过氧亚硝酸盐增强人脑血管平滑肌细胞PDCD4基因表达  

Peroxynitrite increases the expression of PDCD4 gene in human cerebral vascular smooth muscle cells

作　　者：李林[1] 陈晓锐[2] 宋莎莎[1] 焦玉莲[1] 马春燕[1] 鞠远荣[3] 李建峰[1] 

机构地区：[1]山东大学附属省立医院中心实验室,山东济南250021 [2]滨州医学院医学人文学院,山东烟台264003 [3]山东大学附属省立医院ICU室,山东济南250021

出　　处：《基础医学与临床》2010年第9期956-960,共5页Basic and Clinical Medicine

基　　金：山东省科技厅基金(2006GG2202)

摘　　要：目的探讨过氧亚硝酸盐(ONOO-)诱导人脑血管平滑肌细胞(HCVSMCs)凋亡过程中PDCD4基因表达的变化。方法以体外培养的HCVSMCs系为实验材料,观察不同浓度的ONOO-对HCVSMCs的作用。采用噻唑蓝(MTT)比色法测定细胞生存率,流式细胞仪测定细胞凋亡率,吖啶橙染色、Ho33342/PI荧光双染进行形态学观察,RT-PCR及Western blot检测PDCD4的mRNA及蛋白表达。结果ONOO-作用HCVSMCs 24 h,显著抑制细胞增殖,并诱导细胞凋亡,同时引起PDCD4mRNA及蛋白表达显著增加。结论PDCD4基因可能在ONOO-诱导HCVSMCs凋亡的信号传导通路中起到关键作用。Objective To investigate the change of PDCD4 gene expression in cerebral vascular smooth muscle cells（HCVSMCs） as a response to peroxynitrite（ONOO-）.Methods HCVSMCs were cultured in vitro and exposed to different concentrations of ONOO-.Cell viability was measured by MTT assay.The morphological changes were by acridine orange staining as well as Ho33342/PI double staining to determine whether cell experienced apoptosis.The apoptotic rate was determined by flow cytometry.In particular,the expression of PDCD4 gene was examined by the means of RT-PCR and Western blot simultaneously.Results Direct exposure of HCVSMCs to ONOO-was able to block the cell proliferation,which was further demonstrated via the apoptotic pathway.Under the stress by ONOO-,the expression of PDCD4 gene was apparently increased at both mRNA and protein levels.Conclusion PDCD4 gene may play a pivotal role underlying the ONOO——induced apoptosis in HCVSMCs.

关 键 词：过氧亚硝酸盐 PDCD4 人脑血管平滑肌细胞 细胞凋亡 

分 类 号：R743.3[医药卫生—神经病学与精神病学]