缺氧对大鼠心肌细胞损伤的影响及机制

发布时间：2018-03-19 08:13

本文选题：缺氧　切入点：线粒体损伤　出处：《中国老年学杂志》2017年05期 　论文类型：期刊论文

【摘要】：目的探讨缺氧对大鼠心肌细胞系H9C2的损伤及机制。方法在1%O_2+5%CO_2+94%N_2条件下培养大鼠心肌细胞株H9C2 3、6、12、24、48 h,并以正常大鼠心肌细胞株作对照,使用台盼蓝进行细胞计数;CCK-8法检测细胞存活率;实时监测心肌细胞生长状态;透射电镜观察细胞内部结构;活性氧检测试剂盒检测胞内活性氧(ROS);Western印迹法检测线粒体色素C蛋白表达水平;流式细胞术检测细胞凋亡率;乳酸脱氢酶(LDH)试剂盒检测细胞坏死程度。结果缺氧状态下,随着缺氧时间的不断增加,大鼠心肌细胞株H9C2的细胞生长、存活率均明显降低、细胞形态明显改变,细胞器结构明显损伤,ROS水平升高,线粒体细胞色素释放增加,细胞凋亡率明显增加,LDH的释放率明显增高。结论缺氧会造成大鼠心肌细胞的生长速度减慢、形态改变及心肌线粒体损伤,最终导致细胞凋亡和细胞坏死。
[Abstract]:Objective to investigate the damage and mechanism of hypoxia on rat myocardial cell line H9C2. Methods cultured rat cardiomyocyte line H9C23, H99C2 for 2448 hours, and normal rat cardiomyocytes as control. Trypan blue was used to count cells and CCK-8 method was used to detect cell survival rate, real-time monitoring of cardiac myocyte growth status, transmission electron microscope observation of cell internal structure. The expression of mitochondrial pigment C protein was detected by Western blotting, apoptosis rate was detected by flow cytometry, necrosis degree was detected by LDH kit. With the increase of hypoxia time, the cell growth and survival rate of rat cardiomyocyte line H9C2 decreased significantly, the cell morphology changed obviously, the level of Ros and the release of cytochrome from mitochondria increased significantly. Conclusion hypoxia can decrease the growth rate of rat cardiomyocytes, change the morphology and damage myocardial mitochondria, and eventually lead to apoptosis and cell necrosis.
【作者单位】：宁夏医科大学基础医学院;宁夏医科大学科技中心;宁夏医科大学临床医学院;中国人民解放军第五医院重症医学科;
【基金】：国家自然科学基金项目资助(81360042)
【分类号】：R54

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