目的 研究内质网应激(endoplasmic reticulum stress,ERS)标识物葡萄糖调节蛋白(glucose regulated protein,GRP)和CCAAT增强子结合蛋白同源蛋白(CCAAT/enhancer binding protein homologous protein,CHOP)在人胚肺成纤维细胞凋亡过程中的表达变化,初步探讨毒胡萝卜素对人胚肺成纤维细胞凋亡的作用及部分机制.方法 体外用毒胡萝卜素(TG)诱导人胚肺成纤维细胞(MRC-5)后,采用流式细胞仪检测细胞的凋亡变化,RT-PCR检测细胞中GRP78、CHOP mRNA的表达变化,Western blot检测GRP、CHOP及Caspase-3蛋白的表达情况.结果 与对照组比较,经TG作用24 h后,细胞凋亡率增加,并呈浓度依赖性,差异具有统计学意义(P<0.05).TG组在24 h的GRP、CHOP表达均高于对照组,差异具有统计学意义(P<0.05,P<0.01);同时不同刺激组中Caspase-3的表达高于对照组,差异具有统计学意义(P<0.01).结论 毒胡萝卜素可诱导人胚肺成纤维细胞发生凋亡,该凋亡可能与ERS有关.%Aim To study changes of endoplasmic re-tieulum stress relevant protein on the apoptosis in human fetal lung fibroblast cells ( MRC-5 ) and explore the apoptotic effect of thapsigargin ( TG ) on MRC-5 through ER stress activation. Methods Apoptotic rate in MRC-5 cells was observed using flow cytometric a-nalysis. The expression levels of GRP78, CHOP mR-NA was evaluated by RT-PCR. The protein levels of GRP, CHOP, Caspase-3 were assessed by Western blot. Results The apoptotic rates of MRC-5 cells were significantly increased, and were statistically signifi-rncant when compared with the control ( P < 0. 05 ). GRP, CHOP mRNA and proteins levels were significantly increased when the cells were separately treated with TG for 24 h when compared with the control ( P < 0. 05 , P < 0. 01 ). Meanwhile, the protein expression of Caspase-3 in TG was also upregulated. Conclusions The elevated expression of relevant proteins when ERS occurs indicates ERS participates in the process of apoptosis in MRC-5 cells.
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