Use of apolipoprotein h in preparation of drug or health-care product for preventing and / or treating alcoholic diseases

Abstract

Provided is the use of apolipoprotein H in the preparation of a drug or a health-care product for preventing and / or treating alcoholic diseases. A 10-week-old ApoH gene-knockout mouse (C57BL / 6ApoH- / -) is used as a background to construct an alcoholic hepatitis model, and an alcoholic hepatitis model in a wild-type C57BL / 6 mouse is used as a control. Transaminase and blood fat levels in peripheral blood and fat content in liver tissue are detected, and immunohistochemical staining of liver tissue is performed. The results show that the TC content in the liver tissue of the ApoH- / -mouse is significantly increased. The composition and content of bile acid in liver tissue and intestinal content, and the composition and abundance of intestinal flora are further detected. The results show that there are abnormal bile acid metabolism and intestinal flora disorders. Further provided is a precise targeting therapeutic drug for treating alcoholic multivisceral injury on the basis of the mechanism that the low expression of the alcoholic apolipoprotein H leads to intestinal flora disorders, affects bile acid metabolism, and simultaneously aggravates hepatocyte steatosis.

Description

Application of apolipoprotein H in the preparation of medicines or health products for preventing and / or treating alcoholic diseases Technical Field

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

[0002] In recent years, the number of patients with alcoholic disorders has increased clinically. From a medical classification perspective, alcoholic disorders can be divided into acute alcoholic disorders and chronic alcoholic disorders. Acute alcoholic disorders are characterized by neurological and intestinal problems that develop after a single heavy drinking session. Chronic alcoholic disorders often occur in patients with chronic alcohol dependence. Due to long-term alcohol dependence, they can experience weakness in the limbs, often accompanied by peripheral nerve problems. They also affect the digestive, circulatory, and endocrine systems, leading to complications such as alcoholic dementia, chronic gastritis, alcoholic fatty liver disease, coronary heart disease, and pancreatitis. Gastrointestinal and liver diseases are particularly common.

[0003] Intestinal dysbiosis is a common gastrointestinal disease. Under certain pathological conditions, the intestinal barrier is disrupted, leading to the translocation of bacteria and their metabolites and abnormal activation of the immune system, causing damage to the intestines and liver.

[0004] Therefore, it is particularly important to develop a drug or health product that can prevent and / or treat alcohol-related diseases.

[0005] Summary of the Invention

[0006] The technical problem to be solved by the present invention is to provide an application of apolipoprotein H in the preparation of medicines or health products for preventing and / or treating alcoholic diseases, which solves the technical problem of alcoholic multi-organ damage.

[0007] The technical problem to be solved by the present invention is achieved through the following technical solutions:

[0008] One object of the present application is to provide a use of apolipoprotein H in the preparation of medicines or health products for preventing and / or treating alcoholic diseases.

[0009] Preferably, the medicine or health product contains apolipoprotein H or an apolipoprotein H gene, and the medicine or health product can maintain intestinal microecological homeostasis and bile acid metabolic balance.

[0010] Preferably, the medicine or health product is used to alleviate bile acid metabolism abnormalities caused by alcoholic injury, and to alleviate intestinal microecological disorders secondary to alcoholic injury.

[0011] Preferably, the alcoholic disease is alcoholic gastrointestinal damage and / or abnormal or imbalanced bile acid metabolism, the alcoholic intestinal damage is intestinal microecological disorder, and the abnormal or imbalanced bile acid metabolism is bile acid metabolism disorder.

[0012] Preferably, the medicine or health product further comprises medically acceptable excipients, and the medicine is an injection, tablet, granule or oral preparation, or a genetically engineered drug.

[0013] The second object of the present application is to provide a use of apolipoprotein H as a target in the preparation of a drug for preventing and / or treating alcoholic diseases, wherein the drug can maintain intestinal microecological homeostasis and bile acid metabolic balance.

[0014] Preferably, the alcoholic disease is alcoholic gastrointestinal damage and / or abnormal or imbalanced bile acid metabolism, the alcoholic intestinal damage is intestinal microecological disorder, and the abnormal or imbalanced bile acid metabolism is bile acid metabolism disorder.

[0015] The third object of the present application is to provide a use of a reagent for detecting a biomarker in the preparation of a product for diagnosing alcoholic diseases, wherein the biomarker is apolipoprotein H.

[0016] Preferably, the reagent for detecting a biomarker is a reagent for detecting the expression level of apolipoprotein H.

