HBSP抑制缺氧复氧心肌H9C2细胞Omi/HtrA2胞内转位及细胞凋亡  

HBSP inhibits transposition of Omi / HtrA2 and apoptosis of myocardial cells induced by anoxia-reoxygenation

作　　者：游伟[1] 刘映峰[1] 张杰波 林湧滦 缪绯[1] 刘芃[1] 

机构地区：[1]南方医科大学珠江医院心血管内科,广东广州510282

出　　处：《基础医学与临床》2015年第5期615-620,共6页Basic and Clinical Medicine

基　　金：广东省自然科学基金(10151051501000038);广东省科技计划(20130319c)

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/Htr A2胞内转位及细胞凋亡的影响。方法将乳鼠心肌细胞(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/Htr A2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/Htr A2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/Htr A2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/Htr A2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/Htr A2蛋白的胞内转位,抑制caspases通路激活,进而发挥细胞保护作用。Objective To investigate the effect of helix B surface peptide（ HBSP） on transposition of Omi / HtrA 2 and apoptosis of myocardial cells induced by anoxia-reoxygenation. Methods Neonatal rat myocardial cells（ H9C2 cells）were 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 caspase-3 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 / HtrA 2 protein changes in the mitochondrial and cytosolic.Results Compared with the control group,H / R group cell survival rate decreased significantly（ P〈0. 05）,cell supernatant LDH concentration increased significantly,the expression of cleaved caspase-3 increased,the cardiomyocyteapoptosis and the expression of Omi / HtrA 2 protein in the cytoplasm increased significantly（ 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 / HtrA 2 protein in the cytoplasm expression decreased significantly（ P〈0. 05）. Conclusions 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 / HtrA 2 protein in mitochondrial translocation,thus inhibiting the caspases pathway activation.

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 

分 类 号：R542.2[医药卫生—心血管疾病]