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.
出处
《基础医学与临床》
CSCD
北大核心
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)