[0017] The above technical solution of the present invention has the following beneficial effects:

[0018] This application is based on the mechanism that low expression of alcoholic apolipoprotein H leads to intestinal microecological disorders, affects bile acid metabolism, and aggravates fatty degeneration of hepatocytes, and is a precise targeted therapeutic drug for the treatment of alcoholic multi-organ damage. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 shows the results of liver transcriptome detection and analysis of alcoholic liver disease patients (among which: a is the differential gene analysis of liver transcriptome, b is the change of APOH gene expression in liver, c is the enrichment analysis of alcoholic hepatitis and bile acid metabolic pathway, d is the enrichment analysis of apolipoprotein H expression change and bile acid metabolic pathway).

[0020] Figure 2 shows the results of transcriptome analysis of the liver of a gene-knockout mouse alcoholic fatty liver model (where: a is the expression of the mouse ApoH gene, b is the enrichment analysis of the cholesterol metabolism pathway in the liver of the model mouse, and c is the cholesterol metabolism-related genes).

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

[0022] Figure 4 shows the results of intestinal flora detection and association analysis with bile acid (where: a is the change in intestinal flora abundance, b is the flora related to bile acid metabolism, c is the flora functional 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 DESCRIPTION

[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 arrangement of components and steps, numerical expressions and numerical values ​​set forth in these embodiments do not limit the scope of the present invention.

[0024] The products and technologies used in the following examples of this application are deemed to be conventional operations unless otherwise specified.

[0025] Alcohol intake directly leads to intestinal microecological disorders. It is currently believed that intestinal microecology and bile acid metabolism have a mutual influence. Abnormal bile acid metabolism is one of the main regulatory factors of alcoholic diseases, and the gut-liver axis plays an important regulatory role in the pathogenesis of alcoholic diseases.

[0026] The study showed that 10-week-old ApoH knockout mice (C57BL / 6 ApoH - / - ) was used as the background to construct an alcoholic hepatitis model (chronic-plus-binge model), and the wild-type C57BL / 6 mouse alcoholic hepatitis model was used as the control. The levels of peripheral blood transaminases and blood lipids, the fat content of liver tissue, and the immunohistochemical staining of liver tissue were detected. The results showed that ApoH - / - The TC content in the liver tissue of mice was significantly increased. Further testing of the bile acid composition and content of liver tissue and intestinal contents, as well as the composition and abundance of intestinal flora, showed that there were abnormal bile acid metabolism and intestinal microecological disorders. This application is based on the mechanism that low expression of alcoholic apolipoprotein H leads to intestinal microecological disorders, affects bile acid metabolism, and aggravates hepatocyte fatty degeneration, and is a precise targeted therapeutic drug for the treatment of alcoholic multi-organ damage.

[0027] Example 1 Analysis of Apolipoprotein H and Alcoholic Diseases

[0028] Figure 1 shows the results of liver transcriptome analysis in patients with alcoholic liver disease. As shown in Figure 1, we found that apolipoprotein H gene expression was significantly reduced in patients with alcoholic hepatitis, with significant differences compared to healthy controls (Figure 1, Panels a and b). Patients with alcoholic hepatitis exhibit abnormal bile acid metabolism (Figure 1, Panel c). Further analysis revealed that changes in apolipoprotein H expression are closely correlated with bile acid metabolic activity: those with relatively high apolipoprotein H expression have enhanced bile acid synthesis capacity, while those with relatively low apolipoprotein H expression have significantly reduced bile acid transport and secretion capacity (Figure 1, Panel d). The regulatory mechanisms of apolipoprotein H and alcoholic diseases along the gut-liver axis remain unclear, so a mouse model was constructed for further investigation.

[0029] Knockout mice were constructed. This method can be referenced to existing technologies and will not be described in detail here. A chronic-plus-binge ethanol-fed mouse model of alcoholic hepatitis was established in the APOH knockout mouse background. Non-alcohol-fed knockout mice served as controls. Serum transaminases, blood lipids, liver tissue fat content, and liver tissue pathological staining were used to characterize the model's phenotype and changes.

[0030] The results showed that alcohol-fed knockout mice exhibited increased liver cholesterol and aggravated steatosis. Further liver transcriptome analysis analyzed differential gene expression and functional enrichment. The results suggest that differentially expressed genes in the alcohol-fed knockout mice are enriched in cholesterol metabolism pathways, including bile acid synthesis and secretion (Figure 2). Therefore, we hypothesize that the alcohol-fed knockout mice are involved in regulating bile acid metabolism.

