基础医学与临床 ›› 2015, Vol. 35 ›› Issue (5): 615-620.

• 研究论文 • 上一篇    下一篇

HBSP抑制缺氧复氧心肌H9C2细胞Omi/HtrA2胞内转位及细胞凋亡

游伟1,刘映峰1,张杰波2,林湧滦2,缪绯1,刘芃1   

  1. 1. 南方医科大学珠江医院心血管内科
    2. 南方医科大学
  • 收稿日期:2014-11-04 修回日期:2015-01-22 出版日期:2015-05-05 发布日期:2015-04-28
  • 通讯作者: 刘芃 E-mail:sxlp9069@gmail.com
  • 基金资助:
    广东省自然科学基金;广东省科技计划项目

Effect of HBSP on transposition of Omi/HtrA2 and apoptosis of myocardial cells Induced by Anoxia- reoxygenation

  • Received:2014-11-04 Revised:2015-01-22 Online:2015-05-05 Published:2015-04-28

摘要: 目的 探讨促红细胞生成素衍生肽(HBSP)对缺氧复氧心肌细胞Omi/HtrA2胞内转位及细胞凋亡的影响。方法 将乳鼠心肌细胞株(H9C2 细胞)分为对照(Ctrl)组、缺氧复氧(H/R)组、HBSP组和EPO组。培养结束后MTS法检测细胞存活率 ,酶标仪检测细胞上清液中LDH释放率,Western Blot检测细胞内cleaved caspase-3表达,TUNEL法检测心肌细胞凋亡;分离H9C2细胞胞质及线粒体,Western blot分别检测线粒体及细胞质Omi/HtrA2表达。结果 与Ctrl组相比,H/R 组细胞存活率下降(P<0.05),LDH释放、cleaved caspase-3表达、心肌细胞凋亡及Omi/HtrA2胞内转位均明显上升(P<0.05);与H/R组相比,EPO组的细胞存活率升高(P<0.05),LDH释放降低、cleaved caspase-3表达减弱、心肌细胞凋亡减少、Omi/HtrA2线粒体转位明显减少(P<0.05);随着HBSP浓度的增加,各组细胞存活率逐渐上升,LDH释放、cleaved caspase-3表达、心肌细胞凋亡及Omi/HtrA2胞内转位率逐渐下降 (P<0.05)。结论 HBSP具有与EPO类似的保护作用,可抑制经H / R诱导的心肌细胞凋亡,其机制可能是通过减少Omi/HtrA2蛋白的胞内转位,抑制Caspases通路激活,进而发挥细胞保护作用。

关键词: 缺氧复氧, 促红细胞生成素衍生肽, 促红细胞生成素, 细胞凋亡, Omi/HtrA2

Abstract: Objective To investigate the effect of helix B surfacepeptide(HBSP) on transposition of Omi/HtrA2 and apoptosis of myocardial cells Induced by Hypoxia- reoxygenation. Methods neonatal rat myocardial cells (H9C2 cells) was used to be research object, the cells were divideded into 4 groups:control group, H/R group, HBSP group and EPO group. The cell viability was measured by MTS assay, the concentration of LDH of cell supernatant was assessed by ELISA, the intracellular expression changes of cleaved Caspase3 was detected by Western blot. apoptosis of cardiomyocytes was detected by TUNEL.Using Mitochondria/cytosol Isolation Kit to isolate Mitochondria from cytosol, Western blot was used to detect the expression of Omi/HtrA2 protein changes in the mitochondrial and cytosolic.Results Compared with the control group, H/R group cell survival rate decreased significantly, cell supernatant LDH concentration increased significantly,the expression of cleaved caspase-3 was increased, the cardiomyocyte apoptosis and the expression of Omi/HtrA2 protein in the cytoplasm is increased,the differences were statistically significant (P<0.05); compared with H/R group, EPO and HBSP group cell survival rate increased, cell supernatant concentrations of LDH、cleaved caspase-3、cardiomyocyte apoptosis and the Omi/HtrA2 protein in the cytoplasm expression decreased, the difference was statistically significance (P<0.05).conclusion H / R can induce apoptosis of myocardial cell,which can be reduced by EPO or HBSP,its mechanism may be related to the inhibition of Omi/HtrA2 protein in mitochondrial translocation, thus inhibiting the Caspases pathway activation.

Key words: hypoxia-reoxygenation , HBSP , EPO , apoptosis , Omi/HtrA2