Application of apolipoprotein H in preparation of drugs or health care products for preventing and / or treating alcoholic diseases

Abstract

The invention discloses application of apolipoprotein H in preparation of drugs or health care products for preventing and / or treating alcoholic diseases. Researches show that an alcoholic hepatitis model is constructed by taking 10-week-old ApoH gene knockout mice (C57BL / 6ApoH- / -) as a background, a wild C57BL / 6 mouse alcoholic hepatitis model is taken as a contrast, peripheral blood transaminase and blood fat level, liver tissue fat content and liver tissue immunohistochemical staining are detected, and a result shows that the TC content of the liver tissue of the ApoH- / -mouse is obviously increased. The composition and content of bile acid in liver tissues and intestinal contents and the composition and abundance of intestinal flora are further detected, and results indicate that metabolic disorder of bile acid and disorder of intestinal flora exist. Based on the mechanism that low expression of alcoholic apolipoprotein H causes disorder of intestinal flora, affects bile acid metabolism and aggravates hepatic cell adiposis, the application is an accurate targeted therapeutic drug for treating alcoholic multi-organ injury.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and particularly to the application of apolipoprotein H in the preparation of medicaments or health products for preventing and / or treating alcoholic diseases. Background Art

[0002] In recent years, there have been an increasing number of patients with alcoholic diseases clinically. From a medical classification perspective, alcoholic diseases can be divided into acute alcoholic diseases and chronic alcoholic diseases. Acute alcoholic diseases are neurological problems and intestinal disorders that occur after a large amount of alcohol consumption at one time. Chronic alcoholic diseases mostly occur in patients with chronic alcohol dependence. Due to long-term alcohol dependence, there will be weakness in the limbs, often accompanied by peripheral nerve problems. At the same time, it affects the digestive system, circulatory system, and endocrine system, and complications such as alcoholic dementia, chronic gastritis, alcoholic fatty liver, coronary heart disease, and pancreatitis occur, especially gastrointestinal diseases and liver diseases are more common.

[0003] Intestinal flora dysbiosis is a relatively common gastrointestinal disease. Mainly under certain pathological conditions of the intestinal flora in the human body, the destruction of the intestinal barrier can lead to the translocation of bacteria and their metabolites and the abnormal activation of the immune system, causing damage to the intestine and liver.

[0004] Therefore, it is particularly important to develop a medicament or health product that can prevent and / or treat alcoholic diseases. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide the application of apolipoprotein H in the preparation of medicaments or health products for preventing and / or treating alcoholic diseases, which solves the technical problem of multi-organ damage caused by alcohol.

[0006] The technical problem to be solved by the present invention is achieved through the following technical solutions:

[0007] An object of the present application is to provide the application of apolipoprotein H in the preparation of medicaments or health products for preventing and / or treating alcoholic diseases.

[0008] Preferably, the medicament or health product contains apolipoprotein H or the apolipoprotein H gene, and the medicament or health product can maintain the homeostasis of the intestinal microecology and the balance of bile acid metabolism.

[0009] Preferably, the medicament or health product is used to alleviate the abnormal bile acid metabolism caused by alcoholic injury, and to alleviate the intestinal microecological disorder secondary to alcoholic injury.

[0010] Preferably, the alcoholic disease is alcoholic gastrointestinal injury and / or abnormal or imbalanced bile acid metabolism, the alcoholic intestinal injury is intestinal microecological disorder, and the abnormal or imbalanced bile acid metabolism is bile acid metabolic disorder.

[0011] Preferably, the drug or health product further comprises a pharmaceutically acceptable excipient, and the drug is an injection, tablet, granule or oral preparation, or a genetically engineered drug.

[0012] The second object of the present application is to provide an application of apolipoprotein H as a target in the preparation of a drug for preventing and / or treating alcoholic diseases, and the drug can maintain the homeostasis of intestinal microecology and the balance of bile acid metabolism.