[0031] Furthermore, the bile acid content in the liver tissue and intestinal contents of the mice was further tested, and analysis revealed significant differences in the composition and content of bile acids in the liver tissue and intestinal contents (as shown in Figure 3). In the liver tissue of the alcohol-treated knockout mice, the composition ratio of taurocholic acid (TCA) and hyocholic acid (HCA) was significantly increased, and the content of taurine-conjugated secondary bile acids was significantly increased, as was the content of cholic acid (CA) and hyocholic acid (HCA). Furthermore, in the intestinal contents, the composition ratio and content 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 also that the enterohepatic circulation of bile acids is affected.

[0032] Furthermore, changes in the intestinal flora of the model mice were examined. Through diversity and abundance analysis, as well as differential bacterial flora functional prediction, it was found that the abundance and composition of the intestinal flora in the alcohol-induced knockout mice were significantly different (Figure 4a, b). Functional prediction analysis suggested that the differential bacterial flora were associated with metabolic function (Figure 4c). Furthermore, correlation analysis with bile acid content (Figure 4d) revealed bacterial flora associated with changes in bile acid content.

[0033] In summary, alcoholic low APOH expression can directly lead to intestinal microecological disorders, affect bile acid metabolism, and then aggravate liver steatosis. Abnormal bile acid metabolism is one of the main regulatory factors of alcoholic multi-organ damage.

[0034] Example 2 Alcoholic APOH High Expression

[0035] 1. In 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 the control. Peripheral blood, intestinal tissue, and intestinal contents were collected from mice at different time points after injection. The composition and abundance changes of the intestinal flora of the model mice were examined, as well as the enrichment of differential bacterial functions, changes in bile acid metabolism, and the correlation analysis between bile acid changes and intestinal flora. Pathological staining of liver and intestinal tissues was used to identify the degree of organ damage.

[0036] 2. Targeting the mouse liver to overexpress the ApoH gene, further constructing an alcohol-feeding model, collecting peripheral blood, liver and intestinal tissues, and intestinal contents from mice at different times, examining the intestinal flora composition, abundance changes, differential flora functional enrichment, bile acid metabolism changes, and the correlation between bile acid changes and intestinal flora. Pathological staining of liver and intestinal tissues was performed to identify the degree of organ damage.

[0037] 3. In the context of the above-mentioned alcohol-fed mouse model, oral antibiotics were given to construct intestinal germ-free mice. The antibiotic group was used as a control. Peripheral blood and liver tissues of mice were collected at different time points to detect changes in the expression of APOH genes and proteins to further clarify the direct correlation between APOH expression and changes in intestinal flora.

[0038] The results showed that high APOH expression can restore homeostasis to alcohol-induced intestinal microecological disturbances and alleviate abnormalities in bile acid metabolism. This further suggests that detecting APOH levels and increasing APOH levels to maintain normal levels are important for the prevention and treatment of alcohol-induced intestinal microecological disturbances and multi-organ damage.

[0039] Research on apolipoprotein H has revealed that it can serve as a detection target for alcoholic disorders, making their diagnosis and treatment easier. Treatment can be achieved by exogenously supplementing with drugs containing apolipoprotein H or by supplementing with drugs that promote apolipoprotein H expression in the body, thereby alleviating alcohol-induced multi-organ damage.

[0040] Use of a reagent for detecting a biomarker in preparing 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 embodiments, they are not intended to limit the present invention. Any person skilled in the art may make various choices and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention is defined by the claims and their equivalents.

Claims

1. Use of apolipoprotein H in the preparation of medicines or health products for preventing and / or treating alcoholic diseases.

2. The use according to claim 1, characterized in that The medicine or health product contains apolipoprotein H or an apolipoprotein H gene, and the medicine or health product can maintain intestinal microecological homeostasis and bile acid metabolic balance.

3. The use according to claim 1, characterized in that The medicine or health product is used to alleviate bile acid metabolism abnormalities caused by alcoholic injury, and / or to alleviate intestinal microecological disorders secondary to alcoholic injury.

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

5. The use according to claim 1, characterized in that The medicine or health product also includes medically acceptable excipients, and the medicine is an injection, tablet, granule or oral preparation, or a genetically engineered medicine.

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

7. The use according to claim 6, characterized in that The alcoholic disease is alcoholic gastrointestinal damage and / or abnormal or imbalanced bile acid metabolism, the alcoholic intestinal damage 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 use according to claim 8, characterized in that The reagent for detecting biomarkers is a reagent for detecting the expression level of apolipoprotein H.

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