Application of Wuminan pharmaceutical composition or preparation of Wuminan pharmaceutical composition in preparation of medicine for preventing or treating non-alcoholic steatohepatitis

Abstract

The invention relates to the field of medical application, in particular to application of a Wumingdan pharmaceutical composition or a preparation of the Wumingdan pharmaceutical composition in preparation of medicine for preventing or treating non-alcoholic steatohepatitis, and the Wumingdan pharmaceutical composition comprises schisandra chinensis, antelope horn and salvia miltiorrhiza. The Wumingdan pharmaceutical composition or the preparation (such as Wumingdan capsules) of the Wumingdan pharmaceutical composition is applied to research on treatment of non-alcoholic steatohepatitis for the first time, and new application of a traditional compound traditional Chinese medicine is developed from the angle of new application of old medicine. A non-alcoholic steatohepatitis mouse model is copied by feeding low methionine and choline-deficient high fat mouse food (CDAHFD), and the influence of the Wuminan capsule on serum biochemical indexes, liver tissue fat content and liver tissue pathological changes of the model mouse is observed, so that the drug has a treatment effect on the non-alcoholic steatohepatitis.

Description

Technical Field

[0001] The present invention relates to the field of medical applications, and in particular to an application of a Wulingdan pharmaceutical composition or a preparation thereof in preparing a drug for preventing or treating nonalcoholic fatty hepatitis. Background Art

[0002] In recent years, the disease spectrum of chronic liver diseases in my country has changed significantly. With the aging of the population, the effective prevention and treatment of viral hepatitis, and the prevalence of obesity and alcohol abuse, fatty liver disease (FLD) is becoming a new major health problem in my country. Among them, non-alcoholic fatty liver disease (NAFLD) is a series of liver diseases closely related to obesity, type 2 diabetes and metabolic syndrome. It is defined as a disease in which more than 5% of liver cells have fatty degeneration in the absence of almost no alcohol intake, viral infection or other specific factors. With the significant increase in the incidence of obesity and type 2 diabetes, NAFLD has become one of the most common chronic liver diseases worldwide, with a prevalence of about 25%. NAFLD can affect people of all ages from infants to the elderly. According to a sample survey of the general population in large and medium-sized cities, the prevalence of NAFLD in adults in my country is about 15%. In patients with chronic hepatitis of unknown etiology, liver biopsy histological examination found that the prevalence of non-alcoholic fatty liver disease (NASH) was 16%; in obese patients with abnormal liver enzymes, the prevalence of NASH was 34%, and NASH patients have a great risk of progressing to end-stage liver disease (liver fibrosis, cirrhosis and hepatocellular carcinoma). Regarding the pathogenesis of NASH, the "double-hit" theory is generally accepted. As the research deepens, some scholars have proposed the "multiple-hit" theory. In short, NASH is a common clinical disease with complex pathogenic factors. Western medicine treatment is mainly aimed at the pathogenic risk factors of NASH and the use of liver-protecting drugs, but there is no specific means. Therefore, exploring a new treatment method is of great clinical significance.

[0003] Traditional Chinese medicine has its own unique theory and rich clinical experience in treating NASH. There is no name for non-alcoholic fatty hepatitis in the literature of past dynasties. According to its clinical symptoms, it can be attributed to the categories of "flank pain", "fullness", "fat gas", "liver habit", "accumulation", etc. NASH is not recorded in ancient relevant documents, but there are similar records. The earliest related record can be found in "Nanjing·Fifty-six Difficulties": "The accumulation of the liver is called fat gas", suggesting that the occurrence of this disease is related to the love of eating fatty and sweet food. Traditional Chinese medicine etiology believes that the occurrence of obesity and its complications is related to congenital endowment, overeating fatty and sweet food, neglect of work, excessive seven emotions, spleen and stomach deficiency, phlegm and water dampness, etc. Among them, the relationship with phlegm and water is the closest. "Phlegm" refers to the metabolic products or pathological products accumulated in the body during the physiological process or pathological changes of the human body. From the perspective of modern doctors' understanding of this disease, the main location of this disease is in the liver, spleen and stomach, involving basic pathological factors such as phlegm and dampness, qi stagnation, and heat pathogen plays an important role in the development of this disease. The disease is not cured for a long time, and it is prolonged for a long time, blood stasis is endogenous, and multiple pathological factors interact and influence each other, causing the disease to gradually develop. Wulingdan Capsule is composed of Schisandra chinensis, Antelope Horn, and Salvia miltiorrhiza. It is summarized by Professor Guan Youbo, a national famous old Chinese medicine doctor, based on the etiology and pathogenesis of viral hepatitis and clinical characteristics, after many years of clinical practice. It has the effects of invigorating qi and activating blood circulation, cooling the liver and detoxifying, and is used to treat chronic hepatitis B with a certain effect. On this basis, the inventor further studied this composition and unexpectedly found that this composition has an obvious effect in treating non-alcoholic fatty hepatitis, which is expected to fill the defects of drugs for treating non-alcoholic fatty hepatitis. Summary of the invention

