Honokiol Regulates FXR-LXRs Signaling Pathway To Improve The Process Of Liver Fibrosis

Objective:Honokiol(Ho)is one of the main chemical components of the traditional Chinese medicine magnolia officinalis,which has good antibacterial and anti-inflammatory effects.In this study,we devoted to investigating the control and the potential molecular machines of Honokiol on liver fibrosis based on transforming growth factor TGF-β-stimulated HSCs in vitro and intraperitoneal injection of thioacetamide(TAA)in vivo.Methods:After stimulation with TGF-β,HSCs cells were treated with a dose of 0-50μM of Ho for 24 h.The influence of Ho on the survival rate of HSCs cells was observed by MTT experiment.The gradient concentration(0,1.56,3.125,6.25,6.25,12.5,50 μM)and Ho were incubated with HSCs for 24 h,total protein and RNA were extracted,and protein and gene expression levels were tracked by Western blot and RT-PCR,and immunofluorescence analysis was also used to further verification.The C57BL/6 male mice were randomly divided into six groups:normal group,TAA group,and the administration group.Different doses of Ho(10 or 20mg/kg)were given daily,and the positive control.The group was given curcumin(Curcumin,Cur,20 mg/kg),and the group given alone and Ho(20 mg/kg).Four weeks of liver fibrosis modeling experiments were performed.Except the normal group,mice were sacrificed 6 days after the last administration,and blood samples and livers were collected.ALT and AST levels in serum were detected using the kit method.Paraffin sections of the extracted liver tissues were used to observe the pathological changes of mouse liver tissues by H&E,Sirius red staining,and immunohistochemical staining.At the same time,Western blotting,RT-PCR and immunofluorescence staining were used to detect the expression of α-SMA,FXR,LXRs,P2X7r,NLRP3,Caspase-1,IL-1β and other proteins and mRNA.Results:The MTT results showed that Ho had no effect on the growth of HSCs cells in the range of 0～12.5μM.Ho could effectively inhibit the protein and fluorescent expression of α-SMA and Collagen-I.The activation of FXR and LXRs inhibited the release of various inflammatory factors such as P2X7r and NLRP3.The administration of Ho effectively inhibited the increase of serum ALT and AST levels in mice induced by TAA and reduced the expression of protein and mRNA of α-SMA and Collagen-I.And Ho activated the expression of FXR and LXRs to inhibit the release of various inflammatory factors such as P2X7r,NLRP3,and IL-1β.Conclusion:Ho can down-regulate the expression of liver fibrosis markers α-SMA and Collagen-I,and actually inhibited the release of pro-inflammatory factors,which may improve liver fibrosis by regulating FXR-LXRs-related signaling pathways.