[0013] Preferably, the alcoholic disease is alcoholic gastrointestinal injury and / or abnormal or imbalanced bile acid metabolism, the alcoholic intestinal injury is intestinal microecological disorder, and the abnormal or imbalanced bile acid metabolism is bile acid metabolic disorder.

[0014] The third object of the present application is to provide an application of a reagent for detecting a biomarker in the preparation of a product for diagnosing alcoholic diseases, and the biomarker is apolipoprotein H.

[0015] Preferably, the reagent for detecting the biomarker is a reagent for detecting the expression level of apolipoprotein H.

[0016] The above technical solution of the present invention has the following beneficial effects:

[0017] Based on the mechanism that low expression of alcoholic apolipoprotein H leads to intestinal microecological disorder, affects bile acid metabolism, and simultaneously aggravates hepatocyte steatosis, the present application provides a precision targeted therapeutic drug for treating alcoholic multi-organ injury. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The drawings incorporated in the specification and constituting a part of the specification illustrate embodiments of the present invention and, together with the description, are used to explain the principles of the present invention.

[0019] Figure 1 Results of liver transcriptome detection and analysis for patients with alcoholic liver disease (wherein: a is differential gene analysis of liver transcriptome, b is change in liver APOH gene expression, c is enrichment analysis of alcoholic hepatitis and bile acid metabolism pathway, d is enrichment analysis of apolipoprotein H expression change and bile acid metabolism pathway).

[0020] Figure 2 Results of liver transcriptome detection and analysis for an alcoholic fatty liver model of gene knockout mice (wherein: a is mouse ApoH gene expression, b is enrichment analysis of cholesterol metabolism pathway in the liver of model mice, c is genes related to cholesterol metabolism).

[0021] Figure 3 Results of bile acid detection and analysis in the liver and intestine (where: a is the bile acid content in the liver, and b is the bile acid content in the intestinal contents).

[0022] Figure 4 Results of intestinal flora detection and its correlation analysis with bile acids (where: a is the change in intestinal flora abundance, b is the bile acid metabolism-related flora, c is the flora function enrichment analysis, d is the correlation analysis between intestinal flora and intestinal bile acid content, and e is the correlation analysis between intestinal flora and liver bile acid). Detailed implementation manners

[0023] Various exemplary embodiments of the present invention will now be described in detail with reference to the accompanying drawings. It should be noted that: Unless otherwise specifically stated, the relative arrangements of components and steps, numerical expressions, and numerical values set forth in these embodiments do not limit the scope of the present invention.

[0024] In the following embodiments of the present application, the products and technologies used, unless otherwise specified, are all regarded as adopting conventional operations.

[0025] Alcohol intake directly leads to intestinal microecological disorders. Currently, it is considered that the intestinal microecology and bile acid metabolism have an interactive effect. Abnormal bile acid metabolism is one of the main regulatory factors in alcoholic diseases, and the gut-liver axis plays an important regulatory role in the pathogenesis of alcoholic diseases.

[0026] Studies have shown that using 10-week-old ApoH gene knockout mice (C57BL / 6ApoH - / - ) as the background to construct an alcoholic hepatitis model (chronic-plus-binge model), using the alcoholic hepatitis model of wild-type C57BL / 6 mice as a control, detecting the levels of peripheral blood transaminases and blood lipids, the fat content in liver tissue, and immunohistochemical staining of liver tissue. The results show that the TC content in the liver tissue of ApoH - / - mice is significantly increased. Further detecting the composition and content of bile acids in liver tissue and intestinal contents, as well as the composition and abundance of intestinal flora, the results suggest abnormal bile acid metabolism and intestinal microecological disorders. Based on the mechanism that low expression of alcoholic apolipoprotein H leads to intestinal microecological disorders, affects bile acid metabolism, and at the same time exacerbates hepatocyte steatosis, the present application provides a precision-targeted therapeutic drug for treating alcoholic multi-organ damage.

