorin, peak 2 is glycyrrhizin, peak 3 is baicalin, peak 4 is glycyrrhizin, peak 5 is wogonin, peak 6 is baicalin, peak 7 is wogonin, peak 8 is 8-gingerol, and peak 9 is gingerone.

[0014] Furthermore, the chromatogram was detected using HPLC-DAD. The chromatographic column used was a Shiseido MGII C18 with dimensions of 4.6 mm × 250 mm and 5 μm. Gradient elution was performed using 0.1% formic acid aqueous solution (A) and acetonitrile (B) as the mobile phase. The flow rate was 1.0 mL / min, the column temperature was 35℃, and the detection wavelength was 276 nm.

[0015] Furthermore, the dosage form of the drug is an oral preparation; the oral preparation is granules, tablets, capsules, pills, or oral liquid.

[0016] Furthermore, the drug has the effect of improving insulin resistance and / or reducing fasting blood glucose; the drug has the effects of reducing hepatic lipid deposition, reducing serum transaminase levels and / or inhibiting hepatic inflammatory response.

[0017] The beneficial effects of this invention are as follows:

[0018] 1) This invention discloses the use of a specific ratio composition of Scutellaria baicalensis, Paeonia lactiflora, Glycyrrhiza uralensis, Ziziphus jujuba, and optionally dried ginger in the prevention and treatment of "diabetic liver disease", opening up new application areas for classic prescriptions;

[0019] 2) This invention creatively adds dried ginger, a warming traditional Chinese medicine, to Huangqin Decoction, forming a "combination of cold and warming" formula that can synergistically act on multiple targets of glucose and lipid metabolism, thereby producing an unexpected integrated therapeutic effect on "diabetic liver disease," resulting in a synergistic effect of "1+1>2," and significantly reducing diarrhea as an adverse reaction and improving gastrointestinal tolerance. The formula is more effective than the original formula without dried ginger in key indicators such as improving insulin sensitivity, inhibiting the expression of liver inflammatory factors, and reducing fecal water content.

[0020] 3) The drug provided by this invention can simultaneously and significantly improve abnormal glucose metabolism (reducing fasting blood glucose, improving glucose tolerance and insulin resistance) and liver metabolic damage (reducing fat deposition, reducing transaminase, and inhibiting inflammation) in "glycosuria-liver comorbidity" model animals, achieving integrated intervention on the core pathological links of the disease. Attached Figure Description

[0021] Figure 1 It is the fingerprint spectrum of traditional Chinese medicine composition 1 (excluding dried ginger); Figure 2 It is the fingerprint spectrum of traditional Chinese medicine composition 2 (containing dried ginger); Figure 3 These are the results of weight changes in each group of "diabetic liver disease" mice during the 18-week intervention period; Figure 4 The results are blood glucose test results of glucose tolerance tests in mice with comorbid diabetes and liver disease in each group after 18 weeks of intervention. Figure 5 It is the area under the OGTT curve for glucose tolerance testing in mice with comorbid diabetes after 18 weeks of intervention; Figure 6 The results are blood glucose levels from insulin sensitivity testing in mice with comorbid diabetes after 18 weeks of intervention. Figure 7 Area under the ITT curve for insulin sensitivity testing in mice with comorbid diabetes after 18 weeks of intervention; Figure 8 These are the fasting blood glucose test results of mice with comorbid diabetes and liver disease in each group after 18 weeks of intervention; Figure 9 These are the results of fasting free fatty acid detection in mice with comorbid diabetes and liver disease in each group after 18 weeks of intervention; Figure 10 These are the fasting insulin test results of mice with comorbid diabetes and liver disease in each group after 18 weeks of intervention; Figure 11 These are the results of adipose tissue-specific assessments in mice with comorbid diabetes and liver disease in each group after 18 weeks of intervention. Figure 12 The results are the assessment of systemic insulin resistance index in mice with comorbid diabetes and liver disease in each group after 18 weeks of intervention. Figure 13 The results show the deposition of triglycerides in the livers of mice with comorbid diabetes after 18 weeks of intervention. Figure 14 The results show the total cholesterol deposition in the livers of mice with comorbid diabetes after 18 weeks of intervention. Figure 15 The results show the serum triglyceride deposition in mice with comorbid diabetes after 18 weeks of intervention. Figure 16 The results show the serum total cholesterol deposition in mice with comorbid diabetes after 18 weeks of intervention. Figure 17 The results are the assessment of aspartate aminotransferase (AST) activity in each group of "glycosuria comorbidity" mice after 18 weeks of intervention; Figure 18 The results are the alanine aminotransferase (ALT) activity assessment results of mice with comorbid diabetes and liver disease in each group after 18 weeks of intervention; Figure 19 The results show the levels of interleukin-1β, a liver inflammatory factor, in mice with comorbid diabetes and liver disease in each group after 18 weeks of intervention. Figure 20 The results show the levels of interleukin-6, a liver inflammatory factor, in mice with comorbid diabetes and liver disease in each group after 18 weeks of intervention. Figure 21 These are the results of detecting the levels of tumor necrosis factor-α, an inflammatory factor in the liver of mice with comorbid diabetes, after 18 weeks of intervention. Figure 22 These are representative images of liver tissue staining in mice with comorbid diabetes after 18 weeks of intervention. Figure 23 These are the NAS scores of liver tissues from mice with comorbid diabetes after 18 weeks of intervention. Figure 24 The results are the gastrointestinal tolerance indicators of mice with comorbid diabetes and liver disease in each group after 18 weeks of intervention.

