沉默miRNA210可能通过低氧诱导因子-血管内皮生长因子途径减轻心肌炎时心肌细胞的损伤  

Silencing miRNA210 Alleviates Myocardial Cell Damage in Myocarditis by Hypoxia Inducible Factor-vascular Endothelial Growth Factor Pathway

作　　者：李海文[1] 杨志明[1] 高福萍 张瑞琴[1] 柴蝉娟[1] LI Haiwen;YANG Zhiming;GAO Fuping;ZHANG Ruiqin;CHAI Chanjuan(Department of Cardiology,the Second Hospital of Shanxi Medical University,Taiyuan 030001,China;Nanobiological Effects and Safety Laboratory,Institute of High Energy Physics,Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]山西医科大学第二医院心内科,太原030001 [2]中国科学院高能物理研究所,纳米生物效应与安全性研究室,北京100049

出　　处：《中国医学科学院学报》2020年第5期585-590,共6页Acta Academiae Medicinae Sinicae

摘　　要：目的研究miRNA210在脂多糖(LPS)诱导心肌炎时对大鼠原代心肌细胞的作用及其内在机制。方法采用CCK8法检测正常或LPS诱导时miRNA210对大鼠原代心肌细胞活力的影响;ELISA法检测在LPS诱导前提下,miRNA210处理后肿瘤坏死因子(TNF)-α与白细胞介素(IL)-1β的分泌情况;流式细胞凋亡法检测各干预组前后大鼠原代心肌细胞的凋亡情况;Western blot法检测凋亡相关蛋白bcl-2、bax、caspase-3和低氧诱导因子1(HIF1)-血管内皮生长因子(VEGF)的表达情况。结果CCK8检测结果显示,与对照组相比,miRNA210模拟物(t=0.000,P=1.000)和siRNA(t=0.686,P=0.500)干扰对大鼠原代心肌细胞的影响差异无统计学意义,LPS处理后大鼠原代心肌细胞的细胞活力明显降低(t=8.764,P<0.001);与LPS组相比,抑制miRNA210后大鼠原代心肌细胞活力明显升高(t=3.576,P=0.012)。ELISA检测结果显示,与对照组相比,LPS诱导后IL-1β(t=4.166,P=0.014)和TNF-α(t=6.309,P=0.003)表达显著上调;与LPS组相比,应用miRNA210模拟物后IL-1β(t=4.096,P=0.015)和TNF-α(t=4.424,P=0.011)表达显著升高,沉默miRNA210后IL-1β(t=4.287,P=0.012)和TNF-α(t=3.577,P=0.023)表达显著下调。流式细胞凋亡实验结果表明,与对照组相比,LPS诱导后的大鼠原代心肌细胞凋亡明显增加(t=32.780,P<0.001);与LPS组相比,应用miRNA210模拟物明显加重了LPS诱导的细胞凋亡(t=7.412,P=0.002),沉默miRNA210后的大鼠原代心肌细胞凋亡率明显降低(t=11.720,P<0.001)。Western blot检测结果表明,与对照组相比,LPS显著下调了bcl-2表达(t=8.346,P=0.001),上调了bax(t=12.890,P<0.001)和caspase-3的表达(t=4.331,P=0.012)。与LPS组相比,应用miRNA210模拟物后bcl-2(t=6.074,P=0.003)表达明显下降,bax(t=5.376,P=0.022)和caspase-3(t=5.859,P=0.004)表达明显升高;沉默miRNA210后bcl-2表达明显升高(t=3.873,P=0.017),bax(t=5.205,P=0.006)和caspase-3(t=2.800,P=0.040)表达明显下降。与对照组比,LPS组的HIF1(t=10.380,P=0.001)和VEGF(t=4.973,Objective To investigate the effect of miRNA210 on primary myocardial cells in lipopolysaccharide(LPS)-induced myocarditis.Methods CCK8 method was used to detect the effect of miRNA210 on the viability of primary myocardial cells in normal or LPS-induced myocarditis rats.ELISA was performed to detect the secretion of tumor necrosis factor(TNF)-αand interleukin(IL)-1βafter miRNA210 treatment.Flow cytometry was used to detect the apoptosis of primary myocardial cells before and after the intervention.Western blotting was used to detect the expression of TNF-αand IL-1β.The expression of apoptosis-related proteins bcl-2,bax,caspase-3,and hypoxia inducible factor 1(HIF1)-vascular endothelial growth factor(VEGF)were detected by Western blotting.Results CCK8 detection results showed that,compared with the control group,the effect of miRNA210 mimic(t=0.000,P=1.000)and siRNA(t=0.686,P=0.500)interference had no significant difference on primary rat cardiomyocytes.The viability of rat primary cardiomyocytes significantly decreased after LPS treatment(t=8.764,P<0.001);compared with LPS group,the viability of rat primary cardiomyocytes significantly increased after inhibition of miRNA210(t=3.576,P=0.012).ELISA showed that,compared with the control group,the expressions of IL-1β(t=4.166,P=0.014)and TNF-α(t=6.309,P=0.003)were significantly up-regulated after LPS induction;compared with the LPS group,the expressions of IL-1β(t=4.096,P=0.015)and TNF-α(t=4.424,P=0.011)were significantly increased after application of miRNA210 mimic.After silencing miRNA210,IL-1β(t=4.287,P=0.012)and TNF-α(t=3.577,P=0.023)were significantly down-regulated.Flow cytometry showed that,compared with the control group,the apoptosis of primary cardiomyocytes induced by LPS was significantly increased(t=32.780,P<0.001);compared with LPS group,the apoptosis induced by LPS was significantly aggravated by miRNA210 mimic(t=7.412,P=0.002),and the apoptosis rate of primary cardiomyocytes was significantly decreased after miRNA210 was silenced(t=11.720,

关 键 词：miRNA210 心肌炎 凋亡通路 低氧诱导因子1-血管内皮生长因子通路 

分 类 号：R542.2+1[医药卫生—心血管疾病]