|
|||||
|
|
| 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 | |
| 引用本文： | 李海文,杨志明,高福萍,张瑞琴,柴蝉娟. 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[J]. 中国医学科学院学报, 2020, 42(5): 585-590. DOI: 10.3881/j.issn.1000-503X.11990 |
| 作者姓名： | 李海文 杨志明 高福萍 张瑞琴 柴蝉娟 |
| 作者单位： | 1.山西医科大学第二医院心内科,太原 0300012 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 |
| 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=... |
| 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 |
| 收稿时间： | 2019-09-10 |
Silencing miRNA210 Alleviates Myocardial Cell Damage in Myocarditis by Hypoxia Inducible Factor-vascular Endothelial Growth Factor Pathway |
|
| LI Haiwen,YANG Zhiming,GAO Fuping,ZHANG Ruiqin,CHAI Chanjuan. Silencing miRNA210 Alleviates Myocardial Cell Damage in Myocarditis by Hypoxia Inducible Factor-vascular Endothelial Growth Factor Pathway[J]. Acta Academiae Medicinae Sinicae, 2020, 42(5): 585-590. DOI: 10.3881/j.issn.1000-503X.11990 | |
| Authors: | LI Haiwen YANG Zhiming GAO Fuping ZHANG Ruiqin CHAI Chanjuan |
| Affiliation: | 1.Department of Cardiology,the Second Hospital of Shanxi Medical University,Taiyuan 030001,China2 Nanobiological Effects and Safety Laboratory,Institute of High Energy Physics,Chinese Academy of Sciences,Beijing 100049,China |
| Abstract: | 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)-α and interleukin(IL)-1β after 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-α and 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-1 β(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-1 β(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-1 β(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,P<0.001).Western blot analysis showed that,compared with the control group,LPS significantly down-regulated the expression of bcl-2(t=8.346,P=0.001)and increased the expressions of bax(t=12.890,P<0.001)and caspase-3(t=4.331,P=0.012).Compared with LPS group,the expression of bcl-2(t=6.074,P<0.001)was significantly decreased,and the expressions of bax(t=5.376,P=0.022)and caspase-3(t=5.859,P=0.004)were increased after miRNA210 mimic.After silencing miRNA210,the expression of bcl-2 significantly increased(t=3.873,P=0.017),the expressions of bax(t=5.205,P=0.006)and caspase-3(t=2.800,P=0.040)significantly decreased.Compared with the control group,the expressions of HIF1(t =10.380,P=0.001)and VEGF(t=4.973,P=0.008)were significantly up-regulated in LPS group.Compared with LPS group,the expressions of HIF1(t=8.952,P<0.001)and VEGF(t=11.203,P=0.001)were significantly up-regulated after miRNA210 mimic was applied,while HIF1(t=3.893,P=0.017)and VEGF expression(t=3.181,P=0.033)were decreased after miRNA210 was silenced.Compared with LPS group,the expressions of bax(t= 4.899,P=0.008),HIF1(t=2.833,P=0.047),caspase-3(t=2.877,P=0.045),and VEGF(t= 2.994, P=0.040)were significantly decreased,and the expression of bcl-2 was increased(t=3.392,P=0.017).Conclusion Silencing miRNA210 can attenuate LPS-induced cardiomyocyte injury through HIF1-VEGF-mediated apoptotic pathway. |
| Keywords: | miRNA210 myocarditis apoptotic pathway hypoxia inducible factor 1-vascular endothelial growth factor pathway |
| 点击此处可从《中国医学科学院学报》浏览原始摘要信息 | |
| 点击此处可从《中国医学科学院学报》下载全文 | |