[0022] Figure 3-24 In the mean, *P<0.05, **P<0.01, and ***P<0.001. Detailed Implementation

[0034] To further illustrate the application effects and functions of the traditional Chinese medicine composition of this application, the following preferred embodiments are provided to describe in detail the specific implementation methods, technical solutions, and features according to this application. Specific features, structures, or characteristics in the various embodiments described below can be combined in any suitable form.

[0035] For experiments not specifically described in the examples, the procedures or conditions should be followed according to the conventional experimental procedures described in the literature in this field. Reagents or instruments whose manufacturers are not specified are all commercially available conventional reagent products.

[0036] On the one hand, the present invention provides a traditional Chinese medicine composition for the prevention and treatment of metabolic-related fatty liver disease complicated with type 2 diabetes. The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 6-30 parts of Scutellaria baicalensis, 0-20 parts of dried ginger, 4-20 parts of Paeonia lactiflora, 4-20 parts of Glycyrrhiza uralensis, and 2-30 parts of Ziziphus jujuba.

[0037] On the other hand, the present invention also provides the application of a traditional Chinese medicine composition in the preparation of a drug for the prevention and treatment of metabolic-related fatty liver disease complicated with type 2 diabetes. The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 6-30 parts of Scutellaria baicalensis, 0-20 parts of dried ginger, 4-20 parts of Paeonia lactiflora, 4-20 parts of Glycyrrhiza uralensis, and 2-30 parts of Ziziphus jujuba.

[0038] As a preferred and inventive technical solution of the present invention, the weight part of dried ginger in the traditional Chinese medicine composition is 1 to 20 parts. By introducing dried ginger, this solution changes the overall formula from simply "clearing heat and drying dampness" to "harmonizing cold and warm, opening with pungent and descending with bitter", which is more in line with the core pathogenesis of "diabetic liver disease" with mixed cold and heat, and is conducive to improving the safety of long-term medication.

[0039] In a preferred embodiment, the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 6-15 parts of Scutellaria baicalensis, 5-10 parts of dried ginger, 5-10 parts of Paeonia lactiflora, 5-10 parts of Glycyrrhiza uralensis, and 5-10 parts of Ziziphus jujuba.

[0040] In a preferred embodiment, the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 9 parts Scutellaria baicalensis, 6 parts dried ginger, 6 parts Paeonia lactiflora, 6 parts Glycyrrhiza uralensis, and 6 parts jujube; or, the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 9 parts Scutellaria baicalensis, 6 parts Paeonia lactiflora, 6 parts Glycyrrhiza uralensis, and 6 parts jujube.

