毒胡萝卜素对人胚肺成纤维细胞凋亡影响及部分机制的初步探讨  

Effect and mechanism of thapsigargin in apoptosis in MRC-5 cells

作　　者：王亚丽[1] 李俊[1] 黄成[1] 黄艳[1] 于明哲[1] 任丹阳[1] 徐涛[1] 

机构地区：[1]安徽医科大学药学院,安徽合肥230032

出　　处：《中国药理学通报》2012年第9期1281-1285,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81102493;30973917);安徽省自然科学基金资助项目(No 090413134)

摘　　要：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/enhancerbinding 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、CHOPmRNA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 retieulum 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 analysis. The expression levels of GRP78, CHOP mRNA 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 signifiant 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.

关 键 词：内质网应激 人胚肺成纤维细胞 毒胡萝卜素 GRP CHOP Caspase-3 

分 类 号：R322.35[医药卫生—人体解剖和组织胚胎学] R329.24[医药卫生—基础医学]