摘要
目的:探讨KLF11过表达对H2O2诱导的H9c2心肌细胞活力和氧化应激水平的影响及机制。方法:将H9c2细胞分为空白组、H2O2组、空质粒+H2O2组和KLF11+H2O2组。H2O2组使用200μmol/L的H2O2处理细胞6h。空质粒+H2O2组和KLF11+H2O2组细胞分别转染pcDNA3.1及pcDNA3.1-KLF1148h后,使用200μmol/LH2O2处理6h。MTT法检测4组细胞活力,AnnexinV-FITC/PI双染法检测细胞凋亡率,采用SOD及MDA检测试剂盒检测细胞中SOD活力及MDA含量,Westernblot法检测细胞中PI3K、p-AKT和Bcl-2蛋白表达水平。结果:与空白组比较,H2O2组H9c2细胞活力降低,凋亡率增加;细胞中SOD活力降低,MDA含量增加;同时PI3K、p-AKT和Bcl-2蛋白表达下调(P<0.05)。与H2O2组比较,KLF11+H2O2组细胞活力增强,凋亡率降低;细胞中SOD活力增加,MDA含量降低;同时PI3K、p-AKT和Bcl-2蛋白表达上调(P<0.05)。结论:KLF11可能通过激活PI3K/AKT信号通路,抑制氧化应激,提高H2O2诱导的H9c2细胞的活力。
Aim :To investigate the effects of KLF11 overexpression on the viability and oxidative stress of H9c2 cardiomyocytes induced by H 2O 2 and its mechanism. Methods :H9c2 cells were allocated into blank group, H 2O 2 group, pcDNA3.1+H 2O 2 group and pcDNA3.1-KLF11+H2O2 group. The cells in H 2O 2 group were treated with 200 μmol/L H2O2 for 6 hours. The latter two groups were transfected with pcDNA3.1 or pcDNA3.1-KLF11 for 48 hours,respectively, and then treated with H2O2 for 6 hours. Cell viability and apoptotic rate were detected by MTT and Annexin V-FITC/PI staining, SOD activity and MDA content in the cells were detected, and the expressions of PI3K, p-AKT and Bcl-2 proteins were detected by Western blot. Results :Compared with those of the blank group, the viability of cells in H 2O 2 group was inhibited;apoptosis was induced, SOD activity was reduced, MDA content was increased;and the expressions of PI3K, p-AKT and Bcl-2 proteins were down-regulated significantly( P <0.05).While compared with those of the H2O2 group, the viability of cells in pcDNA3.1-KLF11+H2O2 group was induced, apoptosis was inhibited;SOD activity was increased, MDA content was decreased;and the expressions of PI3K, p-AKT and Bcl-2 proteins were up-regulated significantly( P < 0.05 ). Conclusion :KLF11 can improve H9c2 cell viability interfered by H2O2, which may be related to the inhibition of oxidative stress and activation of PI3K/AKT signaling pathway.
作者
王滨
王文华
辛传友
郭龙哲
黄传奇
郜阳
宁文龙
WANG Bin;WANG Wenhua;XIN Chuanyou;GUO Longzhe;HUANG Chuanqi;GAO Yang;NING Wenlong(Emergency Department of Internal Medicine, Qiqihar First Hospital,Qiqihar,Heilongjiang 161000;Department of Rheumatology,Qiqihar First Hospital, Qiqihar,Heilongjiang 161000;Department of Critical Care Medicine, the Second Affiliated Hospital,Ha′erbin Medical University, Ha'erbin, Heilongjiang 150000)
出处
《郑州大学学报（医学版）》
CAS
北大核心
2019年第4期551-555,共5页
Journal of Zhengzhou University(Medical Sciences)
基金
黑龙江省卫生和计划生育委员会科技计划项目(2016-049)
