积雪草酸抑制TLR4/MyD88/NF-κB信号通路对高糖诱导的滋养层细胞焦亡的影响  

Effect of asiatic acid on high glucose induced trophoblast cell pyroptosis by inhibiting the TLR4/MyD88/NF-κB signaling pathway

作　　者：姚姝 张丽杰 YAO Shu;ZHANG Lijie(Production Department,Xi’an Gaoxin Hospital,Xi’an,Shaanxi 710000,China)

机构地区：[1]西安高新医院产二科,陕西西安710000

出　　处：《中国优生与遗传杂志》2024年第9期1826-1832,共7页Chinese Journal of Birth Health & Heredity

摘　　要：目的分析积雪草酸(AA)抑制Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核转录因子kappa B(NF-κB)信号通路对高糖诱导的滋养层细胞焦亡的影响。方法用25 mmol/L葡萄糖处理细胞,构建高糖HTR8/SVneo细胞模型。用脂多糖(LPS)、TAK242和不同浓度的AA分别处理HTR8/SVneo细胞。MTT法测定细胞活力,Transwell小室测定细胞侵袭,划痕实验测定细胞迁移,流式细胞仪测定凋亡,扫描电镜测定焦亡,免疫印迹法(Westernblot)测定TLR4/MyD88/NF-κB通路及焦亡蛋白表达。结果HG处理后HTR8/SVneo细胞凋亡率,IL-18、IL-1β水平,以及TLR4、MyD88、p-NF-κB、NLRP3、IL-1β、caspase1表达增加,细胞活力、侵袭数、迁移率减少(P<0.05);AA处理后细胞活力、侵袭数、迁移率增加,凋亡率,IL-18、IL-1β水平,以及TLR4、MyD88、p-NF-κB、NLRP3、IL-1β、caspase1表达减少(P<0.05);LPS处理后凋亡率,IL-18、IL-1β水平,以及TLR4、MyD88、p-NF-κB、NLRP3、IL-1β、caspase1表达增加,细胞活力、侵袭数、迁移率减少(P<0.05);TAK242处理后细胞活力、侵袭数、迁移率增加,凋亡率,IL-18、IL-1β水平,以及TLR4、MyD88、p-NF-κB、NLRP3、IL-1β、caspase1表达减少(P<0.05)。HTR8/SVneo组、TAK242组细胞形状和边界规则,HG组、LPS组细胞体积增大,微绒毛脱落,细胞膜肿胀形成多个泡状突起。结论AA可能抑制TLR4/MyD88/NF-κB通路,从而减弱高糖诱导的滋养层细胞焦亡。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-1βlevels and the expression of TLR4,MyD88,p-NF-κB,NLRP3,IL-1β,and caspase1 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 apoptosis rate,IL-18,and IL-1βlevels 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-1βlevels 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-1βlevels 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 T

关 键 词：积雪草酸 Toll样受体4/髓样分化因子88/核转录因子kappa B通路 滋养层 细胞焦亡 妊娠糖尿病 

分 类 号：R714.256[医药卫生—妇产科学]