茵陈蒿汤调控细胞焦亡干预胆汁淤积性肝损伤的机制OA

Objective:To explore the mechanism by which Yinchenhao Tang intervenes in α-naphthylisothiocyanate(ANIT)-induced cholestatic liver injury by regulating the Takeda G-protein-coupled receptor 5(TGR5)/NOD-like receptor protein 3(NLRP3)/cysteine aspartate-specific protease-1(Caspase-1)pyroptosis signaling pathway.Methods:Forty male Wistar rats were randomly assigned into blank,model,ursodeoxycholic acid,and Yinchenhao Tang groups.Except the blank group,other groups were treated with ANIT dissolved in olive oil for the modeling of cholestatic liver injury.Ursodeoxycholic acid(0.1 g·kg-1)and Yinchenhao Tang(9.23 g·kg-1)were administered by gavage.The blank group and the model group were administrated with the same amount of pure water,once a day for 3 days.The blood and liver tissue samples were collected,and the serum levels of liver function indicators were measured by an automatic biochemical analyzer.Hematoxylin-eosin staining was employed to observe the pathological changes of the liver.The levels of interleukin(IL)-1β and IL-18 in the liver tissue were determined by ELISA.The mRNA levels of IL-1β,IL-18,TGR5,NLRP3,apoptosis-associated speck-like protein containing a CARD(ASC),Caspase-1,and GSDMD in the liver tissue were assessed by Real-time PCR.The protein levels of TGR5,NLRP3,ASC,Caspase-1,and GSDMD in the liver tissue were determined by Western blot.Results:Compared with the blank group,the model group showed elevated levels of alanine amino-transferase(ALT),aspartate transferase(AST),alkaline phosphatase(ALP),total bile acid(TBA),and total bilirubin(TBil)in the serum(P<0.01),inflammatory cell infiltration,hepatocyte swelling,and bile duct epithelial cell proliferation in the liver,raised levels of IL-1β and IL-18 in the liver tissue(P<0.01),down-regulated mRNA and protein levels of TGR5(P<0.01),up-regulated mRNA levels of IL-18(P<0.01),ASC(P<0.01),Caspase-1(P<0.01),GSDMD(P<0.01),IL-1β(P<0.05),and NLRP3(P<0.05),and up-regulated protein levels of NLRP3(P<0.01),ASC(P<0.01),Caspase-1(P<0.01),and GSDMD(P<0.05).Compared with the model group,the ursodeoxycholic acid group showed declined levels of AST(P<0.01),TBA(P<0.01),TBil(P<0.01),and ALT(P<0.05)in the serum,lowered levels of IL-1β and IL-18 in the liver tissue(P<0.01),down-regulated mRNA levels of NLRP3(P<0.01),Caspase-1(P<0.01),GSDMD(P<0.01),IL-1β(P<0.05),IL-18(P<0.05),and ASC(P<0.05),up-regulated mRNA and protein levels of TGR5(P<0.05),and down-regulated protein levels of NLRP3,ASC,Caspase-1,and GSDMD(P<0.05).Compared with the model group,the Yinchenhao Tang group showed lowered levels of ALT,AST,ALP,TBA,and TBil in the serum(P<0.01),declined levels of IL-1β and IL-18 in the liver tissue(P<0.01),down-regulated mRNA levels of IL-1β(P<0.01),NLRP3(P<0.01),ASC(P<0.01),Caspase-1(P<0.01),GSDMD(P<0.01),and IL-18(P<0.05),up-regulated mRNA and protein levels of TGR5(P<0.01),and down-regulated protein levels of Caspase-1 and GSDMD(P<0.05).The liver tissue of the administration groups showed reduced infiltration of inflammatory cells,reduced swelling of hepatocytes,and alleviated proliferation of bile duct epithelial cells.Conclusion:Yinchenhao Tang can ameliorate ANIT-induced cholestatic liver injury by regulating the hepatocyte pyroptosis mediated by the TGR5/NLRP3/Caspase-1 signaling pathway.