人滋养细胞TLR3信号通路活化抑制血管生成因子表达

摘要

目的 研究人类妊娠早期滋养细胞Toll样受体3(TLR3)活化对胎盘血管生成相关因子表达的影响,探讨该通路在妊娠期高血压疾病中的作用.方法 以TLR3配体刺激原代滋养细胞和永生化滋养细胞系swan71,不同时间点收集上清液及细胞.ELISA测定培养上清液中sFlt-1和PIGF浓度,real-time PCR法测定上述分子mRNA表达水平.结果 Poly(I∶C)刺激swan71后24、48和120 h,sFlt-1浓度显著高于未处理组(P＜0.05).Poly(I∶C)刺激原代滋养细胞后sFlt-1 mRNA水平升高,PlGF mRNA水平下降(P＜0.05).Poly(I∶C)诱导sFlt-1 mRNA的表达呈时间和剂量依赖性,24 h时效分析见其在处理2h达到峰值,PlGF mRNA则跌至最低水平(P＜0.05).Poly(I∶C)处理8～12 h,TLR3 mRNA水平亦显著升高(P＜0.05).结论 滋养细胞TLR3信号通路激活诱导sFlt-1表达,抑制PlGF表达,导致血管生成障碍,可能参与妊娠期高血压疾病的发生.%Objective To investigate the effect of TLR3 ligation on expression of angiogenic factors in primary trophoblast cells and to evaluate the role of TLR3 signal pathway in the pathogenesis of pregnant hypertension.Methods The first trimester human trophoblast cells and immortalized human trophoblast cell line Swan71 were stimulated with Poly(Ⅰ ∶ C) (TLR3 specific ligand).sFlt-1 and PlGF protein and mRNA expression were evaluated by ELISA and Real-time PCR respectively at different time points.TLR3 mRNA expression was also determined by Real-time PCR.Results 24,48 and 120 h after Poly(I:C) stimulation,concentration of sFlt-1 in the conditioned media of Swan71 was significantly increased as compared with non-treatment control (P ＜ 0.05).Poly (I∶ C) (10 μg/mL) significantly induced sFlt-1 mRNA (P ＜ 0.05),while suppressed PlGF mRNA (P ＜ 0.05) in primary trophoblast cells.Induced expression of sFlt-1 mRNA showed a time and dose-dependent manner,which reached the highest effect 2 h after Poly (I ∶ C) stimulation (P ＜ 0.05).At the same time,PlGF mRNA expression was decreased significantly (P ＜ 0.05).Expression of TLR3 mRNA was significantly increased 8 to 12 h after Poly (I∶C) stimulation(P ＜ 0.05).Conclusions TLR3 activation induces synthesis and secretion of sFlt-1,while inhibits PlGF in trophoblast cells,thus lead to disturbance of angiogenesis and be involved in the pathophysiology of hypertensive disorders in pregnancy.