[0041] In the embodiments, when the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 9 parts of Scutellaria baicalensis, 6 parts of Paeonia lactiflora, 6 parts of Glycyrrhiza uralensis, and 6 parts of Ziziphus jujuba, the "Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 Edition)" is used to process the chromatograms of multiple batches of the traditional Chinese medicine composition to generate fingerprint spectra. After comparison with the retention time of a single reference standard, seven chemical components are identified, namely: peak 1 is paeoniflorin, peak 2 is glycyrrhizin, peak 3 is baicalin, peak 4 is glycyrrhizin, peak 5 is wogonin, peak 6 is baicalein, and peak 7 is wogonin.

[0042] In the embodiments, the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 9 parts of Scutellaria baicalensis, 6 parts of dried ginger, 6 parts of Paeonia lactiflora, 6 parts of Glycyrrhiza uralensis, and 6 parts of Ziziphus jujuba. The "Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 Edition)" was used to process the chromatograms of multiple batches of the traditional Chinese medicine composition to generate fingerprint spectra. After comparison with the retention time of a single reference standard, nine chemical components were identified, namely: peak 1 is paeoniflorin, peak 2 is glycyrrhizin, peak 3 is baicalin, peak 4 is glycyrrhizin, peak 5 is wogonin, peak 6 is baicalin, peak 7 is wogonin, peak 8 is 8-gingerol, and peak 9 is gingerone.

[0043] In a preferred embodiment, the chromatogram is detected using HPLC-DAD, with a Shiseido MG II C18 column (4.6 mm × 250 mm, 5 μm) used as the mobile phase. Gradient elution is performed using 0.1% formic acid aqueous solution (A) and acetonitrile (B) at a flow rate of 1.0 mL / min, a column temperature of 35°C, and a detection wavelength of 276 nm.

[0044] In a preferred embodiment, the dosage form of the drug is an oral formulation.

[0045] In a preferred embodiment, the oral preparation is granules, tablets, capsules, pills, or oral liquid.

[0046] In a preferred embodiment, the drug has the effect of improving insulin resistance and / or reducing fasting blood glucose.

[0047] Preferably, the drug has the effects of reducing hepatic lipid deposition, lowering serum transaminase levels, and / or inhibiting hepatic inflammatory response; the drug has good gastrointestinal tolerance with long-term use.

[0048] Example 1: Preparation of Traditional Chinese Medicine Composition

[0049] (1) Traditional Chinese medicine composition 1 (excluding dried ginger)

[0050] Weigh out 36 g of Scutellaria baicalensis, 24 g of Paeonia lactiflora, 24 g of Glycyrrhiza uralensis, and 24 g of Ziziphus jujuba. Extract twice: first boil over high heat, then simmer over low heat. Add 10 times the amount of water to the first decoction and simmer for 60 minutes. Add 8 times the amount of water to the second decoction and simmer for 30 minutes. Filter, concentrate at 60℃~80℃, and freeze dry to obtain the freeze-dried powder of Chinese herbal composition 1.

[0051] (2) Traditional Chinese medicine composition 2 (containing dried ginger)

[0052] Weigh out 36 g of Scutellaria baicalensis, 24 g of dried ginger, 24 g of Paeonia lactiflora, 24 g of Glycyrrhiza uralensis, and 24 g of jujube. Extract twice: first boil over high heat, then simmer over low heat. Add 10 times the amount of water to the first decoction and simmer for 60 minutes. Add 8 times the amount of water to the second decoction and simmer for 30 minutes. Filter, concentrate at 60℃~80℃, and freeze dry to obtain the freeze-dried powder of Chinese herbal composition 2.

[0053] Example 2: Evaluation of the quality of traditional Chinese medicine compositions using chemical fingerprinting

[0054] 2.1 Materials: All reference standards had a purity >98%. Paeoniflorin, glycyrrhizin, baicalin, wogonin, baicalein, and wogonin were purchased from the China National Institutes for Food and Drug Control. Glycyrrhizin, 8-gingerol, and zingerone were purchased from Chengdu Manster Biotechnology Co., Ltd. All medicinal materials used in the experiments were purchased from Beijing Tongrentang Health Pharmaceutical Co., Ltd. Chromatographic grade acetonitrile was purchased from Merck AG, Germany; chromatographic grade formic acid was purchased from Anco Chemicals, USA. The experimental water was ultrapure water prepared by a pure water system (Merck Millipore, Darmstadt, Germany). All other reagents were analytical grade and purchased from Chongqing Chuandong Chemical (Group) Co., Ltd., China.

