CN116603053A - Use of polypeptide in preparing medicine for preventing and treating non-alcoholic fatty liver disease - Google Patents

Abstract

The invention belongs to the field of medicines, and in particular relates to application of a polypeptide in preparing a medicine for preventing and treating non-alcoholic fatty liver disease. The invention provides application of polypeptide Ac2-26 in preparing a medicine for preventing and treating non-alcoholic fatty liver disease, and particularly has good treatment effect on non-alcoholic steatohepatitis (NASH). The invention discovers that the polypeptide Ac2-26 can obviously reduce lipid deposition, inflammation and fibrosis of liver, has definite therapeutic effect on nonalcoholic steatohepatitis, provides a new choice for clinical medication, and has great clinical and social significance.

Description

Use of polypeptide in preparing medicine for preventing and treating non-alcoholic fatty liver disease

technical field

The invention belongs to the field of medicine, and in particular relates to the use of polypeptides in the preparation of medicines for preventing and treating nonalcoholic fatty liver disease.

Background technique

Nonalcoholic fatty liver disease (NAFLD) refers to a clinicopathological syndrome characterized by diffuse hepatic bullous steatosis caused by causes other than alcohol and other definite liver injury factors. With the significant increase in the incidence of obesity and type II diabetes, NAFLD has become the main cause of chronic liver disease, and its global incidence is about 25.24%. According to clinical and pathological features, NAFLD is usually divided into simple nonalcoholic fatty liver (NAFL) and nonalcoholic steatohepatitis (NASH). NASH is a chronic liver disease characterized histologically by hepatic steatosis, inflammatory cell infiltration or interstitial fibrous proliferation, and is an important pathological stage of NAFLD. Compared with NAFL, NASH patients can further develop end-stage liver diseases such as liver fibrosis, liver cirrhosis, and hepatocellular carcinoma.

The pathogenesis of NASH is complex, involving complex interactions among lipid deposition, inflammation, oxidative stress, fibrosis, abnormal bile acid metabolism, gut microbiota dysbiosis, and susceptibility gene variation. Therefore, the pathogenesis of NASH has not been fully elucidated, and there is no effective drug approved for the treatment of NASH.

Contents of the invention

Aiming at the lack of effective therapeutic drugs for non-alcoholic fatty liver disease (NAFLD) in the prior art, the present invention provides the use of polypeptide Ac2-26 in the preparation of drugs for preventing and treating non-alcoholic fatty liver disease.

Further, the nonalcoholic fatty liver disease is simple nonalcoholic fatty liver (NAFL) or nonalcoholic steatohepatitis (NASH).

Preferably, the nonalcoholic fatty liver disease is nonalcoholic steatohepatitis (NASH).

The invention provides the use of the polypeptide Ac2-26 in the preparation of a medicine for reducing lipid deposition, inflammation or fibrosis in the liver.

The invention also provides the use of the polypeptide Ac2-26 in the preparation of medicines for preventing and treating liver cirrhosis or hepatocellular carcinoma.

Further, the drug reduces liver lipid deposition.

Further, the drug reduces liver inflammation.

Further, the drug reduces the expression of liver fibrosis genes and proteins.

Further, the drug reduces the deposition of collagen fibers in the liver.

Further, the amino acid sequence of the polypeptide is SEQ ID NO.1.

SEQ ID NO. 1: AMVSEFLKQAWFIENEEQEYVQTVK.

Further, the medicine is a preparation prepared by taking the polypeptide Ac2-26 as the active ingredient and adding pharmaceutically acceptable excipients or auxiliary ingredients.

Further, the preparation is an oral preparation, a nasal administration preparation or an injection preparation.

Further, the unit preparation contains 0.0.1-0.5 mg of the polypeptide Ac2-26. Preferably, the unit preparation contains 0.0813 mg, 0.1626 mg, and 0.3252 mg of the polypeptide Ac2-26. The unit preparation of the present invention corresponds to the dosage per kg body weight of the human body per day.

In the examples, the doses of Ac2-26 polypeptide given to mice are 1, 2, 4 mg/kg/monkey/day, and the converted adult doses are 0.0813, 0.1626, 0.3252 mg/kg/person/day.

Beneficial effects: the invention provides the use of the polypeptide Ac2-26 in the preparation of medicines for preventing and treating non-alcoholic fatty liver disease, especially having a good therapeutic effect on non-alcoholic steatohepatitis (NASH). The present invention finds that the polypeptide Ac2-26 can significantly reduce lipid deposition, inflammation and fibrosis in the liver, has a definite therapeutic effect on non-alcoholic steatohepatitis, provides a new option for clinical medication, and has a large clinical and social significance.

Description of drawings

Fig. 1 is the liver lipid quantitative determination and Oil Red O staining result figure of embodiment 1 of the present invention;

Fig. 2 is the results of detection of liver inflammation-related genes and H&E staining in Example 2 of the present invention;

Figure 3 is a diagram showing the results of liver fibrosis-related gene detection and protein detection in Example 3 of the present invention;

Fig. 4 is a graph showing the results of liver Sirius red staining and Masson staining in Example 4 of the present invention.

Detailed ways

The solutions of the present invention will be explained below in conjunction with examples. Those skilled in the art will understand that the following examples are only for illustrating the present invention and should not be considered as limiting the scope of the present invention. If no specific technique or condition is indicated in the examples, it shall be carried out according to the technique or condition described in the literature in this field or according to the product specification. The reagents or instruments used were not indicated by the manufacturer, and they were all commercially available conventional products.

