Effect of asiatic acid on high glucose induced trophoblast cell pyroptosis by inhibiting the TLR4/MyD88/NF-κB signaling pathway
Objective This study aims to analyze the effect of asiatica acid(AA)on high glucose induced trophoblast cell pyroptosis by inhibiting the Toll like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor kappa B(NF-κB)signaling pathway.Methods The high-glucose HTR8/SVneo cell model was constructed by treating the cells with 25 mmol/L glucose.HTR8/SVneo cells were treated with LPS,TAK242 and AA at different concentrations.MTT method was applied to measure cell viability.Transwell chamber was used to measure cell invasion.Scratch experiment was applied to determine cell migration.Flow cytometry was applied to measure apoptosis.Scanning electron microscopy was applied to determine pyroptosis.Immunoblotting was applied to determine the TLR4/MyD88/NF-κB pathway and pyroptosis protein expression.Results After HG treatment,the apoptosis rate,IL-18,and IL-1β levels and the expression of TLR4,MyD88,p-NF-κB,NLRP3,IL-1β,and caspase 1 of HTR8/SVneo cells increased,while cell viability,invasion number,and migration rate decreased(P＜0.05).After AA treatment,the cell viability,invasion number,and migration rate increased,while the apop-tosis rate,IL-18,and IL-1β levels and the expression of TLR4,MyD88,p-NF-κB,NLRP3,IL-1β,and caspase1 decreased(P＜0.05).After LPS treatment,the apoptosis rate,IL-18,and IL-1β levels and the expression of TLR4,MyD88,p-NF-κB,NLRP3,IL-1β,and caspase1 increased,while cell viability,invasion number,and migration rate decreased(P＜0.05).After TAK242 treatment,the cell viability,invasion number,and migration rate increased,while the apoptosis rate,IL-18,and IL-1 βlevels and the expression of TLR4,MyD88,p-NF-κB,NLRP3,IL-1β,and caspase1 decreased(P＜0.05).The cell shape and boundaries in the HTR8/SVneo group and TAK242 group were regular,while in the HG group and LPS group,the cell volume increased,microvilli shed,and the cell membrane swelled to form multiple vesicular protrusions.The cell morphology in the L-AA group,M-AA group,and H-AA group improved.Conclusion AA may inhibit the TLR4/MyD88/NF-κB pathway,thereby weakening high glucose induced trophoblast cell pyroptosis.
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