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Method for screening protein biological markers related to hepatic stellate cell activation and hepatic fibrosis

A hepatic stellate cell and biomarker technology, applied in chemical instruments and methods, peptide preparation methods, organic chemistry, etc., can solve the problems of difficult separation and purification of primary hepatic stellate cells, large sample protein requirements, etc. To achieve the effect of easy operation and simple method

Inactive Publication Date: 2012-08-01
NANTONG UNIVERSITY
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Problems solved by technology

[0002] Screening of hepatic stellate cell activation-related proteins helps to understand the molecular mechanism of liver fibrosis, but because the traditional two-dimensional gel electrophoresis (2-DE)-based protein separation and identification technology requires a large amount of sample proteins, and primary liver It is difficult to separate and purify stellate cells. At present, only Kristensen's research group has used the method of two-dimensional gel electrophoresis and silver staining to identify and compare the protein expression changes of primary rat hepatic stellate cells before and after activation.

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  • Method for screening protein biological markers related to hepatic stellate cell activation and hepatic fibrosis
  • Method for screening protein biological markers related to hepatic stellate cell activation and hepatic fibrosis
  • Method for screening protein biological markers related to hepatic stellate cell activation and hepatic fibrosis

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Embodiment Construction

[0041] A method for screening hepatic stellate cell activation and liver fibrosis-related protein biomarkers, comprising the following steps:

[0042] (1) Auto-activation model of primary hepatic stellate cells cultured in vitro: primary hepatic stellate cells were stained with 0.04% trypan blue for cell count and viability detection, and then treated with high-glucose DMEM containing 10% fetal bovine serum Dilute the complete medium and adjust the cell density to 10 6Cells / ml, inoculated in cell culture flasks or culture plates, cultured in a 37°C incubator containing 5% CO2 and 95% air, changed the medium the next day, and changed the medium every 48hr thereafter, and passaged in 3-4 days; The autofluorescence of vitamin A lipid droplets in the cytoplasm of hepatic stellate cells was observed by UV excitation wavelength, Sudan III fat staining and immunofluorescence were used to detect desmin, a specific marker of rat hepatic stellate cells, and the obtained primary hepatic ...

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Abstract

The invention discloses a method for screening protein biological markers related to hepatic stellate cell activation and hepatic fibrosis. The method comprises the preparation of the automatic aviation models of hepatic stellate cells subjected to isolation culture and in vitro culture and the preparation of static and activated hepatic stellate cell protein samples. The invention has the advantages that the method is simple, and the operation is easy.

Description

technical field [0001] The invention relates to a method for screening protein biomarkers related to activation of hepatic stellate cells and liver fibrosis. Background technique [0002] Screening of hepatic stellate cell activation-related proteins helps to understand the molecular mechanism of liver fibrosis, but because the traditional two-dimensional gel electrophoresis (2-DE)-based protein separation and identification technology requires a large amount of sample proteins, and primary liver It is difficult to separate and purify stellate cells. At present, only Kristensen's research group has used the method of two-dimensional gel electrophoresis and silver staining to identify and compare the protein expression changes of rat primary hepatic stellate cells before and after activation. Due to the limitations of the method, Kristensen et al. identified about 150 proteins in total, 43 of which had significant changes in the expression of activated hepatic stellate cells....

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/18C07K1/14G01N27/62
Inventor 季菊玲季煜华季秋虹鞠剑峰陆锦标刘海鸥陆鹏姚婵
Owner NANTONG UNIVERSITY
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