miR-499a-5p通过靶向APC减轻氧化应激对心肌细胞的损伤  

miR-499a-5p attenuates oxidative stress damage to cardiomyocytes by targeting APC

作　　者：武国利[1] 马竞 WU Guoli;MA Jing(Baoding First Central Hospital,Baodingy Hebei 071000,China;Affiliated Hospital of Hebei Universityy Baoding,Hebei 071000,China)

机构地区：[1]保定市第一中心医院,河北省保定市071000 [2]河北大学附属医院,河北省保定市071000

出　　处：《中国动脉硬化杂志》2020年第6期501-506,共6页Chinese Journal of Arteriosclerosis

基　　金：保定市科技计划项目(1951ZF058)。

摘　　要：目的研究miR-499a-5p对过氧化氢(H2O2)诱导的心肌细胞H9c2增殖、凋亡的影响,并探讨其作用机制。方法用细胞计数试剂盒(CCK-8)检测不同浓度H2O2(100、200、400、800μmol/L)处理6 h的H9c2细胞的存活率,筛选400μmol/L H2O2处理的H9c2细胞作为模型组。将模型组细胞分为miR-NC组、miR-499a-5p组、si-NC组、si-APC组、miR-499a-5p+pcDNA组、miR-499a-5p+pcDNA-APC组,用流式细胞术、免疫印迹(Western blot)、酶联免疫吸附试验(ELISA)检测各组细胞的存活率、凋亡率、活性氧(ROS)、超氧化物歧化酶(SOD)、丙二醛(MDA)及增殖凋亡相关蛋白增殖细胞核抗原(PCNA)、细胞周期蛋白依靠性激酶抑制剂(P21)、B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关的X基因(Bax)的表达。结果 H2O2(100、200、400、800μmol/L)呈浓度依赖性抑制H9c2细胞的存活,最适浓度为400μmol/L。模型组细胞中miR-499a-5p表达显著降低,APC表达显著升高;过表达miR-499a-5p、抑制APC均可明显减轻H2O2诱导的H9c2细胞的增殖抑制、凋亡促进和氧化应激作用,并且miR-499a-5p还可靶向抑制APC。过表达APC逆转了miR-499a-5p对H2O2诱导的心肌细胞损伤。结论 miR-499a-5p可调控H2O2诱导的心肌细胞增殖、凋亡和氧化应激,其机制与靶向抑制APC有关,将可为氧化应激引起的心肌细胞损伤的治疗提供新靶点。Aim To investigate the effect of mi R-499 a-5 p on hydrogen peroxide( H2 O2)-induced proliferation and apoptosis of cardiomyocytes H9 c2,and to explore its mechanism. Methods Cell viability kit( CCK-8) was used to detect the survival rate of H9 c2 cells treated with different concentrations of H2 O2( 100,200,400,800 μmol/L) for 6 h,and 400 μmol/L treated H9 c2 cells were selected as a model group. The model group cells were divided into mi R-NC group,mi R-499 a-5 p group,si-NC group,si-APC group,mi R-499 a-5 p+pc DNA group,mi R-499 a-5 p+pc DNA-APC group.Flow cytometry,western blotting( Western blot) and enzyme linked immunosorbent assay( ELISA) were used to detect the survival rate,apoptosis rate,reactive oxygen species( ROS),superoxide dismutase( SOD),melondialodehyde( MDA)and proliferation-related protein expression proliferating cell nuclear antigen( PCNA),cyclins depend on kinase inhibitors( P21),B lymphoblastoma-2( Bcl-2),X gene associated with Bcl-2( Bax) of each group. Results H2 O2( 100,200,400,800 μmol/L) inhibited the survival of H9 c2 cells in a concentration-dependent manner with an optimal concentration of 400 μmol/L. The expression of mi R-499 a-5 p was significantly decreased and the expression of APC was significantly increased in the model group. Overexpression of mi R-499 a-5 p and inhibition of APC significantly attenuated H2O2-induced proliferation inhibition,apoptosis promotion and oxidation stress of H9 c2 cells. mi R-499 a-5 p can also target inhibition of APC. Overexpression of APC reversed the damage of H2O2-induced cardiomyocytes by mi R-499 a-5 p.Conclusions mi R-499 a-5 p can regulate the proliferation,apoptosis and oxidative stress of cardiomyocytes induced byH2O2. The mechanism is related to the targeted inhibition of APC,which will provide a new target for the treatment of myocardial cell injury induced by oxidative stress.

关 键 词：miR-499a-5p APC 心肌细胞损伤 

分 类 号：R542.2[医药卫生—心血管疾病]