Effect of inhibition of heat shock protein Gp96 expression on alcoholic liver fibrosis in mice by CRISPR/Cas-9 technique

ZHU Wen-Feng; LI San-Qiang; SONG Xiao-Gai; GUO Wei; YANG Huan; ZHANG Bing-Bing

doi:10.16098/j.issn.0529-1356.2021.05.016

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Acta Anatomica Sinica ›› 2021, Vol. 52 ›› Issue (5) : 777-783. DOI: 10.16098/j.issn.0529-1356.2021.05.016

Histology,Embryology and Developmental Biology

Effect of inhibition of heat shock protein Gp96 expression on alcoholic liver fibrosis in mice by CRISPR/Cas-9 technique

ZHU Wen-feng^1,2LI San-qiang^1,2* SONG Xiao-gai^1,2 GUO Wei^1,2 YANG Huan^1,2 ZHANG Bing-bing^1,2#br#

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Abstract

Objective To investigate the effects of heat shock protein Gp96 on alcoholic liver fibrosis in mice. Methods A total of 220 male healthy C57BL/6 J mice were randomly divided into four groups: normal control group (n=10), saline+alcohol induced liver fibrosis group (n=70), the injection of CRISPR expression Gp96-sgRNA3 by tail vein+alcohol induced liver fibrosis group (n=70), the intraperitoneal injection of nuclear factor kappa B(NF-κB) inhibitors PDTC+alcohol induced liver fibrosis group (n=70). The blood was got from eyeballs and the mice were killed after 8 weeks of ethanol induction. We detected the activity of serum aspartate aminotransferase (AST) in mice of different groups. The pathological changes were detected by HE staining, sirius red staining and periodic acid-Schiff (PAS) staining in the liver of mice. The expression of Gp96 and transforming growth factor β1(TGF-β1)were detected by Western blotting. Results Compared with the normal control group, the AST enzyme activity and liver fibrosis increased significantly, glycogen decreased significantly in other three groups (P<0.01). Compared with the saline+alcohol group, the AST enzyme activity and liver fibrosis increased more significantly, glycogen decreased more significantly, Gp96 expression decreased significantly and TGF-β1 expression increased significantly in Gp96-sgRNA3+ alcohol group and NF-κB inhibitors PDTC+alcohol group (P<0.01 or P<0.05). Conclusion The injection of CRISPR expression plasmid Gp96-sgRNA3 by tail vein significantly inhibited the Gp96 expression, promoted the degree of alcoholic liver fibrosis in mice, and NF-κB signaling pathway played a certain role in regulating the expression of Gp96.

Key words

Heat shock protein Gp96 / Alcohol induced liver fibrosis / Nuclear factor κB signal pathway / Clustered regularly interspaced short palindromic repeats/associated protein 9 / Western blotting / Mouse

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ZHU Wen-feng LI San-qiang SONG Xiao-gai GUO Wei YANG Huan ZHANG Bing-bing. Effect of inhibition of heat shock protein Gp96 expression on alcoholic liver fibrosis in mice by CRISPR/Cas-9 technique[J]. Acta Anatomica Sinica. 2021, 52(5): 777-783 https://doi.org/10.16098/j.issn.0529-1356.2021.05.016

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