[0055] 2.2 Methods: Preparation of the test solution: Accurately weigh 0.2 g of the lyophilized powder of the traditional Chinese medicine composition (including traditional Chinese medicine composition 1 and traditional Chinese medicine composition 2, hereinafter the same), place it in an Erlenmeyer flask, accurately add 50 mL of 75% ethanol, and weigh it. Reflux at 80℃ for 30 minutes, cool, and add anhydrous ethanol to the original weight. Centrifuge the extract (8000 rpm, 5 minutes), and filter the supernatant through a 0.22 μm microporous membrane to obtain the test solution. Preparation of the reference solution: Dissolve the reference standard in a solvent to obtain the reference solution. Establishment of the chromatographic fingerprint: Chemical fingerprint analysis was performed using an Agilent 1260 Infinity high-performance liquid chromatography system (Agilent Technologies, Germany) equipped with a diode array detector. The chromatographic column was a Shiseido MGII C18 column (4.6 × 250 mm, 5 μm), and the column temperature was controlled at 35℃. The mobile phase consisted of 0.1% formic acid aqueous solution (A) and acetonitrile (B), eluted according to the following gradient program: 0 min, 17% B; 13 min, 17% B; 15 min, 20% B; 50 min, 30% B; 62 min, 40% B; 70 min, 44% B; 80 min, 90% B. The flow rate was 1.0 mL / min, the injection volume was 5 μL, and the detection wavelength was 276 nm. The chromatograms of multiple batches of samples were automatically matched and analyzed using the "Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicine" (2012 edition, Chinese Pharmacopoeia Commission).

[0056] 2.3 Results: The fingerprint spectra of traditional Chinese medicine compositions 1 and 2 are as follows: Figure 1 , 2 As shown, when the retention time of the traditional Chinese medicine composition 1 was compared with that of the single reference standard, seven chemical components were identified: peak 1 is paeoniflorin, peak 2 is glycyrrhizin, peak 3 is baicalin, peak 4 is glycyrrhizin, peak 5 is wogonin, peak 6 is baicalein, and peak 7 is wogonin.

[0057] By comparing the retention time of the traditional Chinese medicine composition 2 with that of the single reference standard, nine chemical components were identified: peak 1 is paeoniflorin, peak 2 is glycyrrhizin, peak 3 is baicalin, peak 4 is glycyrrhizin, peak 5 is wogonin, peak 6 is baicalin, peak 7 is wogonin, peak 8 is 8-gingerol, and peak 9 is gingerone.

[0058] The traditional Chinese medicine composition of the present invention should have the above-mentioned characteristic peaks. Preferably, the ratio between the integral areas of the characteristic peaks of each component should conform to the following: Figure 1 and Figure 2 The proportions of the integral areas of the characteristic peaks of each component are shown. The traditional Chinese medicine composition has... Figure 1 and Figure 2The chemical fingerprint chromatogram of traditional Chinese medicine shows the characteristic peaks and the proportional relationship of the integrated areas of each characteristic peak. The proportional relationship of the integrated areas of each characteristic peak reflects the proportional relationship of the contents of the corresponding components.

[0059] The similarity scores were all >0.9, indicating that the quality of traditional Chinese medicine compositions 1 and 2 was controllable and the consistency was good.

[0060] Example 3: Traditional Chinese medicine composition reduces weight gain in mice induced by a high-fat, high-sugar, and high-cholesterol diet.

[0061] 3.1 Experimental materials and models: C57BL / 6J mice were fed a high-fat, high-sugar, and high-cholesterol diet for 18 weeks to successfully establish an animal model of "glycosuria-hepatitis comorbidity".

[0062] 3.2 Grouping and Administration: A normal control group, a model control group, a positive drug control group (metformin, 200 mg / kg / d), a traditional Chinese medicine composition group 1 (excluding dried ginger, 1 g / kg, clinically equivalent to the human dose of 27 g / d based on body surface area), and a traditional Chinese medicine composition group 2 (containing dried ginger, 1.22 g / kg, equivalent to the amount of Scutellaria baicalensis in traditional Chinese medicine composition 1) were established. The drugs were administered via gavage concurrently with model establishment for 18 consecutive weeks.

