摘要
目的 研究内质网应激(ERS)在体外缺氧诱导子痫前期(PE)滋养细胞模型中调控凋亡的作用及机制.方法 收集PE胎盘和健康胎盘,检测细胞凋亡率及ERS标志基因:葡萄糖调节蛋白 78(GRP78)、ERS凋亡基因C/EBP同源蛋白(CHOP)、磷酸化JNK(p-JNK)、含半胱氨酸的天冬氨酸蛋白水解酶-12(Cleaved caspase-12)的表达水平.培养胎盘滋养细胞HTR8/SVneo,采用低氧(1%O2)处理 24 h诱导PE细胞模型,采用ERS激动剂处理细胞,在PE细胞模型诱导过程中转染CHOP siRNA或给予JNK拮抗剂、Caspase-12 抑制剂,检测细胞增殖水平(D值)、凋亡率及GRP78、CHOP、p-JNK、Cleaved caspase-12 的表达水平.结果 PE胎盘细胞凋亡率及GRP78、CHOP、p-JNK、Cleaved caspase-12 的表达水平高于健康胎盘(P＜0.05),且细胞凋亡率与 GRP78、CHOP、p-JNK、Cleaved caspase-12 的表达水平呈正相关;PE组及ERS激动剂组细胞的D值低于对照组,凋亡率及GRP78、CHOP、p-JNK、Cleaved caspase-12 的表达水平高于对照组(P＜0.05);缺氧诱导PE模型过程中转染CHOP siRNA降低CHOP表达,给予JNK拮抗剂降低p-JNK表达,给予Caspase-12 抑制剂降低Cleaved caspase-12 表达后,细胞D值升高,凋亡率及GRP78 的表达水平降低(P＜0.05).结论 缺氧诱导PE滋养细胞模型中ERS,通过CHOP、JNK和Caspase-12 介导细胞凋亡.
Abstract
Objective To study the role and mechanism of endoplasmic reticulum stress(ERS)in regulating the apoptosis of hypoxia-induced preeclampsia(PE)trophoblasts in vitro.Methods PE placenta and healthy placenta were collected,and then the apoptosis rate and the levels of ERS marker gene glucose regulated protein 78(GRP78),ERS apoptosis gene C/EBP homologous protein(CHOP),phosphorylated JNK(p-JNK)and Cleaved caspase-12 were examined.The placental trophoblasts HTR8/SVneo were cultured.A PE cell model was induced by exposing to low oxygen(1%O2)for 24 h.The cells were treated with ERS agonists.During the induction of the PE cell model,the cells were transfected with CHOP siRNA or treated with JNK inhibitor and Caspase-12 inhibitor.Cell proliferation(D value),apoptosis rate and the levels of GRP78,CHOP,p-JNK and Cleaved caspase-12 were measured.Results The apoptosis rate and the levels of GRP78,CHOP,p-JNK and Cleaved caspase-12 in PE placenta were higher than those in healthy placenta(P＜0.05).Cell apoptosis rate was positively correlated with the levels of GRP78,CHOP,p-JNK,and Cleaved caspase-12.The D value of cells in the PE group and ERS agonist group were lower than those in the control group,and the apoptosis rate and the the levels of GRP78,CHOP,p-JNK and Cleaved caspase-12 were higher than those in the control group(P＜0.05).Transfection of CHOP siRNA reduced CHOP expression,JNK inhibitor reduced p-JNK expression,and Caspase-12 inhibitor reduced Cleaved caspase-12 expression in the hypoxia-induced PE model,leading to increased D value,reduced apoptosis rate,and lowered GRP78 expression(P＜0.05).Conclusions ERS mediates cell apoptosis through CHOP,JNK,and Caspase-12 in the hypoxia-induced PE trophoblast model.
NETL
NSTL
维普
万方数据