[0027] Example 1 Analysis of apolipoprotein H and alcoholic diseases

[0028] Figure 1 Results of liver transcriptome detection and analysis in the alcoholic liver population. As Figure 1 shown, we found that the expression of the apolipoprotein H gene in patients with alcoholic hepatitis was significantly decreased, and there was a significant difference compared with the healthy control group ( Figure 1Figures a and b). There are abnormalities in bile acid metabolism in patients with alcoholic hepatitis ( Figure 1 Figure c), and further analysis found that the change in apolipoprotein H expression is closely related to bile acid metabolic activity. In those with relatively high expression, the bile acid synthesis ability is enhanced; in those with relatively low expression, the bile acid transport and secretion abilities are significantly reduced ( Figure 1 Figure d). The regulatory mechanism of apolipoprotein H and alcoholic diseases in the gut-liver axis direction has not been elucidated, so a mouse model was constructed for in-depth research.

[0029] Gene knockout mice were constructed. This method can refer to the existing technology and will not be elaborated here. An alcoholic hepatitis model (chronic-plus-binge ethanol feeding mouse model) was constructed under the background of APOH gene knockout mice. Non-alcohol-fed gene knockout mice were used as controls, and the model phenotypes and changes were identified by detecting serum transaminases, blood lipids, liver tissue fat content, and liver tissue pathological staining.

[0030] The results showed that: the cholesterol content in the liver tissue of alcohol-fed gene knockout mice increased, and steatosis worsened. Further, liver transcriptome detection was performed to analyze differential gene expression and functional enrichment. The results suggested that: the differential genes of alcohol-fed gene knockout mice were enriched in the cholesterol metabolism pathway, including bile acid synthesis and secretion (such as Figure 2 shown). Therefore, we speculated that alcohol-fed gene knockout mice are involved in the regulation of bile acid metabolism.

[0031] Furthermore, the bile acid content in the liver tissue and intestinal contents of the mice was continuously detected. Analysis found that there were obvious differences in the bile acid composition and content in the liver tissue and intestinal contents (such as Figure 3 shown). The composition ratios of taurocholic acid (TCA) and hyocholic acid (HCA) in the liver tissue of alcohol-fed gene knockout mice were significantly increased, and the content of taurine-conjugated secondary bile acids was significantly increased. The contents of cholic acid (CA) and hyocholic acid (HCA) were also significantly increased. In addition, in the intestinal contents, the composition ratios and contents of TUDCA and βUDCA were significantly decreased. Therefore, we believe that not only are there changes in the synthesis, transport, and secretion of bile acids, but the enterohepatic circulation of bile acids is also affected.

[0032] Furthermore, the changes in the intestinal flora of the model mice were detected. Through diversity and abundance analysis, as well as differential flora function prediction, it was found that: there were obvious differences in the intestinal flora abundance and composition of alcohol-fed gene knockout mice ( Figure 4 Figures a and b), and functional prediction analysis suggested that the differential flora was related to metabolic functions ( Figure 4 Figure c), and then correlation analysis was performed with the bile acid content ( Figure 4(in Figure d), the gut microbiota associated with the changes in bile acid levels was identified.

[0033] In summary, low expression of alcoholic APOH can directly lead to gut microbiota dysbiosis, affect bile acid metabolism, and further exacerbate hepatic steatosis. Abnormal bile acid metabolism is one of the main regulatory factors in alcoholic multi-organ injury.

[0034] Example 2: High expression of alcoholic APOH

[0035] 1. Under the background of the above-mentioned alcohol-fed mouse model, recombinant APOH protein was exogenously supplemented through the tail vein of mice. The non-injected group was used as a control. Peripheral blood, intestinal tissues, and intestinal contents of mice were collected at different time points after injection to examine the composition and abundance changes of the gut microbiota in model mice, the functional enrichment of differential microbiota, the changes in bile acid metabolism, the correlation analysis between bile acid changes and gut microbiota, and the pathological staining of liver and intestinal tissues to identify the degree of organ injury.

