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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 investigate the role of high mobility group box 1 (Toll-like receptor 4) (HMGB1-TLR4) signaling pathway in the process of endothelial dysfunction in endothelial cells of preeclampsia (PE). Methods: Primary hypoxic culture of PMT after C57BL / 6 mouse placental trophoblast cells (PMT) and abdominal aorta endothelial cells (MAECs), PMT or HMGB1 siRNA was performed. Hypoxic culture of PMT supernatant or rHMGB1 was performed on MAECs . The cell viability, apoptosis and HMGB1 gene and protein expression of hypoxic cultured PMT were detected. The expression of Toll-like receptor 4 (TLR4) and the expression of TLR4 protein Layer cell permeability. The TLR4 gene knockout mice (TLR4 - / -) MAECs were used to observe the changes of the permeability of TLR4 knockdown MAECs by hypoxic cultured PMT supernatant. Results: HMGB1 mRNA was induced by hypoxia for 24 h, the protein expression of HMGB1 was increased, and the content of HMGB1 in hypoxic culture supernatant was significantly higher than that in normoxic group (P <0.05). The hypoxic culture supernatant of PMT significantly increased the expression of HMGB1 mRNA Permeability and TLR4 protein expression, siRNA silencing PMT HMGB1 or MAECs TLR4 knockout can reverse the monolayer permeability of MAECs induced by PMT hypoxia (P <0.05). CONCLUSION: The release of HMGB1 by trophoblast cells in hypoxic mice increases the permeability of MAECs through the TLR4 pathway.