[0063] 3.3 Detection indicators and results:

[0064] Detection indicators: such as Figure 3 As shown, compared with the model group, both the traditional Chinese medicine composition group 1 and group 2 could significantly inhibit the weight gain of mice (P < 0.001), and the weight improvement rate of the traditional Chinese medicine composition group 2 (including the dried ginger group) was better than that of the traditional Chinese medicine composition group 1 (excluding the dried ginger group).

[0065] 3.4 Conclusion: The traditional Chinese medicine composition of the present invention can effectively inhibit the excessive weight gain of mice caused by a high-fat, high-sugar, and high-cholesterol diet, and the weight improvement rate of group 2 containing the traditional Chinese medicine composition (including dried ginger group) is better than that of group 1 containing the traditional Chinese medicine composition (without dried ginger group).

[0066] Example 4: The effect of a traditional Chinese medicine composition on improving glucose tolerance and insulin resistance in mice induced by a high-fat, high-sugar, and high-cholesterol diet.

[0067] 4.1 Experimental materials and models: The experimental design is the same as in Example 3.

[0068] 4.2 Grouping and administration: Grouping is the same as in Example 3.

[0069] 4.3 Detection Indicators and Results:

[0070] (1) Detection indicators: At week 16 of intervention, mice in each group were subjected to oral glucose tolerance test (OGTT) and insulin tolerance test (ITT).

[0071] (2) Results: such as Figures 4 to 7 As shown, compared with the model group, both the traditional Chinese medicine composition groups 1 and 2 significantly reduced the area under the OGTT curve and improved glucose tolerance (P<0.001); they also significantly reduced the area under the ITT curve and improved insulin resistance (P<0.001). Both groups showed better results than the metformin-positive control group, and the improvement rates of OGTT and ITT in the traditional Chinese medicine composition group 2 (including the dried ginger group) were better than those in the traditional Chinese medicine composition group 1 (excluding the dried ginger group).

[0072] 4.4 Conclusion: The traditional Chinese medicine composition of the present invention can effectively improve glucose intolerance and insulin resistance in mice induced by a high-fat, high-sugar, and high-cholesterol diet during the intervention period, and the effect is significantly better than that of the positive control drug. Moreover, the improvement rate of OGTT and ITT in the traditional Chinese medicine composition group 2 (including the dried ginger group) is better than that in the traditional Chinese medicine composition group 1 (excluding the dried ginger group).

[0073] Example 5: The effect of a traditional Chinese medicine composition on improving systemic and adipose tissue-specific insulin resistance in mice induced by a high-fat, high-sugar, and high-cholesterol diet.

[0074] 5.1 Experimental materials and models: The experimental design is the same as in Example 3.

[0075] 5.2 Grouping and administration: Grouping was the same as in Example 3.

[0076] 5.3 Detection Indicators and Results:

[0077] (1) Fasting blood glucose detection: After the intervention, mouse serum was collected and fasting blood glucose levels were detected using a glucose kit.

[0078] (2) Fasting free fatty acid detection: Serum free fatty acid levels were measured using a free fatty acid detection kit.

[0079] (3) Fasting insulin detection: Serum insulin concentration was detected by ELISA.

[0080] (4) Insulin resistance assessment: The homeostasis model assessment of insulin resistance index (HOMA-IR) was calculated by combining fasting blood glucose and fasting insulin; the adipose tissue insulin resistance index (Adipo-IR) was calculated by combining fasting free fatty acids and fasting insulin.

[0081] (5) Results: such as Figures 8 to 10 ,as well as Figure 11 and Figure 12As shown, compared with the model group, both groups 1 and 2 of the traditional Chinese medicine composition significantly reduced fasting blood glucose, fasting free fatty acid, and fasting insulin levels (P<0.001); and significantly reduced Adipo-IR and HOMA-IR indices (P<0.001). Both groups showed better results than the metformin positive control group, and the improvement effect of group 2 of the traditional Chinese medicine composition (including dried ginger) was better than that of group 1 of the traditional Chinese medicine composition (excluding dried ginger).