Ac2-26 polypeptide (commercially available product), the sequence of which is SEQ ID NO.1: AMVSEFLKQAWFIENEEQEYVQTVK, was prepared into Ac2-26 polypeptide solution with physiological saline.

Experimental method: C57BL/6 mice were divided into 5 groups on average, namely: 1) normal control group; 2) model group; 3) Ac2-26 low-dose group; 4) Ac2-26 medium-dose group; 5) Ac2- 26 high dose group. The mice in the normal control group were fed with MCS feed, and the model group and Ac2-26 group were fed with methionine choline deficiency (MCD) feed. Inject 1, 2, 4 mg/kg of Ac2-26 polypeptide solution once a day. After 4 weeks, the mice were sacrificed, and the liver tissues were taken for embedding and sectioning, and then stained with H&E, Oil red O, Sirius red and Masson. Lipid was extracted from liver tissue homogenate, and the lipid content in the liver was quantitatively detected. At the same time, RNA and protein were extracted from the liver tissue, and the expressions of inflammation and fibrosis-related genes and proteins were detected by RT-PCR and Western blot.

Embodiment 1Ac2-26 treatment reduces hepatic lipid deposition

Liver lipids were extracted, and the contents of triglyceride (TG), total cholesterol (TC) and free fatty acid (NEFA) in the liver were detected. The results are shown in Figure 1. It can be seen from Figure 1A that the contents of TG, TC, and NEFA were significantly increased after MCD modeling, and liver lipids were reduced to varying degrees after Ac2-26 treatment, which was obviously dose-dependent. Oil red O staining of frozen liver sections showed that liver lipid droplets increased significantly after MCD modeling, and liver lipid droplets decreased in a dose-dependent manner after Ac2-26 treatment (Fig. 1B).

Embodiment 2Ac2-26 treatment reduces liver inflammation

Liver mRNA was extracted, and the expression of liver inflammatory factors was detected by RT-PCR. The results showed that MCD modeling significantly increased the expression of mRNA levels of liver inflammation-related genes Mcp-1, Tnf-α, Cd68 and TGF-β, and the administration of Ac2-26 could dose-dependently inhibit the increase of liver inflammation caused by MCD (FIG. 2A). H&E staining results showed that MCD modeling significantly increased the infiltration of inflammatory cells in the liver, and Ac2-26 treatment could improve liver inflammation in a dose-dependent manner (Fig. 2B).

Example 3 Ac2-26 treatment reduces the expression of liver fibrosis genes and proteins

Liver mRNA was extracted, and RT-PCR was used to detect the expression of liver fibrosis-related genes. The results showed that MCD modeling significantly increased the expression of mRNA levels of liver fibrosis-related genes, and the administration of Ac2-26 could dose-dependently inhibit the increase of liver fibrosis-related genes caused by MCD (Fig. 3A). Western blot results showed that MCD modeling significantly increased the expression of liver fibrosis-related proteins alpha-SMA and Col1a1, and Ac2-26 treatment could reduce the expression of fibrosis proteins in a dose-dependent manner (Fig. 3B).

Embodiment 4Ac2-26 treatment reduces hepatic collagen fiber deposition

Sirius red staining and Masson staining were performed on the liver sections, and the results showed that MCD modeling significantly increased the deposition of collagen fibers in the liver, and Ac2-26 treatment could reduce the deposition of collagen fibers in a dose-dependent manner (Figure 4).

It should be noted that the specific features, structures, materials or characteristics described in this specification may be combined in any one or more embodiments in a suitable manner. In addition, those skilled in the art can combine and combine different embodiments and features of different embodiments described in this specification without conflicting with each other.

Claims (10)

1. Use of polypeptide Ac2-26 in the preparation of medicines for preventing and treating non-alcoholic fatty liver disease. 2. The use according to claim 1, characterized in that: the nonalcoholic fatty liver disease is simple nonalcoholic fatty liver or nonalcoholic steatohepatitis. 3. The use according to claim 2, characterized in that: the nonalcoholic fatty liver disease is nonalcoholic steatohepatitis. 4. Use of the polypeptide Ac2-26 in the preparation of a medicament for reducing lipid deposition, inflammation or fibrosis in the liver. 5. Use of the polypeptide Ac2-26 in the preparation of medicines for preventing and treating liver cirrhosis or hepatocellular carcinoma. 6. The use according to any one of claims 1-5, characterized in that: the drug satisfies at least one of the following: The drug reduces hepatic lipid deposition; The drug reduces liver inflammation; The drug reduces the expression of liver fibrosis genes and proteins; The drug reduces collagen fiber deposition in the liver. 7. The use according to any one of claims 1-5, characterized in that: the amino acid sequence of the polypeptide is shown in SEQ ID NO.1. 8. The use according to any one of claims 1-5, characterized in that: the drug is a preparation prepared by taking the polypeptide Ac2-26 as the active ingredient and adding pharmaceutically acceptable excipients or auxiliary ingredients. 9. The use according to claim 8, characterized in that: the preparation is an oral preparation, a nasal cavity administration preparation or an injection preparation. 10. The use according to claim 9, characterized in that: the unit preparation contains 0.01-0.5 mg of the polypeptide Ac2-26.