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Application of osteoprotegerin only or combination of osteoprotegerin with other cytokines in treatment of hepatic fibrosis

A technology of osteoprotegerin and liver fibrosis, which is applied in medical preparations containing active ingredients, digestive system, drug combination, etc., and can solve the problems of short and long tracking time.

Active Publication Date: 2016-09-21
ZHEJIANG SHENGCHUANG PRECISION MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although malignant transformation of human MSCs has not been reported in clinical trials, it is also possible that follow-up time is relatively short, whereas tumorigenesis may take longer to emerge

Method used

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  • Application of osteoprotegerin only or combination of osteoprotegerin with other cytokines in treatment of hepatic fibrosis
  • Application of osteoprotegerin only or combination of osteoprotegerin with other cytokines in treatment of hepatic fibrosis
  • Application of osteoprotegerin only or combination of osteoprotegerin with other cytokines in treatment of hepatic fibrosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Establishment and Identification of Liver Fibrosis Mouse Model

[0031] The present invention uses commonly used in this field through CCl 4 A mouse model of liver fibrosis established by injuring the mouse liver.

[0032] 6-8 week ICR mice (purchased from Shanghai SLAC Experimental Animal Co., Ltd.) were intraperitoneally injected with CCl 4 , according to 1mL CCl 4 / kg mouse body weight injection. CCl 4 Dissolved in olive oil (olive oil) at a ratio of 10%, injected twice a week with a 1 mL micro-syringe (BD Biosciences) for 4 weeks, thus establishing an early liver fibrosis model in mice. By removing the mouse liver and staining it with sirius red, the histopathological conditions of the liver were examined; at the same time, the serum liver function indicators of the mice were detected, including ALT, AST, ALB, ALP and TBIL. Through the above indicators, it was verified whether the mouse fibrosis model was successfully constructed.

Embodiment 2

[0033] Example 2: Isolation and in vitro culture of MenSCs

[0034] During the menstrual cycle of menstrual healthy female volunteers, female menstrual blood samples were collected using Divacup (Kitchener, ON). Samples were carefully stored at 4°C within 2 days of harvesting. After these samples are taken out, the subsequent operating procedures are processed in the cell room. The collected menstrual blood was centrifuged (1000rpm, 10min, 4°C), separated by Ficoll-Paque to obtain mononuclear cells (mixed type) in the liquid, and then the cell samples were tested to detect the number of bacteria in the supernatant. After reaching the standard, the cells were cultured in vitro. After cell separation, wash 3-5 times with PBS (discard impurities as much as possible), then transfer the isolated and purified cells to DMEM / F12 medium (added with 1% penicillin / streptomycin, 1% amphotericin B and 15% FBS ) in cell culture flasks (Corning) for continued adherent culture. The separa...

Embodiment 3

[0037] Example 3: Construction and screening of MenSCs (GFP+MenSCs) expressing green fluorescent protein (GFP)

[0038] P2-P3 MenSCs were cultured in T75 culture flasks (Corning). When the cells adhered to the wall and grew to about 50%, the medium was discarded, washed 3 times with PBS, and polybrene containing 6 μg / mL (Hanbio Technology Co., Ltd.) was added. ) and GFP-PURO (Hanheng Biological Technology Co., Ltd.) virus liquid (MOI=50; MOI, multiplicity of infection) 10mL Chang's complete medium (Hangzhou Yiwensai Biotechnology Co., Ltd.). After 1 day, the fresh medium was replaced, and after the cells were cultured for 2-3 days, the fresh medium was replaced again, and 2 μg / mL puromycin (Sigma) was used for cell selection. The screened cells (GFP+MenSCs) were tested positive for GFP and expressed and identified cell surface marker molecules (including CD29, CD34, CD45, CD73, CD90, CD105, CD117 and HLA-DR).

[0039] Some of the cells after two weeks of screening were taken ...

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Abstract

The invention relates to application of osteoprotegerin (OPG) only or combination of osteoprotegerin with other cytokines in treatment of hepatic fibrosis. Osteoprotegerin only or combination of osteoprotegerin with other cytokines can improve the condition of the hepatic fibrosis. Therefore, the hepatic fibrosis is better cured, or another feasible selection is provided for treating the hepatic fibrosis, and the prognosis or life quality of patients is improved.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals, and specifically relates to the use of osteoprotegerin (OPG) alone or in combination with other cytokines in the treatment of liver fibrosis. Background technique [0002] Liver disease seriously affects human health and quality of life, and is one of the major infectious diseases facing human beings. Tens of thousands of patients in China die of various liver diseases every year. Because of the high number of complications and high mortality, the treatment of liver diseases has attracted widespread attention and research all over the world. Liver diseases mainly include liver injury, fulminant liver failure, liver fibrosis, cirrhosis, hepatocellular carcinoma, cholestatic liver, autoimmune liver disease, alcoholic fatty liver and non-alcoholic fatty liver, etc. Liver fibrosis is a wound-injury response to various acute / chronic stimuli (including alcohol, viral infections, drugs, toxins, bile, ...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61K38/19A61K38/20A61K38/18A61P1/16
CPCA61K38/1793A61K38/1833A61K38/195A61K38/204A61K38/2053A61K2300/00
Inventor 余艳春
Owner ZHEJIANG SHENGCHUANG PRECISION MEDICAL TECH CO LTD
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