CRISPR/Cas9 silencing Fas inhibits autophagy formation and mitochondrial damage in acute liver failure cells

doi:10.19871/j.cnki.xfcrbzz.2023.03.002

Abstract

Abstract: Objective In order to explore the role and mechanism of the death receptor Fas/CD95 in the progression of acute liver failure. Method We first established, a mouse animal model of ALF induced by concanavalin A (Con A). The formation of mitophagy in hepatocyte with ALF was detected by Western blot and transmission electron microscopy. Then, Knocking out Fas by administering CRISPR/Cas9 plasmid through the tail vein of mice,and then acute liver failure of mouse was induced by Con A, liver tissue injury, the formation of mitophagy and mitochondrial damage were detected. We treated HepG2 that knocked out Fas with Con A in vitro to detect the expression of mitochondrial autophagy related proteins and mitochondrial damage. Result The results show that after acute liver failure induced by concanavalin A in mice, transmission electron microscopy observation revealed mitochondrial autophagy in liver cells, a decrease in p62 protein levels, upregulation of PINK1/Parkin protein, and upregulation of Fas expression. After transfection with PX330Fas plasmid, the expression ratio of LC3II/LC3I in mouse liver tissue and in vitro liver cells decreased, the expression of p62 protein was restored, the levels of mitochondrial protein COX4I1 and mitochondrial DNA content decreased, and Fas expression was downregulated, with significant statistical significance. Conclusion In this study, we found that CRISPR/Cas9 silencing Fas inhibits the progression of ALF, the formation of mitochondrial autophagy and mitochondrial damage in hepatocytes.

Key words: Acute liver failure, Mitochondria, Autophagy, Death receptor Fas/CD95, Mitochondrial damage

CLC Number:

R575.3

Wu Zhijing, Zhang Wenyuan, Xu Zhiqi, Zhuang Peng. CRISPR/Cas9 silencing Fas inhibits autophagy formation and mitochondrial damage in acute liver failure cells[J]. Electronic Journal of Emerging Infectious Diseases, 2023, 8(3): 7-11.

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