[0082] 5.4 Conclusion: The traditional Chinese medicine composition of the present invention can effectively reduce fasting blood glucose, fasting free fatty acid and fasting insulin levels, and improve the specific and systemic insulin resistance of adipose tissue in mice induced by a high-fat, high-sugar and high-cholesterol diet. The effect is significantly better than that of the positive control drug. The improvement effect of the traditional Chinese medicine composition group 2 (including the dried ginger group) is better than that of the traditional Chinese medicine composition group 1 (excluding the dried ginger group).

[0083] Example 6: The effect of a traditional Chinese medicine composition on improving liver lipid deposition and elevated blood lipids in mice induced by a high-fat, high-sugar, and high-cholesterol diet.

[0084] 6.1 Experimental materials and models: The experimental design is the same as in Example 3.

[0085] 6.2 Grouping and administration: Grouping is the same as in Example 3.

[0086] 6.3 Detection Indicators and Results:

[0087] (1) Detection indicators: After the mice were euthanized, liver tissue was taken and weighed, and the content of triglycerides (TG) and total cholesterol (TC) in the liver tissue was detected; at the same time, serum was collected and the content of triglycerides (TG) and total cholesterol (TC) in the serum was detected.

[0088] (2) Results: such as Figures 13 to 16 As shown, compared with the normal control group, the liver TG ( ) of the model group mice was higher. Figure 13 ), liver TC ( Figure 14 ), serum TG ( Figure 15 ) and serum TC ( Figure 16 The content of all four indicators was significantly increased. Compared with the model group, both the traditional Chinese medicine composition group 1 and group 2 significantly reduced the above four indicators (P<0.001), and the improvement effect of traditional Chinese medicine composition group 2 (including dried ginger group) was better than that of traditional Chinese medicine composition group 1 (excluding dried ginger group).

[0089] 6.4 Conclusion: The traditional Chinese medicine composition of the present invention can effectively improve the lipid deposition in the liver of mice induced by a high-fat, high-sugar, and high-cholesterol diet (reducing liver TG and TC) and elevated blood lipids (reducing serum TG and TC). The improvement effect of the traditional Chinese medicine composition group 2 (including the dried ginger group) is better than that of the traditional Chinese medicine composition group 1 (excluding the dried ginger group).

[0090] Example 7: Protective effect of traditional Chinese medicine composition on liver damage induced by a high-fat, high-sugar, and high-cholesterol diet in mice.

[0091] 7.1 Experimental materials and models: The experimental design is the same as in Example 3.

[0092] 7.2 Grouping and administration: Grouping is the same as in Example 3.

[0093] 7.3 Detection Indicators and Results:

[0094] (1) Detection indicators: Mouse serum was collected to detect the activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT).

[0095] (2) Results: such as Figure 17 and Figure 18 As shown, compared with the normal control group, the serum AST ( ) in the model group mice was significantly higher. Figure 17 ) and ALT Figure 18 The activity was significantly enhanced. Treatment with traditional Chinese medicine compositions 1 and 2 could significantly reverse these indicators (P<0.001), indicating that they have the effect of protecting hepatocytes from damage, and the improvement rate of traditional Chinese medicine composition 2 (including dried ginger) was better than that of traditional Chinese medicine composition 1 (excluding dried ginger).

[0096] 7.4 Conclusion: The traditional Chinese medicine composition of the present invention can effectively improve liver damage in mice caused by a high-fat, high-sugar, and high-cholesterol diet, and the improvement rate of the traditional Chinese medicine composition group 2 (including the dried ginger group) is better than that of the traditional Chinese medicine composition group 1 (excluding the dried ginger group).

[0097] Example 8: The effect of a traditional Chinese medicine composition on improving liver inflammation in mice induced by a high-fat, high-sugar, and high-cholesterol diet.

[0098] 8.1 Experimental materials and models: The experimental design is the same as in Example 3.

[0099] 8.2 Grouping and administration: Grouping is the same as in Example 3.

