红景天苷抑制肝细胞焦亡改善肝纤维化的作用机制OA

Objective Salidroside(SAL),the main active component of Rhodiola rosea L.,was investigated in this study to deeply analyze the mechanism by which SAL improved hepatic fibrosis by inhibiting hepatocyte pyroptosis.Methods AML-12 cells were induced with lipopolysaccharide(LPS)combined with adenosine triphosphate(ATP)for 4 h,followed by treatment with different concentrations of SAL for 24 h.The protein expressions of NLRP3,ASC,cleaved caspase 1 and GSDMD were detected by Western blot.The mRNA levels of Nlrp3,Casp1 and Il1 b in cells were determined by RT-qPCR.The levels of inflammatory factors IL-6 and IL-1β in hepatocyte supernatants were analyzed by ELISA.Mouse primary hepatic stellate cells(HSCs)were treated with the conditioned medium from mouse primary hepatocytes stimulated by LPS+ATP.After SAL administration,the protein expressions of α-SMA,P2X7R,NLRP3,cleaved caspase 1,IL-23 and mature-IL-1β were detected by Western blot.Results Compared with normal group,LPS+ATP successfully induced AML-12 cell injury and established a hepatocyte pyroptosis model.Western blot results showed that SAL treatment significantly reduced the protein expressions of NLRP3,ASC,cleaved caspase 1 and GSDMD(P<0.01,P<0.001).RT-qPCR results indicated that SAL significantly decreased the mRNA levels of Nlrp3,Casp1 and Il1 b in AML-12 cells(P<0.001).ELISA results demonstrated that SAL could significantly inhibit the extracellular release of IL-1β and IL-6(P<0.001).The experiment also found that the conditioned medium from mouse primary hepatocytes stimulated by LPS+ATP could successfully induce the activation of mouse primary HSCs,and SAL treatment significantly reduced the protein expressions of α-SMA,P2X7R,NLRP3,cleaved caspase 1,IL-23 and mature-IL-1β in HSCs(P<0.001).Conclusion SAL can effectively downregulate the expression of inflammatory factors in hepatocytes,inhibit cell pyroptosis,and regulate the interaction between hepatocytes and hepatic stellate cells.These results suggest that SAL may serve as a potential candidate drug for anti-hepatic fibrosis.