| Aims:Pyroptosis is a primary event involved in liver fibrosis,but the mechanisms of pyroptosis have not been clarified.Calcium-binding protein HRC is involved in liver fibrosis,but its role in hepatocyte pyroptosis has not been explored.We investigated the role of HRC in hepatocyte pyroptosis during liver fibrosis.Methods:The location and expression of HRC was detected by immunohistochemistry in the liver tissues of patients with liver fibrosis and CCl4-induced model.To investigate the role of HRC in hepatocyte,we established L02 cells with HRC overexpression and then detected expression of NLRP3,caspase-1 and GSDMD by RT-q PCR and Western blot.The number of Caspase-1/PI double-positive hepatocyte was estimated by flow cytometry.LDH and HMGB1 in supernatant of L02 cells with HRC overexpression were detected by LDH assay and ELISA assay.We collected conditioned medium from L02 cells to treat LX-2cells.LX-2 cells incubated in conditioned medium was assessed by RT-q PCR and Western blot for markers of heapatic stellate cells activation,proliferation and migration.Meanwhile,proliferation of LX-2 was examined by CCK-8 and Edu assay,migration was examined by Transwell assay.Result:Expression of HRC was higher in patients with liver fibrosis and CCl4-induced model.HRC promoted expression of NLRP3,caspase-1,GSDMD and HMGB1 at protein level,but not m RNA level excepted NLRP3.L02 cells with HRC overexpression showed markedly hepatocyte pyroptotic cell death,with more than a 3-fold increase in active caspase-1/PI double-positive cells.In addition,LDH and HMGB1 levels in supernatant of L02 with HRC overexpreesion were significantly elevated compared with control.Conditioned medium from pyroptotic hepatocytes promoted activation,proliferation and migration of hepatic stellate cells.Conclusion:Our study demonstrates that HRC plays an important role in hepatocyte pyroptosis and release of HMGB1,which promotes activation,proliferation and migration of HSCs during liver fibrosis. |
PDF Full Text Request