摘要
目的构建核呼吸因子-1(NRF-1)过表达的大鼠心肌细胞,观察缺氧条件下细胞增殖和细胞凋亡的变化。方法将大鼠心肌细胞H9C2分为NRF-1过表达组、空载体组和正常对照组,NRF-1过表达组和空载体组分别用NRF-1过表达慢病毒及空载体病毒进行转染,正常对照组不做处理。将三组细胞置于缺氧用三气培养箱中进行缺氧培养(37℃、1%O_2+5%CO_2+94%N2)24 h。采用实时无标记分析(RTCA)技术观察缺氧6、12、24 h时的细胞增殖水平,流式细胞仪检测缺氧24 h时细胞凋亡率,qRT-PCR法检测缺氧24 h时半胱氨酸天冬氨酸蛋白酶3(caspase-3)mRNA的相对表达量。结果缺氧12、24 h时,NRF-1过表达组细胞增殖水平高于正常对照组和空载体组;缺氧24 h时,与空载体组和正常对照组比较,NRF-1过表达组细胞凋亡水平降低,且caspase-3 mRNA表达水平较低(P均<0.05)。结论 NRF-1过表达可以降低缺氧条件下心肌细胞的凋亡,从而提高心肌细胞在缺氧条件下的存活能力。
Objective To observe the changes of cell proliferation and apoptosis of rat cardiomyocytes with nuclear respiratory factor-1（ NRF-1） over-expression under hypoxia. Methods The rat cardiomyocytes H9C2 were divided into the NRF-1 over-expression group,empty vector group,and control group. The cells in the NRF-1 over-expression group and empty vector group were transfected with NRF-1 overexpressed lentivirus and empty vector lentivirus,respectively,and the control group was not treated. Cells of the three groups were placed into three gas incubator for anoxic culture（ 37 ℃,1%O_2＋ 5% CO_2＋ 94% N2） for 24 h. The real-time cell assay（ RTCA） technology was used to observe cell proliferation level under normal and hypoxic conditions. Flow cytometry was used to detect the apoptosis rate,and qRT-PCR was applied to detect the relative expression of caspase-3 mRNA. Results At 24 and 48 h after hypoxia,the cells of NRF-1 over-expression group had a higher proliferation level than the control group and empty vector group. At 24 h after hypoxia,the apoptosis level of NRF-1 over-expression group decreased significantly,and the expression level of caspase-3 mRNA was lower as compared with that of the empty vector group and the control group（ both P〈0. 05）. Conclusion Over-expression of NRF-1 may reduce the apoptosis of cardiomyocytes under hypoxia condition,and enhance the viability of cardiomyocytes.
作者
孙红丽
杨继辉
牛楠
朱明星
赵巍
宋熔
SUN Hongli;YANG Jihui;NIU Nan;ZHU Mingxing;ZHAO Wei;SONG Rong(Ningxia Medical University,Yinchuan 750004,Chin)
出处
《山东医药》
CAS
2018年第26期38-40,共3页
Shandong Medical Journal
