金匮肾气丸调控NLRP3/Caspase-1/IL-1β通路减轻SH-SY5Y细胞炎症和凋亡OA

Objective To explore the mechanism of Jingui Shenqi Pill(JSP)on 1-methyl-4-phenyl-pyridine(MPP+)-induced SH-SY5Y cells through NOD-like receptor protein 3(NLRP3)/cysteinyl aspartate specific proteinase(Caspase-1)/interleukin(IL)-1β signaling pathway in vitro.Methods SH-SY5Y cells were divided into Normal group,Model group and JSP(25,50,75,100 μg/mL)group.SH-SY5Y cells were induced by MPP+,cell viability was assessed using the cell counting kit-8(CCK-8)assay,and cytotoxicity was determined by measuring lactate dehydrogenase(LDH)release.The cell morphology of each group was observed by inverted microscope.ELISA was used to detect the interleukin(IL)-10,IL-1β,tumor necrosis factor-α(TNF-α),IL-6,and IL-18.Apoptosis was detected by flow cytometry.The protein expression levels of NLRP3,Caspase-1,B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),and C-Caspase-3 were detected by Western Blot.The mean fluorescence intensity of NLRP3,Caspase-1,and IL-1 β was detected using immunofluorescence.Results The optimal concentration of JSP was 50 μg/mL.Cells in the Model group predominantly exhibited a rounded morphology,with shortened or absent dendritic processes,poor adherence,and reduced cell number.Treatment with JSP ameliorated these morphological alterations:the number of rounded cells decreased,neurite outgrowth was more prominent,cell adherence improved,and cell number increased.Compared to the Normal group,the Model group demonstrated a significant decrease in SH-SY5Y cell viability,IL-10 levels,and Bcl-2 protein expression(P＜0.01).There was a significant increase in cytotoxicity,apoptosis rate,levels of TNF-α,IL-18,IL-6,IL-1β,and protein expression of Bax,C-Caspase-3,NLRP3,and Caspase-1(P＜0.05,P＜0.01).Compared to the Model group,the JSP group showed a significant increase in cell viability,IL-10 levels,and Bcl-2 protein expression(P＜0.05,P＜0.01),along with a significant decrease in cytotoxicity,apoptosis rate,levels of TNF-α,IL-18,IL-6,IL-1β,and protein expression of Bax,C-Caspase-3,NLRP3,and Caspase-1(P＜0.05,P＜0.01).Conclusions JSP exerts a protective effect against MPP+-induced damage in SH-SY5Y cells,with the most significant efficacy observed at a concentration of 50 μg/mL,markedly enhancing cell viability,improving morphology,and reducing cytotoxicity.Its mechanism may involve the inhibition of the NLRP3/Caspase-1/IL-1β signaling pathway,thereby alleviating inflammatory responses and suppressing apoptosis.