[1]吴庆,姚浩旗,程阔菊,等.Carabin过表达对人心室肌细胞AC16氧糖剥夺后凋亡的影响[J].第三军医大学学报,2018,40(22):2068-2073.
 WU Qing,YAO Haoqi,CHENG Kuoju,et al.Effect of Carabin overexpression on oxygenglucose deprivation-induced apoptosis in cultured human ventricular myocytes[J].J Third Mil Med Univ,2018,40(22):2068-2073.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第22期
页码:
2068-2073
栏目:
基础医学
出版日期:
2018-11-30

文章信息/Info

Title:
Effect of Carabin overexpression on oxygenglucose deprivation-induced apoptosis in cultured human ventricular myocytes
作者:
吴庆姚浩旗程阔菊唐建芳黄河陈建李洪杨天德
陆军军医大学(第三军医大学)第二附属医院麻醉科;陆军军医大学(第三军医大学)高原军事医学系
Author(s):
WU Qing YAO Haoqi CHENG Kuoju TANG Jianfang HUANG He CHEN Jian LI Hong YANG Tiande

Department of Anesthesiology, Second Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400037;  Faculty of High Altitude Military Medicine, Army Medical University (Third Military Medical University) , Chongqing, 400038, China

关键词:
人心室肌AC16细胞Carabin氧糖剥夺细胞凋亡
Keywords:
human ventricular myocytes cells AC16 Carabin oxygenglucose deprivation cell apoptosis
分类号:
R322.11; R394-33; R394.2
文献标志码:
A
摘要:

目的探讨Carabin基因过表达对氧糖剥夺(oxygen glucose deprivation, OGD)诱导的人心室肌细胞株AC16凋亡的作用。方法构建Carabin过表达及空载稳定细胞株,将AC16细胞分为空载常氧组、Carabin过表达常氧组、空载OGD 24 h组和Carabin过表达OGD 24 h组。空载常氧组及Carabin过表达常氧组置于正常条件下培养;空载OGD 24 h组和Carabin过表达OGD 24 h组置于Hanks液中放入缺氧恒温细胞培养箱培养24 h(1%O2-5%CO2-94%N2)。采用倒置显微镜观察细胞形态变化;流式细胞术检测心肌细胞凋亡率及存活率;TUNEL染色检测细胞凋亡情况;通过酶标仪检测心肌细胞凋亡蛋白酶Caspase-3和心肌细胞损伤标志物LDH的活性;Western blot检测细胞凋亡蛋白Caspase-3的表达水平。结果AC16细胞OGD 24 h后,细胞内Carabin的mRNA及蛋白水平显著降低。与空载常氧组比较,空载OGD 24 h组及Carabin过表达OGD 24 h组的细胞凋亡率、Caspase3活性、LDH活性及Caspase-3蛋白表达水平均显著升高(P<0.05)。与空载OGD 24 h组比较,Carabin过表达OGD 24 h组的细胞凋亡率、Caspase-3活性、LDH活性及Caspase-3蛋白表达水平显著降低(P<0.05)。结论Carabin过表达能显著减轻OGD诱导后AC16细胞的凋亡。

Abstract:

ObjectiveTo investigate the effect of Carabin overexpression on apoptosis of human ventricular myocytes induced by oxygenglucose deprivation (OGD) in vitro. MethodsHuman ventricular myocyte AC16 cell line was transfected with a lentiviral vector carrying Carabin gene to induce Carabin overexpression or with a negative control vector. The transfected cells were cultured for 24 h under routine conditions or under OGD conditions (in Hank’s solution in a hypoxia incubator with 1% O2, 5% CO2, and 94% N2). The changes in the morphology of AC16 cells were observed under an inverted microscope, and flow cytometry was used to determine the apoptosis rate and survival rate of the cells. The apoptotic cells were detected with TUNEL staining, and the activity of Caspase-3 and LDH in the cells were evaluated with enzyme labeling. Western blotting was performed to determine the protein expression of Caspase-3 in the cells in different culture conditions. ResultsOGD for 24 h significantly lowered the mRNA and protein levels of Carabin in AC16 cells. In the cells transfected with Carabin-overexpressing vector and the negative control vector, OGD for 24 h resulted in significantly increased cell apoptosis rate, Caspase-3 activity, LDH activity, and Caspase-3 protein expression as compared with the routinely cultured cells transfected with the negative control vector (P<0.05). Compared with the negative control cells, the cells overexpressing Carabin were more resistant to OGD and showed significantly lower apoptotic rate, Caspase-3 and LDH activities, and Caspase-3 protein expression after OGD for 24 h (P<0.05). ConclusionThe overexpression of Carabin can significantly reduce the apoptosis of AC16 cells induced by OGD.

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更新日期/Last Update: 2018-12-03