miR-6216对缺氧/复氧H9C2心肌细胞损伤的影响  

The effect of lentivirus mediated mir-6216 on the injury of hypoxia/reoxygenation cardiomyocytes

作　　者：冯彦景[1] 李艳君[1] 殷智晔[1] 贾涛 FENG Yanjing;LI Yanjun;YIN Zhiye;JIA Tao(Hebei Chest Hospital,Shijiazhuang 050048,China;Shijiazhuang Luancheng People's Hospital,Shijiazhuang 051430,China)

机构地区：[1]河北省胸科医院,河北石家庄050048 [2]石家庄市栾城人民医院,河北石家庄051430

出　　处：《西部医学》2020年第10期1454-1460,共7页Medical Journal of West China

摘　　要：目的探讨miR-6216对缺氧/复氧(H/R)诱导的心肌细胞损伤的影响。方法体外培养大鼠胚胎心肌细胞H9C2,构建H/R(2 h/4 h)模型。实时荧光定量PCR(qRT-PCR)检测miR-6216的表达水平;细胞计数试剂盒(CCK-8)检测细胞存活;平板形成实验检测细胞克隆形成能力;试剂盒检测细胞内丙二醛(MDA)含量和细胞培养液中乳酸脱氢酶(LDH)的活性;流式细胞术检测细胞凋亡;蛋白质印记(Western blot)检测细胞周期蛋白D1(Cyclin D1)、P21、B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)的表达水平。将miR-6216抑制物(anti-miR-6216)、miR-6216模拟物(miR-6216 mimics)分别转染H9C2,将缺氧/复氧处理后,采用上述方法检测以上指标变化。结果缺氧/复氧诱导后H9C2细胞miR-6216的表达水平显著升高,细胞存活率和克隆能力显著降低,MDA含量和LDH活性显著升高,Cyclin D1和Bcl-2的表达显著降低,P21和Bax的表达水平显著升高,细胞凋亡率显著升高(P<0.05)。抑制miR-6216可显著提高H9C2细胞存活率和克隆形成能力,降低MDA含量和LDH活性,促进Cyclin D1和Bcl-2的表达,抑制P21和Bax的表达,抑制缺氧复氧诱导的细胞凋亡(P<0.05)。过表达miR-6216可显著抑制细胞存活和克隆形成能力,升高MDA含量和LDH活性,抑制Cyclin D1和Bcl-2的表达,促进P21和Bax的表达,加剧缺氧复氧诱导的细胞凋亡(P<0.05)。结论miR-6216在缺氧/复氧诱导的心肌细胞中表达上调,抑制miR-6216表达可减轻缺氧/复氧诱导的心肌细胞损伤。Objective To investigate the effect of miR-6216 on hypoxia/reoxygenation(H/R)-induced cardiomyocyte injury.Methods Rat embryonic cardiomyocytes H9 C2 were cultured in vitro to construct a H/R(2 h/4 h)model.Real-time quantitative PCR(qRT-PCR)was used to detect the expression level of miR-6216.Cell counting kit(CCK-8)was used to detect cell survival.The plate colony formation assay was used to detect cell clone formation ability.The kit was used to detect intracellular malondialdehyde(MDA)content and activity of lactate dehydrogenase(LDH)in cell culture medium.Apoptosis was detected by flow cytometry.The expression level of cyclin D1(Cyclin D1),P21,B cell lymphoma/leukemia-2(Bcl-2)and Bcl-2 related X protein(Bax)were detected by Western blot.The miR-6216 inhibitor(anti-miR-6216)and the miR-6216 mimics(miR-6216 mimics)were transfected into H9 C2,respectively.After the hypoxia/reoxygenation treatment,the above indicators were detected by the above method.Results The expression of miR-6216 in H9 C2 cells was significantly increased after hypoxia/reoxygenation induction,the cell viability and cloning ability were significantly decreased,MDA content and LDH activity were significantly increased,and the expression of Cyclin D1 and Bcl-2 was significantly decreased,and the expression levels of P21 and Bax were significantly increased,and the apoptosis rate was significantly increased(P<0.05).Inhibition of miR-6216 significantly increased H9 C2 cell survival and clonality,decreased MDA content and LDH activity,promoted Cyclin D1 and Bcl-2 expression,inhibited P21 and Bax expression,and inhibited hypoxia-reoxygenation-induced apoptosis(P<0.05).Overexpression of miR-6216 significantly inhibited cell survival and clonality,increased MDA content and LDH activity,inhibited the expression of Cyclin D1 and Bcl-2,promoted the expression of P21 and Bax,and aggravated hypoxia/reoxygenation-induced apoptosis(P<0.05).Conclusion miR-6216 is up-regulated in hypoxia/reoxygenation-induced cardiomyocytes,and inhibition of miR-6216 can atte

关 键 词：miR-6216 缺氧/复氧 心肌细胞损伤 凋亡 心肌梗死 

分 类 号：R542.22[医药卫生—心血管疾病]