The Role and Molecular Regulatory Mechanism of A Disintegrin and Metalloprotease 9 in Alcoholic Liver Fibrosis

Abstract

A disintegrin and metalloprotease 9 has been reported to play a key role in many diseases. However, the underlying effects and mechanisms of a disintegrin and metalloprotease 9 in alcoholic liver fibrosis have not been well understood. To investigate the role of a disintegrin and metalloprotease 9 and its regulatory molecular mechanism in alcoholic liver fibrosis, a single guide ribonucleic acid targeting the a disintegrin and metalloprotease 9 sequence of mice was adopted by clustered regularly interspaced short palindromic repeats/Cas9 technology to inhibit its expression in this study. Subsequently, hepatic stellate cell-T6 and male mice C57BL/6J were respectively divided into 4 groups and each group was treated differently. In vivo experiment, the normal group was fed with control Lieber-DeCarli TP4030C; the saline+alcohol group was injected with saline; the A disintegrin and metalloprotease 9-single guide ribonucleic acid+alcohol group was injected with effective single guide ribonucleic acid plasmids and the c-Jun N-terminal kinase inhibitor+alcohol group was injected with c-Jun N-terminal kinase inhibitor SP600125. All mice except for the normal group received Lieber-DeCarli TP4030A and carbon tetrachloride. Furthermore, examination of serum aspartate transaminase and alanine transaminase activities and the pathological analysis by hematoxylin-eosin staining, picric acid-sirius red staining, Hoechst 33 258 staining confirmed the pro-fibro genic effect of a disintegrin and metalloprotease 9. Compared with the saline+alcohol group, the levels and the expression of a disintegrin and metalloprotease 9, p-c-Jun, cytochrome P450 2E1, tumor necrosis factor alpha, alpha-smooth-muscle actin and Bcl-2-associated X in the A disintegrin and metalloprotease 9- single guide ribonucleic acid+alcohol group and the c-Jun N-terminal kinase inhibitor+alcohol group decreased significantly (p<0.05 or p<0.01), while the expression of heat shock protein 70, proliferating cell nuclear antigen and vascular endothelial growth factor increased significantly (p<0.05 or p<0.01). In addition, the disintegrin and metalloprotease 9-single guide ribonucleic acid+alcohol group and the c-Jun N-terminal kinase inhibitor+alcohol group in vitro experiment could inhibit the proliferation of hepatic stellate cells and promote apoptosis. Collectively, our results indicated that a disintegrin and metalloprotease 9 could promote alcoholic liver fibrosis in mice by activating the c-Jun N-terminal kinase signaling pathway, which is expected to make a breakthrough in the gene therapy of alcoholic liver fibrosis.