Role of growth factor receptor-bound 2 in CCl4-induced hepatic fibrosis

Background: Growth Factor Receptor-bound 2 (GRB2) plays a crucial role in regulation of cellular function including proliferation and differentiation, and we previously identified GRB2 as promoting HSCs (HSCs) proliferation. However, the underlying mechanisms that are involving in the regulation of GRB2 in hepatic fibrogenesis remain unknown. Methods: In the present study, we tested the function of GRB2 in hepatic fibrosis. Hepatic fibrosis was induced by subcutaneous CCl4 administration at a dose of 3 mL/kg in rats. The rat HSC cell line HSC-T6 were cultured for proliferation investigation by CCK-8 and BrdU incorporation method. The levels of GRB2, HMGB1, PI3K/AKT, COL1A1 and alpha-SMA were analyzed by western blot or real-time PCR. Results: showed that the expression of GRB2 and HMGB1 was obviously increased in liver tissues of hepatic fibrosis rats accompanied by up-regulation of COL1A1 and alpha-SMA. In cultured HSCs, application of exogenous HMGB1 induced cell proliferation and cell proliferation rate concomitantly with up-regulation of GRB2 expression and PI3K/AKT phosphorylation. The effects of HMGB1-induced proliferation of HSCs and up-regulation of COL1A1 and alpha-SMA were abolished by GRB2 siRNA. HMGB1-induced proliferation of HSCs and up-regulation of COL1A1 and a-SMA was reversed in the presence of LY294002, an inhibitor of PI3K inhibitor. Conclusions: These findings suggest that GRB2 plays an important role in CCl4-induced hepatic fibrosis by regulating HSCs' function, and up-regulation of GRB2 induced by HMGB1 is mediated via the PI3K/AKT pathway. (C) 2017 Published by Elsevier Masson SAS.