白头翁皂苷A下调LRP1抑制滑膜炎性细胞的增殖、迁移和侵袭OA

Objective To investigate the effects and mechanisms of pulchinenoside A(PSA)on tumor necrosis factor-α(TNF-α)-induced abnormal proliferation,migration,and invasion of synovial fibroblasts.Methods A sy-novitis model was established by stimulating SW982 synovial cells with TNF-α.The low-density lipoprotein receptor-related protein 1(LRP1)was knocked down using LRP1-specific siRNA or inhibited by the receptor-associated protein(RAP).Cells were treated with various concentrations of PSA,and cell viability was assessed by CCK-8 assay.Protein levels of LRP1,matrix metalloproteinase-3(MMP3),and N-cadherin were measured by Western blotting.Proliferation was evaluated using an EdU incorporation assay.Migration and invasion capacities were examined by Transwell and wound-healing assays.mRNA expression of LRP1,MMP3,N-cadherin,IL-1β,IL-6,and TNF-α was detected by reverse transcription quantitative real-time PCR(RT-qPCR).Immunofluores-cence staining was used to visualize the expression of MMP3 and N-cadherin.The involvement of LRP1 was further verified with the inhibitor RAP.Results Compared with the TNF-α-treated model group,PSA treatment signifi-cantly reduced SW982 cell viability(P<0.05),downregulated MMP3 and N-cadherin expression(both P<0.01),and decreased the proportion of EdU-positive cells(P<0.05).Moreover,TNF-α stimulation upregulated LRP1 ex-pression compared with normal controls(P<0.01),whereas PSA treatment reversed this increase(P<0.05).LRP1 knockdown or inhibition markedly decreased MMP3 and N-cadherin levels compared with the model group(both P<0.01).Similarly,the combination of PSA and RAP led to significant reductions in MMP3 and N-cadherin expression compared with the model group(both P<0.01).Conclusion PSA inhibits TNF-α-induced abnormal proliferation,migration,and invasion of synovial,at least in part,by downregulating LRP1 expression.