[0100] 8.3 Detection Indicators and Results:

[0101] (1) Detection indicators: A portion of mouse liver tissue was taken to prepare homogenate, and the levels of inflammatory factors tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) were detected according to the steps of the ELISA kit.

[0102] (2) Results: The results are as follows Figures 19 to 21 As shown, compared with the normal control group, the model group mice had a higher incidence of liver inflammatory factor IL-1β (IL-1β). Figure 19 ), IL-6 ( Figure 20 ) and TNF-α ( Figure 21The levels of these indicators were significantly elevated. The traditional Chinese medicine compositions 1 and 2 significantly reversed these indicators (P<0.001), indicating that they have the effect of reducing liver inflammation. Furthermore, the improvement rate of the traditional Chinese medicine composition 2 group (containing dried ginger) was superior to that of the traditional Chinese medicine composition 1 group (excluding dried ginger).

[0103] 8.4 Conclusion: The traditional Chinese medicine composition of the present invention can effectively reduce the inflammatory response of the liver in mice caused by a high-fat, high-sugar, and high-cholesterol diet, and the improvement rate of the traditional Chinese medicine composition group 2 (including the dried ginger group) is better than that of the traditional Chinese medicine composition group 1 (excluding the dried ginger group).

[0104] Example 9: The effect of a traditional Chinese medicine composition on improving the histopathological characteristics of mouse liver tissue induced by a high-fat, high-sugar, and high-cholesterol diet.

[0105] 9.1 Experimental materials and models: The experimental design is the same as in Example 3.

[0106] 9.2 Grouping and administration: Grouping is the same as in Example 3.

[0107] 9.3 Detection Indicators and Results:

[0108] (1) Detection indicators: Mouse liver tissue was taken, some of which were fixed with 4% paraformaldehyde, embedded in paraffin and sectioned. Hematoxylin-eosin (H&E), Masson staining and Oil Red O (ORO) staining were performed and observed under an optical microscope. The non-alcoholic fatty liver disease activity score (NAS) scoring system was used to semi-quantitatively assess the pathological changes in the liver.

[0109] (2) Results: such as Figure 22 and Figure 23 As shown, H&E staining of liver tissue from the model group mice revealed significant hepatocyte ballooning degeneration and inflammatory cell infiltration, as well as Masson staining and ORO staining. Figure 22 The results indicated collagen fiber deposition and lipid accumulation, respectively; and the pathological changes in groups 1 and 2 of the traditional Chinese medicine composition were significantly alleviated. Furthermore, compared with the model group, both groups 1 and 2 of the traditional Chinese medicine composition significantly reduced the NAS score. Figure 23 (P<0.001) The improvement effect of the Chinese medicine composition group 2 (including the dried ginger group) was better than that of the Chinese medicine composition group 1 (excluding the dried ginger group).

[0110] 9.4 Conclusion: This invention has demonstrated the protective effect of the traditional Chinese medicine composition on the liver at the histological level. It can effectively reduce hepatocyte ballooning degeneration, inflammatory cell infiltration, collagen fiber deposition and lipid accumulation, and reduce NAS score. Moreover, the improvement effect of the traditional Chinese medicine composition group 2 (including the dried ginger group) is better than that of the traditional Chinese medicine composition group 1 (excluding the dried ginger group).

[0111] Example 10: Effects of Traditional Chinese Medicine Composition on Gastrointestinal Tolerance in Mice

[0112] 10.1 Experimental materials and models: The experimental design is the same as in Example 3.

[0113] 10.2 Grouping and administration: Grouping is the same as in Example 3.

[0114] 10.3 Detection Indicators and Results:

[0115] (1) Detection indicators: At week 17 of the intervention, fresh feces were collected into sample tubes, weighed, dried at 105℃ for 24 h until constant weight, and weighed again to calculate the fecal moisture content. Fecal moisture content % = [(wet weight - dry weight) ÷ wet weight] × 100%.

[0116] (2) Results: The results are as follows Figure 24 As shown, compared with the normal control group, the fecal water content of mice in the model group did not change significantly. Compared with the model group, the fecal water content of mice in the positive drug control group and the traditional Chinese medicine composition 1 group (excluding dried ginger group) increased significantly, while the fecal water content of mice in the traditional Chinese medicine composition 2 group (including dried ginger group) did not differ significantly.

