茵陈蒿汤调节巨噬细胞活化抗胆汁淤积性肝损伤的作用机制OA

Objective:This study aims to investigate the mechanism of Yinchenhao Tang(YCHT)in regulating macrophage polarization to alleviate cholestatic liver injury,focusing on the TLR4/NF-κB signaling pathway as the entry point.Methods:Cholestasis was induced in Wistar rats through a single gavage of 100 mg·kg-1 α-naphthyl isothiocyanate(ANIT)dissolved in olive oil.The animals were randomly divided into four groups:Model group,YCHT group,ursodeoxycholic acid(UDCA)group(n=10),and a blank group(n=10)that received only 5 mL·kg-1 olive oil.The YCHT group received 9.23 g·kg-1·day-1 of YCHT by gavage,and the UDCA group was treated with 0.1 g·kg-1·day-1 of UDCA suspension.Both the normal and model groups were given an equal volume of normal saline,all for three consecutive days.Serum liver function was assessed using an automatic biochemical analyzer.Hematoxylin-eosin(HE)staining was used to observe liver tissue morphology.Levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),transforming growth factor-β(TGF-β),and interleukin-10(IL-10)were quantified in liver homogenate supernatants via enzyme-linked immunosorbent assay(ELISA).Western blot analysis measured the relative protein expression of Toll-like receptor 4(TLR4),nuclear factor-κB(NF-κB),CD206,inducible nitric oxide synthase(iNOS),CD86,and arginase-1(Arg-1).The relative mRNA expression of TLR4/NF-κB,CD206,iNOS,CD86,and Arg-1 in liver tissue was evaluated using real-time quantitative PCR.Results:Compared with the normal group,the model group exhibited significantly elevated levels of alkaline phosphatase(ALP),total bile acid(TBA),total bilirubin(TBil),aspartate aminotransferase(AST),and alanine aminotransferase(ALT)(P<0.01).There was a portal area expansion and pronounced inflammatory cell infiltration.The expression of pro-inflammatory markers TNF-α and IL-1β was significantly upregulated(P<0.01),and macrophage markers CD86 and CD206 showed positive expression.Protein and mRNA expressions of iNOS and CD86 were significantly elevated(P<0.01).The mRNA and protein expressions of the related pathway molecules TLR4 and NF-κB were significantly increased(P<0.01).Compared with those in the model group,the liver function indicators in the YCHT group showed significant decreases(P<0.05,P<0.01).The bile duct hyperplasia was significantly alleviated,and the tissue structure became more orderly.The levels of IL-1β and TNF-α were significantly reduced(P<0.01),while the expression levels of IL-10 and TGF-β significantly increased(P<0.05,P<0.01).The expression of CD86 significantly decreased(P<0.01),and the expression of CD206 significantly increased(P<0.01).The protein and mRNA expressions of iNOS and CD86 significantly decreased(P<0.01),and those of Arg-1 significantly increased(P<0.01).The protein and mRNA expressions of CD206 significantly increased(P<0.05,P<0.01),and the mRNA and protein expressions of related pathway molecules TLR4 and NF-κB significantly decreased(P<0.01).Conclusion:YCHT ameliorates cholestatic liver injury in rats by improving bile metabolism,reducing bile duct dilatation,and mitigating inflammation.These effects are achieved through the inhibition of M1 macrophage activation and the promotion of M2 macrophage polarization,likely via modulation of the TLR4/NF-κB signaling pathway.