Effect of S100A6 on activation of hepatic stellate cells in mice

Abstract

Objective To determine the expression of S100A6 in liver tissues and investigate its role in the activation of hepatic stellate cells (HSCs). Methods Twelve C57BL/6 mice were randomly assigned to control group and liver fibrosis group. The mice in the latter group was administrated with carbon tetrachloride for 6 weeks to induce liver fibrosis. The liver tissue was harvested for Masson staining and S100A6 detection by immunohistochemistry. The expression of S100A6 at mRNA and protein levels was detected by RT-qPCR and Western blotting. After primary HSCs were isolated and cultured, their morphological changes in quiescence and activation were observed, and the mRNA and protein levels of S100A6 were detected after cultured for different time periods. Additionally, siRNA was utilized to interfere S100A6 expression in HSCs, and the mRNA and protein levels of S100A6, Collagen Ⅰ and α-SMA were tested by RT-qPCR and Western blotting. Results Masson staining displayed fibrosis in liver tissue in the mice of the liver fibrosis group, and S100A6 was also significantly expressed in the corresponding fibrous cord area of liver tissue in liver fibrosis group, and its mRNA and protein levels were obviously enhanced in the liver tissue when compared to the control group (P < 0.05). In vitro, HSCs auto-activated with the process of culture, and the mRNA and protein expression levels of S100A6 were gradually increased in a time depended manner in the HSCs (P < 0.05 or P < 0.001). Undoubtedly, the interference of S100A6 by siRNA efficiency dramatically inhibited the mRNA and protein expression of S100A6, Collagen Ⅰ, α-SMA (P < 0.05 or P < 0.001). Conclusion S100A6 is specifically expressed in hepatic fibrosis and activated HSCs, and is an important protein regulating the activation of HSCs. Thus, S100A6 might play important role in the pathogenesis of liver fibrosis.

Keywords

• s100a6
• hepatic stellate cells
• liver fibrosis