DDAH2抑制低氧诱导大鼠心肌细胞H9c2(2-1)凋亡

基础医学与临床 ›› 2013, Vol. 33 ›› Issue (1): 28-32.

DDAH2抑制低氧诱导大鼠心肌细胞H9c2(2-1)凋亡

吴斌¹,李旭光²,张玫¹,罗通行¹,黄亨健¹,王兰兰¹

1. 四川大学华西医院
2. 上海交通大学附属上海第一人民医院

收稿日期:2012-02-02 修回日期:2012-05-03 出版日期:2013-01-05 发布日期:2012-12-25
通讯作者: 王兰兰 E-mail:wchcbc@yahoo.com.cn

DDAH2 overexpression inhibits hypoxia-induced apoptosis of rat cardiomyocytes

Received:2012-02-02 Revised:2012-05-03 Online:2013-01-05 Published:2012-12-25
Contact: Lan-lan WANG E-mail:wchcbc@yahoo.com.cn

摘要/Abstract

摘要： 目的探讨过表达二甲基精氨酸二甲胺水解酶-Ⅱ（DDAH2）对低氧诱导的大鼠心肌细胞凋亡的抑制作用和是否通过内皮型一氧化氮合酶（eNOS）活化来保护心肌细胞。方法低氧诱导大鼠心肌细胞H9c2(2-1)36h从而诱导凋亡，同时转染DDAH2真核细胞表达质粒，用WST-1细胞毒试验和流式细胞术检测细胞凋亡,Western blot法检测eNOS磷酸化。结果低氧组心肌细胞存活率显著性降低；过表达DDAH2基因使低氧下心肌细胞存活率明显地上升。DDAH2过表达组心肌细胞凋亡率为17.38%±1.52%，显著性低于低氧组（27.34%±1.33%）（P<0.05）。转染DDAH2基因可以显著上调低氧组心肌细胞的eNOS磷酸化水平；过表达DDAH2基因的低氧组用eNOS抑制剂L-NAME干预后,凋亡水平明显上升。结论过表达DDAH2基因上调eNOS磷酸化抑制低氧所致的大鼠心肌细胞H9c2（2-1）凋亡。

关键词: 二甲基精氨酸二甲胺水解酶-Ⅱ,缺氧,心肌细胞,细胞凋亡

Abstract: Objective To investigate the inhibitory effect of DDAH2 on the apoptosis induced by hypoxia in rat cardiomyocytes through phosphorylation of eNOS. Methods Cell apoptosis was induce by hypoxia within 36hour and was determined by wst-1 and flow cytometry. The phosphorylation of eNOS was examined by western-blot. Results Survival rate of hypoxia group was significantly decreased compared with control group; overexpression DDAH2 induces survival of myocardial cells markedly. After transfection of DDAH2 gene to cardiomyocytes, hypoxia-induced apoptosis significantly declined (17.38%±1.52% vs 27.34%±1.33%, P<0.05). DDAH2 gene transfection significantly increased level of phosphorylation of eNOS in hypoxia of myocardial cells. eNOS inhibitor, L-NAME alleviated the anti-apoptosis effect of overexpression-DDAH2 significantly in H9c2(2-1).Conclusion Overexpression DDAH2 inhibits the apoptosis effects of hypoxia on rat cardiomyocytes H9c2 (2-1) by upregulation of phosphorylation of eNOS.

Key words: DDAH-2, hypoxia, cardiomyocytes, apoptosis

中图分类号:

R318.11

吴斌李旭光张玫罗通行黄亨健王兰兰. DDAH2抑制低氧诱导大鼠心肌细胞H9c2(2-1)凋亡[J]. 基础医学与临床, 2013, 33(1): 28-32.