[0004] Therefore, the object of the present invention is to provide a new use of a Wulingdan pharmaceutical composition or a preparation thereof, namely, the use of a Wulingdan pharmaceutical composition or a preparation thereof in the preparation of a medicament for preventing or treating non-alcoholic fatty hepatitis.

[0005] To this end, the present application provides the use of a Wulingdan pharmaceutical composition or a preparation thereof in the preparation of a medicament for preventing or treating non-alcoholic fatty hepatitis.

[0006] Furthermore, the preparation uses the Wulingdan pharmaceutical composition as an active ingredient.

[0007] Furthermore, the preparation also includes pharmaceutically acceptable excipients.

[0008] The Wulingdan pharmaceutical composition or its preparation can be prepared in-house according to conventional methods in the art, or purchased from the pharmaceutical factory of Beijing Tongrentang Technology Development Co., Ltd.

[0009] Furthermore, the preparation also includes pharmaceutically acceptable excipients; preferably, the pharmaceutically acceptable excipients include one or more of fillers, disintegrants, lubricants, suspending agents, binders, sweeteners, flavoring agents, and preservatives.

[0010] Furthermore, the preparation includes a liquid preparation or a solid preparation.

[0011] Furthermore, the solid preparation includes powders, tablets, capsules, granules or pills; and / or the liquid preparation includes injections, mixtures, syrups or suspensions.

[0012] Furthermore, the drug has a protective effect on liver damage in mice with non-alcoholic fatty liver disease and can alleviate hepatic inflammatory cell infiltration.

[0013] Furthermore, the drug can reduce the levels of ALT and AST in the serum of mice with non-alcoholic fatty liver disease. ALT is alanine aminotransferase, and AST is aspartate aminotransferase. ALT and AST are the most widely used biochemical indicators reflecting liver cell damage in clinical practice.

[0014] Furthermore, the drug can reduce the TC and TG contents in the liver tissue of mice with non-alcoholic fatty liver disease. TC is total cholesterol, and TG is triglyceride.

[0015] Furthermore, the non-alcoholic fatty liver disease mouse model is caused by a low methionine-choline deficient diet.

[0016] The technical solution of the present invention has the following advantages:

[0017] The present invention is the first to use Wulingdan pharmaceutical composition or its preparation (such as Wulingdan capsule) for the study of treating non-alcoholic fatty hepatitis, and develops new uses of traditional compound Chinese medicine from the perspective of "new uses of old medicine". The present invention replicates the non-alcoholic fatty hepatitis mouse model by feeding low methionine and choline-deficient high-fat mouse diet (CDAHFD), observes the effects of Wulingdan capsule on serum biochemical indicators, liver tissue fat content and liver tissue pathological changes of model mice, and finds that the drug has a therapeutic effect on non-alcoholic fatty hepatitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0019] Figure 1 The effect of Wulingdan capsule on the pathological changes of liver tissue in mice with non-alcoholic fatty liver disease model. A: blank group; B: model group; C: silymarin capsule group; D: Wulingdan capsule high-dose group; E: Wulingdan capsule medium-dose group; F: Wulingdan capsule low-dose group. DETAILED DESCRIPTION

[0020] The following examples are provided for a better understanding of the present invention, but are not intended to limit the best mode of implementation, nor to limit the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior arts shall fall within the protection scope of the present invention.

[0021] If no specific experimental steps or conditions are specified in the examples, the conventional experimental steps or conditions described in the literature in the field can be used. If no manufacturer is specified for the reagents or instruments used, they are all conventional reagent products that can be obtained commercially.

