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Medicine and treatment method for treating hepatic fibrosis

A liver fibrosis and drug technology, applied in the field of drugs targeting NS5ATP9 gene screening for treatment or prevention of tissue fibrosis, can solve the problem of no effective treatment methods

Inactive Publication Date: 2018-10-23
北京泛亚同泽生物医学研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no effective treatment for liver fibrosis or cirrhosis caused by other factors such as alcohol, metabolism, drugs, etc. Traditional Chinese medicine or Chinese patent medicine is mainly used as adjuvant treatment

Method used

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  • Medicine and treatment method for treating hepatic fibrosis
  • Medicine and treatment method for treating hepatic fibrosis
  • Medicine and treatment method for treating hepatic fibrosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1: Differential expression of NS5ATP9 in tissues with different degrees of liver fibrosis

[0052] Liver biopsies from 24 patients with HBV-related liver fibrosis were obtained from the Affiliated Hospital of Putian University, Fujian Province. After routine pathological H&E staining and Masson staining (eg figure 1 Shown in A-C), according to the Knodell HAI scoring system, the degree of liver fibrosis was staged and divided into three groups: S0~S1, S2~S3, and S4. Immunohistochemical staining method was used to detect the expression level of NS5ATP9 protein in liver biopsy tissues of the above three groups. According to the semi-quantitative analysis method reported by Chung et al., the results showed that in the three groups of patients with different degrees of fibrosis, there were significant differences in the expression level of NS5ATP9 and the number of positive staining cases. , its expression decreased significantly, and the difference was statistica...

Embodiment 2

[0053] Example 2: Regulatory relationship between NS5ATP9 gene expression level and liver fibrosis level

[0054] 1. Carbon tetrachloride (CCl 4 ) to induce the establishment of mouse liver fibrosis model:

[0055] 1) A total of 24 C57BL / 6 mice were randomly divided into experimental group and control group, with 12 mice in each group, and were treated as follows:

[0056] Experimental group mice (n=12): intraperitoneal injection of CCl 4 -Olive oil mixed solution (volume ratio 1:4), 2.5 μl / g, 2 times / week; control group mice (n=12): intraperitoneal injection of olive oil solution, 2.5 μl / g, 2 times / week.

[0057] 2) On the second day after the end of modeling 2 weeks and 4 weeks respectively, 1% pentobarbital solution was injected intraperitoneally, the mice were anesthetized and sacrificed, fresh liver tissues were taken, and part of them were fixed in 4% paraformaldehyde solution. The other part was stored in liquid nitrogen for future use.

[0058] 3) Carry out paraffi...

Embodiment 3

[0075] Example 3: Expression of NS5ATP9 gene induces apoptosis of hepatic stellate cells and inhibits proliferation of hepatic stellate cells

[0076] 1. The effect of NS5ATP9 on the apoptosis of hepatic stellate cells (HSC) was analyzed by flow cytometry Annexin V-FITC / 7-AAD cell apoptosis detection technology:

[0077] 1) Experimental method:

[0078] Apoptosis of activated HSC is considered to be an important formation mechanism in the process of liver fibrosis reversal and ECM degradation. Therefore, we used flow cytometry Annexin V-FITC / 7-AAD cell apoptosis detection technology to analyze the effect of NS5ATP9 on HSC apoptosis. role. pcDNA3.1 / myc-His9(-)-NS5ATP9 overexpression plasmid (pcDNA3.1 / myc-His(-)-NC as negative control) / siRNA-NS5ATP9 (siRNA-NC as negative control) was transiently transfected into untreated In the stimulated LX-2 cells, the changes of apoptosis were detected 48 hours after transfection.

[0079] 2) Results:

[0080] After overexpression of NS5...

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Abstract

The invention relates to a screening method for a medicine for targeting NS5ATP9 (HCV NS5A-transactivated protein 9) prevention or treatment of tissue fibrosis and tissue sclerosis, and the medicine obtained by screening using the method. NS5ATP9 significantly regulates the expression of extracellular matrix collagen and non-collagen glycoprotein and affects the occurrence and development of tissue fibrosis by regulating a TGFbeta1 / Smad3 signaling pathway; and the NS5ATP9 promotes the apoptosis of hepatic stellate cells, inhibits the proliferation of the hepatic stellate cells and regulates extracellular matrix deposition by regulating the phosphorylation level and the intracellular translocation condition of a Smad3 protein. TDF / TAF can effectively regulate the expression of NS5ATP9 gene,and is verified in CCl4-induced mouse liver fibrosis model, and the TDF / TAF achieves significant effects in the prevention or treatment of CCl4-induced liver fibrosis in mice.

Description

technical field [0001] The invention belongs to the field of pharmacy, and in particular, the invention relates to a method for targeting NS5ATP9 gene to screen a drug for treating or preventing tissue fibrosis, and the drug obtained by screening through the method. Background technique [0002] Fibrosis can occur in a variety of tissues and organs. The main pathological changes are the increase of fibrous connective tissue and the reduction of parenchymal cells in organ tissues. Continuous progress can lead to organ structural damage, functional decline, and even failure, which seriously threaten human health and life. Worldwide, tissue fibrosis is the main cause of disability and death in many diseases. According to relevant statistics in the United States, nearly 45% of the patients who died of various diseases in the United States can be attributed to tissue fibroproliferative diseases. [0003] Liver fibrosis is especially common in patients with tissue fibrosis. It is ...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K31/675A61K49/00A61P1/16A01K67/02C12N15/90
CPCA01K67/02A61K31/675A61K45/00A61K49/0008C07K14/005C12N9/22C12N15/907C12N2760/16322A61K31/18A61K31/20A61K31/7084A61K49/00A61P1/16C12N15/85C12N15/90
Inventor 成军
Owner 北京泛亚同泽生物医学研究院有限公司
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