色素上皮衍生因子对缺氧条件下H9C2心肌细胞保护作用

发布时间：2018-06-01 20:04

本文选题：细胞凋亡 + 缺氧　；参考：《重庆医学》2017年26期

【摘要】：目的探讨色素上皮衍生因子(PEDF)对H9C2心肌细胞在低氧无血清条件下的保护作用及其可能的机制。方法体外培养H9C2细胞进行低氧无血清处理,将细胞分为对照组(H9C2)、缺氧组(缺氧+H9C2)、PEDF组(缺氧+H9C2+PEDF)、残粒体分裂抑制剂(Medivi-1)组(缺氧+H9C2+Mdivi-1)。TUNEL染色检测H9C2心肌细胞凋亡率;Western blot检测动力相关蛋白1(Drp1)、活化半胱天冬酶-3(Cleaved-Caspase3)的蛋白水平;电镜及MitoTracker Red检测线粒体形态;采用线粒体膜电位检测试剂盒(JC-1)检测线粒体膜电位,MitoSOXTM检测线粒体活性氧簇(ROS)水平。结果缺氧诱导H9C2细胞线粒体分裂,缺氧组(6h)与对照组比较差异有统计学意义(P0.05),PEDF减少缺氧条件下线粒体分裂,PEDF组与缺氧组(6h)比较差异有统计学意义(P0.05),PEDF和Mdivi-1可以减少缺氧条件下(24h)细胞凋亡,与缺氧组(24h)比较差异有统计学意义(P0.05)。结论 PEDF通过抑制缺氧条件下H9C2细胞线粒体分裂减少细胞凋亡。
[Abstract]:Objective to investigate the protective effect of pigment epithelium-derived factor (PEDF) on H9C2 cardiomyocytes under hypoxic and serum-free conditions and its possible mechanism. Methods H9C2 cells cultured in vitro were treated with hypoxia and serum free. The cells were divided into two groups: control group (H9C2P), hypoxia group (hypoxia H9C2P2PEDF group) group (hypoxia H9C2 PEDF group, residual granulocyte mitosis inhibitor medivi-1 group) group (H9C2 cardiomyocyte apoptosis rate was detected by anoxic H9C2 Mdivi-1).TUNEL staining and H9C2 cardiomyocyte apoptosis rate was detected by Western blot to detect the protein level of dynamic-associated protein 1, activated cysteone asparagase -3Cvelead-Caspase3). The mitochondrial morphology was detected by electron microscope and MitoTracker Red, and the mitochondrial membrane potential (MitoSOXTM) was detected by using mitochondrial membrane potential detection kit (JC-1), and the mitochondrial reactive oxygen species (Ros) level was detected by mitochondrial membrane potential detection kit (JC-1). Results hypoxia induced mitochondrial division in H9C2 cells. There was significant difference between the hypoxia group and the control group at 6 h) the difference was statistically significant (P 0.05) PEDF could reduce the apoptosis of mitochondria mitosis in the hypoxia group and the hypoxia group at 6 h) the P0.05 Mdivi-1 and PEDF could reduce the apoptosis of the cells under hypoxia for 24 h. Compared with hypoxia group for 24 h, the difference was statistically significant (P 0.05). Conclusion PEDF reduces apoptosis by inhibiting mitochondrial mitosis in H9C2 cells under hypoxia.
【作者单位】：徐州医科大学附属医院心胸外科;徐州医科大学生物化学与分子生物学研究中心;徐州医科大学形态学科研实验中心;
【基金】：国家自然科学基金资助项目(81270173) 江苏省卫生厅科技计划项目青年基金(Q201407)
【分类号】：R54

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