[0036] 2. Targeting the mouse liver to overexpress the ApoH gene, an alcohol-fed model was further constructed. Peripheral blood, liver and intestinal tissues, and intestinal contents of mice were collected at different times to examine the composition and abundance changes of the gut microbiota in model mice, the functional enrichment of differential microbiota, the changes in bile acid metabolism, the correlation analysis between bile acid changes and gut microbiota, and the pathological staining of liver and intestinal tissues to identify the degree of organ injury.

[0037] 3. Under the background of the above-mentioned alcohol-fed mouse model, antibiotics were orally administered to construct germ-free mice. The group receiving antibiotics was used as a control. Peripheral blood and liver tissues of mice were collected at different time points to detect the expression changes of APOH gene and protein, so as to further clarify that the expression of APOH is directly related to the changes in gut microbiota.

[0038] The results showed that high expression of APOH can restore the homeostasis of alcohol-induced gut microbiota dysbiosis and reduce the degree of abnormal bile acid metabolism. Further suggesting that detecting the APOH level and increasing the APOH content in the body to maintain it normal is of great significance for the prevention and treatment of gut microbiota dysbiosis and multi-organ injury caused by alcohol.

[0039] Through the study of apolipoprotein H, it was found that it can be used as a detection target for alcoholic diseases, making the diagnosis and treatment of alcoholic diseases easier. Treatment is carried out by exogenously supplementing drugs containing apolipoprotein H or drugs that can promote the expression of apolipoprotein H in the body, thereby reducing alcoholic multi-organ injury.

[0040] Use of a reagent for detecting a biomarker in the preparation of a product for preventing and / or treating alcoholic diseases, wherein the biomarker is apolipoprotein H.

[0041] Although the present invention has been disclosed above by way of examples, it is not intended to limit the present invention. Any person skilled in the art can make various different selections and modifications without departing from the spirit and scope of the present invention. Therefore, the protection scope of the present invention is defined by the claims and their equivalent forms.

Claims

1. Use of apolipoprotein H in the preparation of a medicament or health food for preventing and / or treating alcoholic diseases.

2. The application according to claim 1, wherein The medicament or health food contains apolipoprotein H or the apolipoprotein H gene, and the medicament or health food can maintain the homeostasis of the intestinal microecology and the balance of bile acid metabolism.

3. The application according to claim 1, characterized in that, The medicament or health food is used to alleviate the abnormal bile acid metabolism caused by alcoholic injury, and / or to alleviate the intestinal microecological disorder secondary to alcoholic injury.

4. The application according to claim 1, characterized in that, The alcoholic disease is alcoholic gastrointestinal injury and / or abnormal or imbalanced bile acid metabolism. The alcoholic intestinal injury is intestinal microecological disorder, and the abnormal or imbalanced bile acid metabolism is bile acid metabolism disorder.

5. The application according to claim 1, characterized in that, The medicament or health food further includes a pharmaceutically acceptable excipient. The medicament is an injection, tablet, granule, oral preparation, or genetically engineered drug.

6. Use of apolipoprotein H as a target in the preparation of a medicament for preventing and / or treating alcoholic diseases, characterized in that, The medicament can maintain the homeostasis of the intestinal microecology and the balance of bile acid metabolism.

7. The application according to claim 6, wherein The alcoholic disease is alcoholic gastrointestinal injury and / or abnormal or imbalanced bile acid metabolism. The alcoholic intestinal injury is intestinal microecological disorder, and the abnormal or imbalanced bile acid metabolism is bile acid metabolism disorder.

8. Use of a reagent for detecting a biomarker in the preparation of a product for diagnosing alcoholic diseases, characterized in that, The biomarker is apolipoprotein H.

9. The application according to claim 8, characterized in that, The reagent for detecting the biomarker is a reagent for detecting the expression level of apolipoprotein H.

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