Il-22 Inhibited The Proliferation And Activation Of Hepatic Stellate Cells Through The NLRP3/Caspase1/IL-1βsignaling Pathway

Objective:To explore whether interleukin-22(IL-22)can inhibit the proliferation of hepatic stellate cells by inhibiting the NLRP3/caspase1/IL-1β pathway,and to find a new target for the prevention and treatment of hepatic fibrosis caused by hepatic stellate cells.Methods:1.Treat hepatic stellate cells LX-2 with 5ng/m L transforming growth factor-β(TGF-β)for 24 h,induce the proliferation of hepatic stellate cells in vitro,and then use low medium and high doses(250pg/m L,500pg/m L,750pg/m L)IL-22 interfered with LX-2 cells induced by TGF-β for 48h;CCK8 experiment and flow cytometry analysis were used to detect the changes of LX-2 cell proliferation and apoptosis,Compare the proliferation and apoptosis of hepatic stellate cells before and after IL-22 treatment;2.First induce hepatic stellate cells with 5ng/m L TGF-β for 24 h,and then intervene with 750pg/m L IL-22 to induce TGF-β After 48 h,the LX-2 cells were treated with a 40μmol/L NOD-like receptor protein 3(NLRP3)agonist for 30 min,and then Western blot and RT-q PCR were used to detect the hepatic stellate cell α-Smooth muscle actin(α-SMA),type I collagen α1 chain gene(COL1A1)protein and m RNA expression changes,and detect NLRP3,cysteinyl aspartate specific proteinase1(caspase1)and interleuki-1β(IL-1β)in protein levels.Results:1.Compared with the blank group,after TGF-β induces LX-2,the proliferation of LX-2 cells increases and apoptosis decreases;after the intervention of low medium and high doses of IL-22,the proliferation of LX-2 cells decreases(P<0.05)),its apoptosis showed an upward trend with the increase of concentration,and the difference was statistically significant(P<0.05).2.Compared with the blank group,after TGF-β induced LX-2 cells,the protein and m RNA expression of α-SMA and COL1A1 in the cells showed an upward trend,and the protein expression levels of NLRP3,caspase1,and IL-1β increased(P<0.05);Compared with the TGF-βinduction group,after the intervention of IL-22,the expression of α-SMA,COL1A1,NLRP3,caspase1,and IL-1β in LX-2 cells decreased(P<0.05),while in the NLRP3 agonist group,The expression of the above indicators was reversed.Conclusions:IL-22 can inhibit the proliferation and activation of LX-2 by TGF-β,which may be achieved by inhibiting the NLRP3/caspase1/IL-1β signaling pathway.