基础医学与临床 ›› 2010, Vol. 30 ›› Issue (9): 956-960.

• 研究论文 • 上一篇    下一篇

过氧亚硝酸盐增强人脑血管平滑肌细胞PDCD4基因表达

李林 陈晓锐 宋莎莎 焦玉莲 马春燕 鞠远荣 李建峰   

  1. 山东大学附属省立医院 山东大学附属省立医院
  • 收稿日期:2009-11-09 修回日期:2010-01-24 出版日期:2010-09-05 发布日期:2010-09-05
  • 通讯作者: 李建峰

Peroxynitrite Increases the Expression of PDCD4 Gene in Human Cerebral Vascular Smooth Muscle Cells

Lin LI, Xiao-rui CHEN, Sha-sha SONG, Yu-lian JIAO, Chun-yan MA, Yuan-rong JU, Jian-feng LI   

  1. Provincial Hospital affiliated to Shandong University Provincial Hospital affiliated to Shandong University
  • Received:2009-11-09 Revised:2010-01-24 Online:2010-09-05 Published:2010-09-05
  • Contact: Jian-feng LI

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PDCD4 mRNA及蛋白表达显著增加。结论 PDCD4基因可能在ONOO-诱导HCVSMCs凋亡的信号转导通路中起到关键作用。

关键词: ONOO-, PDCD4, 中枢血管平滑肌细胞, 细胞凋亡

Abstract: Objective This study was designed to investigate the change of PDCD4 gene expression in cerebral vascular smooth muscle cells (HCVSMCs) in response to peroxynitrite (ONOO-). Methods HCVSMCs were cultured in vitro and subjected to different concentrations of ONOO-. Cell viability was measured by MTT assay. The morphological changes were assessed 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 revealed via the apoptotic pathway. Under the stress by ONOO-, the expression of PDCD4 gene was apparently increased at both mRNA and protein levels. Conclusions PDCD4 gene may play a pivotal role underlying the ONOO--induced apoptosis in HCVSMCs.

Key words: Peroxynitrite, PDCD4 gene, cerebral vascular smooth cells, Apoptosis

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