甘油醛-3-磷酸脱氢酶核转位与高糖应激诱导的H9c2细胞凋亡密切相关

中国临床药理学与治疗学 ›› 2011, Vol. 16 ›› Issue (4): 374-379.

甘油醛-3-磷酸脱氢酶核转位与高糖应激诱导的H9c2细胞凋亡密切相关

孙传波, 高景红, 胡亮, 张竞文, 魏立, 李庆平

南京医科大学基础医学院药理学系,江苏省心血管病分子干预重点实验室,南京 210029,江苏

收稿日期:2011-04-04 修回日期:2011-04-21 发布日期:2011-06-22
通讯作者: 李庆平,女,教授,博导,研究方向:心血管药理学。Tel: 025-86862883 E-mail: qingpingli@yahoo.com.cn
作者简介:孙传波,女,硕士研究生,研究方向:心血管药理学。

High glucose-induced apoptosis of H9c2 cells is closely related to GAPDH nuclear translocation

SUN Chuan-bo,GAO Jing-hong, HU Liang, ZHANG Jing-wen, WEI Li, LI Qing-ping

Department of Pharmacology, Nanjing Medical University, Key Laboratory of Cardiovascular Diseace and Molecular Intervention, Nanjing 210029, Jiangsu, China

Received:2011-04-04 Revised:2011-04-21 Published:2011-06-22

摘要/Abstract

摘要： 目的: 探讨高糖应激是否引起心肌细胞中甘油醛-3-磷酸脱氢酶(GAPDH)核转位以及GAPDH核转位是否与细胞凋亡有关。 方法: 高糖培养H9c2细胞 48 h 后,收集细胞进行细胞凋亡的测定,用DCFH-DA荧光探针测定细胞中活性氧簇(ROS)的水平,用Western Blotting方法测定细胞质和细胞核中GAPDH以及细胞中诱导型一氧化氮合酶(iNOS)的表达,用细胞免疫荧光的方法测定GAPDH的核转位情况。 结果: 与低糖培养组相比,高糖培养组H9c2细胞凋亡率增高,ROS生成和iNOS表达增多,GAPDH由细胞质向细胞核转位;而线粒体呼吸链复合体I抑制剂鱼藤酮和iNOS抑制剂1400W均阻断GAPDH的转位,并减少高糖诱导的H9c2细胞的凋亡。 结论: 高糖诱导H9c2细胞中GAPDH由细胞质向细胞核转位;鱼藤酮和1400W可抑制高糖诱导的细胞凋亡,这一作用与抑制GAPDH核转位有关。

关键词: H9c2细胞, 高糖, 细胞凋亡, 甘油醛-3-磷酸脱氢酶核转位, 一氧化氮合酶

Abstract: AIM: To investigate whether high glucose induces GAPDH nuclear translocation and whether the GAPDH nuclear translocation is related to high glucose-induced apoptosis of H9c2 cells. METHODS: H9c2 cells were cultured for 48 h in high glucose condition, and then cells were collected for apoptosis determination. The generation of reactive oxygen species (ROS) was determined. The expression of GAPDH and iNOS was measured by western blotting. The translocation of GAPDH was determined by immunofluorescence. RESULTS: High glucose induced apoptosis of H9c2 cells. High glucose induced oxidative stress in H9c2 cells and translocation of GAPDH to nucleus. Rotenone and iNOS inhibitor 1400W protected H9c2 cells from death by inhibiting GAPDH translocation to nucleus. CONCLUSION: High glucose induces GAPDH translocation from cytoplasm to nucleus. The inhibition of high glucose-induced apoptosis of H9c2 cells is closely related to GAPDH nuclear translocation.

Key words: H9c2 cells, High glucose, Cell apoptosis, GAPDH translocation, Nitric oxide synthase

中图分类号:

R965.2

孙传波, 高景红, 胡亮, 张竞文, 魏立, 李庆平. 甘油醛-3-磷酸脱氢酶核转位与高糖应激诱导的H9c2细胞凋亡密切相关[J]. 中国临床药理学与治疗学, 2011, 16(4): 374-379.

SUN Chuan-bo,GAO Jing-hong, HU Liang, ZHANG Jing-wen, WEI Li, LI Qing-ping. High glucose-induced apoptosis of H9c2 cells is closely related to GAPDH nuclear translocation[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2011, 16(4): 374-379.

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