[0022] The Wulingdan capsules (National Medicine Standard Z11020305) used in the study were provided by the Beijing Tongrentang Technology Development Co., Ltd. Pharmaceutical Factory. Specification: 0.3 g per capsule. Batch number: 18160935. Before use, the drug powder of the Wulingdan capsules was removed from the shell and dissolved in water to prepare concentrations of 27, 13.5, and 6.75 mg mL -1 of drug solution.

[0023] Silymarin capsules were purchased from MADAUS GMBH. Specification: 140 mg*10 capsules. Batch number: B1903201. Before use, remove the shell of silymarin capsules and dissolve the powder in water to obtain a concentration of 3.15 mg·mL -1 of drug solution.

[0024] Experimental Example 1

[0025] 1. Model preparation and drug administration method

[0026] C57BL / 6J mice, male, were randomly divided into 6 groups according to body weight after adaptive feeding, namely blank group, model group, silymarin capsule group, Wulingdan capsule high, medium and low dose groups, with 12 mice in each group. Except for the blank group, the mice in the other groups were given low methionine and choline-deficient high-fat mouse diet (purchased from Deci Biotechnology (Wuxi) Co., Ltd., batch number: 20210305) daily, free to eat, and the non-alcoholic fatty liver disease mouse model was replicated for 4 weeks. The blank group mice were given ordinary maintenance feed and drinking water for mice. While the model was being replicated, each group was given the corresponding dose of drugs by gavage (see Table 1 for specific dosages), once a day, for 4 consecutive weeks.

[0027] Table 1 Dosage and concentration of mice in each group

[0028]

[0029]

[0030] 2. Detection method

[0031] (1) Determination of serum ALT and AST: After the last administration, the subjects were anesthetized by intraperitoneal injection of 60 mg / kg·BW sodium pentobarbital solution, and blood was collected from the inferior vena cava. The blood was centrifuged at 3500 r / min for 10 min, and the serum was stored in a -80°C refrigerator. The levels of ALT and AST in serum were detected according to the instructions of the kit (full name of the kit: ALT kit (enzyme reader method), manufacturer: Nanjing Jiancheng Bioengineering Institute, batch number: 20210507; full name of the kit: AST kit (enzyme reader method), manufacturer: Nanjing Jiancheng Bioengineering Institute, batch number: 20210506).

[0032] (2) Histopathological examination of the liver: The left lobe of the liver was placed in paraformaldehyde fixative for 48 h. A cross-section was made from the middle of the left lobe of the liver. After dehydration, transparency, wax immersion, embedding, sectioning, dewaxing, and HE staining, the morphological changes of the liver tissue were observed under an optical microscope.

[0033] (3) Determination of TC and TG in liver tissue homogenate: The remaining liver tissue was quickly frozen in liquid nitrogen and transferred to a -80°C refrigerator for freezing after the dissection was completed. 10% tissue homogenate was prepared under low temperature conditions of liver tissue, and the contents of various indicators in the liver tissue homogenate were detected using an ELISA instrument according to the instructions of the kit (full name of the kit: Total Cholesterol (T-CHO) Test Kit, manufacturer: Nanjing Jiancheng Bioengineering Institute, batch number: 20210508; full name of the kit: Triglyceride (TG) Test Kit, manufacturer: Nanjing Jiancheng Bioengineering Institute, batch number: 20210916).

[0034] 3. Test results

[0035] (1) Effects on serum ALT and AST levels in mice

[0036] The experimental results showed that the ALT and AST levels in the serum of the mice in the model group were significantly increased, which was significantly different from the blank group (p<0.01), indicating that the liver tissue of the model mice was significantly damaged. The serum ALT and AST levels of the mice in the silymarin capsule group were significantly reduced, which was significantly different from the model group (p<0.01). The serum ALT level of the mice in the high-dose Wulingdan capsule group was significantly lower than that in the model group, and the difference was significant (p<0.05); the serum ALT and AST levels of the mice in the medium and low-dose Wulingdan capsule groups were significantly reduced, which was significantly different from the model group (p<0.05, p<0.01, respectively). The results showed that Wulingdan capsule had a significant improvement on the liver function of mice with non-alcoholic fatty liver disease (see Table 2 for experimental results).

[0037] Table 2 Effects of Wulingdan Capsule on serum biochemical indexes in mice with nonalcoholic fatty liver disease

[0038]

[0039]

[0040] Compared with the model group: * p<0.05; ** p<0.01.