[0117] 10.4 Conclusion: This invention confirms that the two groups of traditional Chinese medicine compositions (including the dried ginger group) have good gastrointestinal tolerance after long-term use.

[0118] The above description is merely a preferred embodiment of this application; however, the scope of protection of this application is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in this application, based on the technical solution and its improved concept, should be covered within the scope of protection of this application.

Claims

1. A traditional Chinese medicine composition for the prevention and treatment of metabolic-related fatty liver disease complicated with type 2 diabetes, characterized in that, The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 6-30 parts of Scutellaria baicalensis, 0-20 parts of dried ginger, 4-20 parts of Paeonia lactiflora, 4-20 parts of Glycyrrhiza uralensis, and 2-30 parts of jujube.

2. The application of a traditional Chinese medicine composition in the preparation of a drug for the prevention and treatment of metabolic-related fatty liver disease complicated with type 2 diabetes, characterized in that, The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 6-30 parts of Scutellaria baicalensis, 0-20 parts of dried ginger, 4-20 parts of Paeonia lactiflora, 4-20 parts of Glycyrrhiza uralensis, and 2-30 parts of jujube.

3. The application according to claim 2, characterized in that, The traditional Chinese medicine composition contains 1 to 20 parts by weight of dried ginger; the drug has good gastrointestinal tolerance with long-term use.

4. The application according to claim 2, characterized in that, The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 6-15 parts of Scutellaria baicalensis, 5-10 parts of dried ginger, 5-10 parts of Paeonia lactiflora, 5-10 parts of Glycyrrhiza uralensis, and 5-10 parts of Ziziphus jujuba.

5. The application according to claim 2, characterized in that, The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 9 parts Scutellaria baicalensis, 6 parts dried ginger, 6 parts Paeonia lactiflora, 6 parts Glycyrrhiza uralensis, and 6 parts jujube; or, the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 9 parts Scutellaria baicalensis, 6 parts Paeonia lactiflora, 6 parts Glycyrrhiza uralensis, and 6 parts jujube.

6. The application according to claim 5, characterized in that, When the weight of dried ginger in the traditional Chinese medicine composition is 0 parts, the "Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 Edition)" is used to process the chromatograms of multiple batches of the traditional Chinese medicine composition to generate fingerprint spectra. By comparing the retention time with that of a single reference standard, seven chemical components are identified: peak 1 is paeoniflorin, peak 2 is glycyrrhizin, peak 3 is baicalin, peak 4 is glycyrrhizin, peak 5 is wogonin, peak 6 is baicalin, and peak 7 is wogonin.

7. The application according to claim 5, characterized in that, When the weight of dried ginger in the traditional Chinese medicine composition is 6 parts, the "Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 Edition)" is used to process the chromatograms of multiple batches of the traditional Chinese medicine composition to generate fingerprint spectra. After comparison with the retention time of a single reference standard, 9 chemical components are identified, namely: peak 1 is paeoniflorin, peak 2 is glycyrrhizin, peak 3 is baicalin, peak 4 is glycyrrhizin, peak 5 is wogonin, peak 6 is baicalin, peak 7 is wogonin, peak 8 is 8-gingerol, and peak 9 is gingerone.

8. The application according to claim 6 or 7, characterized in that, The chromatograms were detected using HPLC-DAD. The chromatographic column used was a Shiseido MG II C18 with dimensions of 4.6 mm × 250 mm and 5 μm. Gradient elution was performed using 0.1% formic acid aqueous solution (A) and acetonitrile (B) as the mobile phase. The flow rate was 1.0 mL / min, the column temperature was 35℃, and the detection wavelength was 276 nm.

9. The application according to claim 2, characterized in that, The dosage form of the drug is an oral preparation; the oral preparation is granules, tablets, capsules, pills or oral liquid.

10. The application according to claim 2, characterized in that, The drug has the effect of improving insulin resistance and / or reducing fasting blood glucose; the drug has the effect of reducing hepatic lipid deposition, reducing serum transaminase levels and / or inhibiting hepatic inflammatory response.

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