摘要
目的:评价细胞穿透肽4即蛋白转导结构域4(PTD4)介导的铜锌超氧化物歧化酶(Cu/Zn SOD)对大鼠心肌细胞缺氧复氧损伤(HRI)的影响。方法:用厌氧(85%N2、10%H2、5%CO2)培养箱制作大鼠心肌细胞(H9C2)HRI模型,设置HRI组(HRI的细胞培养液中不加任何处理因素)、HRI+Cu/Zn SOD组(加入10μmol·L-1Cu/Zn SOD)、HRI+PTD4-Cu/Zn SOD组(10μmol·L-1PTD4-Cu/Zn SOD),另外以正常培养心肌细胞作为正常对照组,孵育30 min后,末端脱氧核苷酸转移酶(Td T)介导的d UTP缺口末端标记(TUNEL)技术检测心肌细胞凋亡,蛋白印迹法检测心肌细胞Bcl-2、Bax蛋白的表达。结果:HRI+PTD4-Cu/Zn SOD组心肌细胞凋亡指数[(10.20±2.77)%]比HRI组[(28.40±2.41)%]明显降低,与HRI组比较,HRI+PTD4-Cu/Zn SOD融合蛋白能显著增加Bcl-2蛋白表达,降低Bax蛋白表达。结论:PTD4-Cu/Zn SOD融合蛋白可以减轻大鼠心肌细胞的HRI。
Objective： To investigate whether the cell- penetrating peptide, protein transduction domain 4 （PTD4） mediates the protective effects of copper-zinc superoxide dismutase （Cu/Zn SOD） on hypoxiafreoxygenation injury （HRI） in rat myocardial cells. Methods： Myocardial cell H9C2 cultured in vitro were randomized into 4 groups： normal group, HRI group, HRI-treated H9C2 cells with Cu/Zn SOD （HRI ＋ Cu/Zn SOD group）, HRI-treated H9C2 cells with PTD4 and Cu/Zn SOD （ HRI ＋ PTD4- Cu/Zn SOD group ）. HRI was achieved by exposing eardiomyocytes to 4 h hypoxia followed by 2 h reoxygenation. Cardiomyocyte apoptotic index, Bax and Bcl- 2 in cardiomyocytes were detected with TUNEL and Western blot. Results： HRI ＋ PTD4-Cu/Zn SOD reduced apoptosis of cardiomyocytes induced by hypoxia/reoxygenation injury. Compared with HRI group, HRI ＋ PTD4- Cu/Zn SOD group significantly reduced the expression of Bax and increase the expression of Bcl-2. Conclusion： Cu/Zn SOD mediated by cell penetrating peptide PTIM can attenuate hypoxia/reoxygenation injury in rat myocardial cells.
出处
《现代医学》
2017年第10期1396-1399,共4页
Modern Medical Journal
基金
国家自然科学基金资助项目(81171783)
