Picric acid-sirius red staining method and application thereof in screening anti-hepatic fibrosis compound

Abstract

The invention belongs to biotechnology field and particularly relates to high throughput screening of an effective anti-hepatic fibrosis compound by means of picric acid-sirius red staining method on a collagenous fiber. The cell is hepatic stellate cell line CFSC-8B, transforming growth factor TGF-beta1 with the dose of 10ng / mL can obviously induce the CFSC-8B cell to secrete collagen when acting for 48 hours, and the secretion of the CFSC-8B cell collagen induced by the TGF-beta 1 can be effectively detected by means of picric acid-sirius red staining method. The picric acid-sirius red staining method on the collagenous fiber is applied to screening the compounds with anti-hepatic fibrosis activities in different polarity extracts of antrodia camphorata, ganoderma lucidum, cephalosporium sinensis, cordyceps mortierella, armillaria and hericium erinaceus powder. Tests prove that n-hexane and chloroform extract of the antrodia camphorata, ethyl acetate extract from the cordyceps mortierella, and n-hexane, chloroform extract and ethyl acetate extract from the armillaria can effectively inhibit the secretion of the CFSC-8B cell collagen induced by the TGF-beta 1, the above extracts have dose dependent relationship, are significant in difference, and have an anti-hepatic fibrosis function, and other extracts do not have significant inhibition effect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to high-throughput screening of effective compounds for anti-hepatic fibrosis in vitro by using the picric acid sirius red staining method of collagen fibers. Background technique [0002] Liver fibrosis (Liver fibrosis) refers to the pathological process of excessive precipitation of diffuse extracellular matrix (ECM) in the liver caused by various pathogenic factors during the pathogenesis of many chronic liver diseases. ECM includes collagen, fiber Connexin, elastin, laminin, hyaluronic acid and proteoglycan, etc. Hepatic stellate cells (HSCs) are the main source of collagen and other extracellular matrix in the liver. Activated stellate cells express α-smooth muscle actin (alpha-smooth muscle actin, α-SMA), secrete collagen types I, III, and IV, matrix metalloproteinases (matrix metalloproteinases, MMPs) and metalloproteinase inhibitors (tissue inhibitor of metalloprot...

AI Technical Summary

Problems solved by technology

According to relevant literature reports, the picric acid Sirius red staining method is applied to the quantitative analysis of collagen fibers in the heart and kidney, the study of alcoholic liver fibrosis, and the evaluation of pancreatic fibrosis in rats, but the picric acid Sirius red staining method is mostly applied For the staining of animal tissue sections, due to the long cycle of animal experiments and the instability of experimental results due to individual differences, it is difficult to achieve the purpose of high-throughput screening of anti-hepatic fibrosis compounds in vivo experiments

Picric acid Sirius scarlet staining has not been reported yet on the use of multi-well plates for high-throughput screening of anti-hepatic fibrosis compounds at the animal cell level

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