[0041] (2) Effects on TC and TG contents in mouse liver tissue

[0042] The experimental results showed that the TG content in the liver tissue of the model group mice was significantly increased, and there was a significant difference compared with the blank group (p<0.01), indicating that the fat content in the liver tissue of the model mice increased, forming a fatty liver model. The TG content in the liver tissue of the mice in the high- and low-dose Wulingdan capsule groups was significantly lower than that in the model group, and the difference was significant (p<0.05), indicating that Wulingdan capsule had a significant effect on reducing the fat content in the liver tissue of the model mice (see Table 3 for experimental results).

[0043] Table 3 Effects of Wulingdan Capsule on TC and TG contents in liver tissue of mice with nonalcoholic fatty liver disease

[0044]

[0045]

[0046] Compared with the model group: * p<0.05; ** p<0.01.

[0047] (3) Effects on pathological changes of mouse liver tissue

[0048] The experimental results showed that the model mice had liver tissue damage, unclear liver lobule structure, narrow liver sinusoids, obvious vacuoles in a large number of hepatocytes, hepatocyte nuclei squeezed to one side, some hepatocytes dissolved, interstitial inflammatory cells infiltrated, and cell necrosis. According to the NAS scoring standard, the total NAS score of the model group was significantly different from that of the blank group (p<0.01). All dose groups of Wulingdan Capsule can reduce the NAS score of liver tissue of model mice, which is significantly different from that of the model group (p<0.05), and have a good alleviating effect on liver inflammatory cell infiltration. It is suggested that Wulingdan Capsule has a good protective effect on liver damage in mice with non-alcoholic fatty liver disease caused by this experimental condition, which is mainly manifested in the inhibitory effect on inflammatory cell infiltration (see Table 4 for experimental results).

[0049] Table 4 Effect of Wulingdan Capsule on NAS score of liver histopathology in mice with nonalcoholic fatty liver disease

[0050]

[0051]

[0052]

[0053] Compared with the model group: * p<0.05; ** p<0.01.

[0054] In summary, this invention is the first to use Wulingdan Capsules for the prevention and treatment of non-alcoholic fatty liver disease, and develops new uses for traditional compound Chinese medicine from the perspective of "new uses for old medicines". By conducting a pharmacodynamic study on the treatment of non-alcoholic fatty liver disease with Wulingdan Capsules, the therapeutic effect of the drug on the disease is clarified, the new uses of the drug are expanded, and a new therapeutic drug and means are provided for the clinical treatment of NASH, thereby providing technical support for clinical precision drug use.

[0055] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. Use of the Wulingdan pharmaceutical composition or its preparation in the preparation of a medicament for preventing or treating non-alcoholic fatty hepatitis.

2. The use according to claim 1, characterized in that The preparation uses the Wulingdan pharmaceutical composition as an active component.

3. The use according to claim 1 or 2, characterized in that The Wulingdan medicinal composition includes Schisandra chinensis, antelope horn and Salvia miltiorrhiza.

4. The use according to any one of claims 1 to 3, characterized in that: The preparation further includes pharmaceutically acceptable excipients; preferably, the pharmaceutically acceptable excipients include one or more of fillers, disintegrants, lubricants, suspending agents, binders, sweeteners, flavoring agents, and preservatives.

5. The use according to any one of claims 1 to 4, characterized in that: The preparation includes a liquid preparation or a solid preparation.

6. The use according to claim 5, characterized in that The solid preparation includes powder, tablet, capsule, granule or pill; and / or the liquid preparation includes injection, mixture, syrup or suspension.

7. The use according to any one of claims 1 to 6, characterized in that: The drug has a protective effect on liver damage in mice with non-alcoholic fatty liver disease and can alleviate hepatic inflammatory cell infiltration.

8. The use according to any one of claims 1 to 7, characterized in that: The drug can reduce the levels of ALT and AST in the serum of mice with non-alcoholic fatty liver disease.

9. The use according to any one of claims 1 to 8, characterized in that The drug can reduce the TC and TG contents in liver tissues of mice with non-alcoholic fatty liver disease.

10. The use according to any one of claims 1 to 9, characterized in that: The nonalcoholic steatohepatitis mouse model is induced by a low methionine-choline deficient diet.

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