Objective:This paper aims to explore the mechanism of Qianlie-Biertong suppository(QS)in improving prostatitis in vitro.Methods:The intestine absorbed drug solution of QS(IQS)was prepared ex vivo and characterized by UPLC-QE-Orbitrap-MS.The lipopolysaccharide(LPS)-induced PC-3 cell inflammation model of human prostate cancer cell lines was established.Cell viability was determined by MTT assay.The gene expression levels of inflammatory mediators were detected by qRT-PCR.The ex-pression levels of the key proteins involved in the Toll-like receptor 4(TLR4)signaling pathway were determined by Western blot.The nuclear translocation of nuclear factor κB/p65,activated protein-1(AP-1)subunit c-Jun,and interferon regulatory factor 3(IRF3)was detected by immunofluorescence.Results:Various bioactive components including berberine,chlorogenic acid,and fer-ulic acid were identified in IQS.IQS treatment at concentrations of 6.25 to 400 μg/mL had no significant effect on the cell viability of PC-3 cells.The gene expression levels of inflammatory mediators under LPS exposure,including TNFA,IL-6,CXCL10,MCPl,and COX2 were markedly inhibited by IQS.IQS could downregulate the expression levels of phosphorylated AKT,IκBα,IKKα/β,p65,p38,c-Jun,TBK1,and IRF3 in a dose-dependent manner,which were key proteins of the TLR4 signaling pathway.Furthermore,IQS remarkably suppressed the nuclear translocation of p65,c-Jun,and IRF3 induced by LPS.Conclusion:IQS mitigated LPS-induced in-flammatory response in PC-3 cells,which may be associated with the inhibitory effects of IQS on the TLR4 signaling pathway.
Qianlie-biertong suppositoryIntestine absorbed drug solutionUPLC-QE-Orbitrap-MS technologyPC-3 cellsLi-popolysaccharideInflammatory responseProstatitisTLR4 signaling pathway
维普
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