摘要
目的:探讨高迁移率族蛋白1-Toll样受体4(HMGB1-TLR4)信息途径在子痫前期(PE)内皮细胞屏障功能障碍过程中的作用。方法:原代培养C57BL/6小鼠胎盘滋养细胞(PMT)与腹主动脉内皮细胞(MAECs),PMT或HMGB1 siRNA干预后的PMT缺氧培养,将缺氧培养PMT上清或rHMGB1作用于MAECs。检测缺氧培养PMT的细胞活性、细胞凋亡、高迁移率族蛋白(HMGB1)基因与蛋白表达水平;检测缺氧培养PMT上清处理MAECs后,Toll样受体4(TLR4)蛋白表达及单层细胞通透性。再利用TLR4基因敲除小鼠(TLR4-/-)MAECs,观察缺氧培养PMT上清对TLR4基因敲除MAECs通透性的改变。结果:缺氧24h可诱导PMT HMGB1 mRNA、蛋白表达增加,缺氧培养上清HMGB1含量显著高于常氧培养组(P<0.05);PMT缺氧培养上清可明显增加MAECs单层细胞的通透性及TLR4蛋白表达,siRNA沉默PMT HMGB1或MAECs TLR4基因敲除可逆转PMT缺氧培养诱导的MAECs单层细胞通透性(P<0.05)。结论:缺氧小鼠滋养细胞释放HMGB1通过TLR4途径增加MAECs通透性。
Objective: To explore the role of high mobility group box 1( HMGB1)-Tolllike receptor 4( TLR4) pathway on the pathologic processes of endothelial monolayer permeability in preeclampsia. Methods: To culture the primary mouse trophoblast( PMT) and mouse aortic endothelial cells( MAECs). To cultured the PMT or PMT with HMGB1 silenced by HMGB1 siRNA in hypoxia condition. We used the condition-medium( CM) of hypoxia PMT or rHMGB1 to deal with MAECS. We assessed the cell viability,cell apoptosis,HMGB1 protein and gene expression,condition-medium HMGB1 of hypoxia PMT. We detected the permeability and TLR4 protein expression of MAECs which cultured with CM of hypoxia PMT. We also investigated the permeability of Toll-like receptor 4- /-( TLR4- /-) MAECs which cultured with CM of hypoxia PMT. Results: HMGB1 mRNA,protein expression and condition medium( CM) levels of 24-hour hypoxic PMT higherly increased as compared to those of normoxic PMT. The 24 hour hypoxic PMT CM increased the permeability of the MAECs monolayer and increasedthe Toll-like receptor 4( TLR4) protein expression of MAECs,which could be restored by PMT with HMGB1 siRNA transfection before hypoxia. MAECs with silenced TLR4 restored the 24 hour hypoxic PMT CM inducing the permeability of MAECs. Conclusions: Hypoxic PMT increased MAECs monolayer permeability via the HMGB1-TLR4 pathway,which could play an important role in the mechanism of urine protein in PE.
出处
《现代妇产科进展》
CSCD
2014年第1期13-17,共5页
Progress in Obstetrics and Gynecology
基金
国家自然基金青年基金(No:31101065)
