微小RNA-133与微小RNA-208在急性心肌梗死患者中的免疫学作用
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摘要
目的研究微小RNA-133与微小RNA-208在心肌梗死患者中的免疫学作用。方法 qPCR检测健康对照组与急性心肌梗死(acute myocardial infarction,AMI)组患者不同时段血浆中miR-133与miR-208的相对表达量;转染特异性干扰物与模拟物改变心肌HCM细胞中miR-133与miR-208的表达量,流式细胞仪检测HCM凋亡;CCK-8检测HCM细胞增殖活力;Western blot检测miR-133与miR-208对下游相关免疫调节蛋白CCL2、CX3CL1、RGC32的蛋白表达量的影响。结果 0 h与24 h组急性心肌梗死组患者中miR-133与miR-208较健康对照组均明显上升,7 d后恢复至正常水平;miR-133与miR-208可以促进心肌HCM细胞凋亡;miR-133与miR-208会减弱心肌HCM细胞的增殖活力;miR-133与miR-208会降低免疫调节蛋白CCL2、CX3CL1、RGC32的表达量。结论 AMI患者血浆miR-133与miR-208水平有显著升高,miR-133与miR-208可以通过介导急性心梗患者免疫调节因子CCL2、CX3CL1与RGC32的表达上调,促进心肌HCM细胞的增殖,抑制心肌HCM细胞的凋亡,从而起到保护心肌细胞的作用。
This study was performed to investigate the immunological function of miR-133 and miR-208 increased in the acute myocardial infarction(AMI) patients, and to investigate the clinical relevance of miR-133 and miR-208 expression in AMI. The expression of miR-133 and miR-208 were detected by quantitative real time PCR(qRT-PCR) in blood of 138 AMI patients and 86 healthy controls; miRNA's specific siRNA and mimics were transfected to change the levels of miR-133 and miR-208 in HCM cell. CCK-8 assays was used to detect the cell proliferation, FCM flow cytometry was employed to detect apoptosis rate, while Western blot was applied to measure the protein levels of CCL2/CX3 CL1/RGC32. Data showed that the levels of miR-133 and miR-208 in AMI group were higher than those of healthy group in 0 h and 24 h, and returned to normal levels 7 days later; miR-133 and miR-208 could promote HCM cell apoptosis and inhibit the HCM cell proliferation, and could up-regulate the protein of immune-regulation genes(CCL2/CX3 CL1/RGC32). In conclusion, miR-133 and miR-208 can promote the cell proliferation and inhibit the apoptosis of HCM by up-regulating the activities of CCL2/CX3 CL1/RGC32,thus protect the patients' myocardial cells.
This study was performed to investigate the immunological function of miR-133 and miR-208 increased in the acute myocardial infarction(AMI) patients, and to investigate the clinical relevance of miR-133 and miR-208 expression in AMI. The expression of miR-133 and miR-208 were detected by quantitative real time PCR(qRT-PCR) in blood of 138 AMI patients and 86 healthy controls; miRNA's specific siRNA and mimics were transfected to change the levels of miR-133 and miR-208 in HCM cell. CCK-8 assays was used to detect the cell proliferation, FCM flow cytometry was employed to detect apoptosis rate, while Western blot was applied to measure the protein levels of CCL2/CX3 CL1/RGC32. Data showed that the levels of miR-133 and miR-208 in AMI group were higher than those of healthy group in 0 h and 24 h, and returned to normal levels 7 days later; miR-133 and miR-208 could promote HCM cell apoptosis and inhibit the HCM cell proliferation, and could up-regulate the protein of immune-regulation genes(CCL2/CX3 CL1/RGC32). In conclusion, miR-133 and miR-208 can promote the cell proliferation and inhibit the apoptosis of HCM by up-regulating the activities of CCL2/CX3 CL1/RGC32,thus protect the patients' myocardial cells.
引文
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[3]曹禹,张效林,刘丹,等.血浆中趋化因子CCL2水平与ST抬高型心肌梗死患者预后相关性研究[J].临床军医杂志,2017,45(6):561-565.
[4]廉铮,吕峰峰,王家旺.冠心病外周血中CX3CL1、CX3CR1的表达及临床意义[J].中国循证心血管医学杂志,2018,10(8):919-922.
[5]王珏.MicroRNA-24调控心肌梗死后纤维化过程的实验研究[D].北京:北京协和医学院,2012.
[6]Thygesen K,Alpert JS,Jaffe AS,et al.Third universal definition of myocardial infarction[J].Glob Heart,2012,7(4):275-295.
[7]徐振宇,杨立明,李广平,等.慢性心力衰竭患者血清miR-133a、miR-133b水平与心功能和心肌重构的关系[J].山东医药,2018,58(4):82-84.
[8]曾德银,刘茜.急性心肌梗死患者血清miR-208对血小板功能、炎症反应及心肌损伤程度的影响[J].海南医学院学报,2018,24(6):657-660.
[9]王瑾,马肖容,张王刚.CCK-8法在淋巴细胞增殖检测中最佳实验条件的筛选[J].中国医药导报,2018,15(23):13-16.
[10]Han M,Toli J,Abdellatif M.MicroRNAs in the cardiovascular system[J].Curr Opin Cardiol,2011,26(3):181-189.
[11]Abdellatif M.The role of microRNA-133 in cardiac hypertrophy uncovered[J].Circ Res,2010,106(1):16-18.
[12]Danowski N,Manthey I,Jakob HG,et al.Decreased expression of miR-133a but not of miR-1 is associated with signs of heart failure in patients undergoing coronary bypass surgery[J].Cardiology,2013,125(2):125-130.
[13]He B,Xiao J,Ren AJ,et al.Role of miR-1 and miR-133a in myocardial ischemic postconditioning[J].J Biomed Sci,2011,18:22.
[14]D'Alessandra Y,Devanna P,Limana F,et al.Circulating microRNAs are new and sensitive biomarkers of myocardial infarction[J].Eur Heart J,2010,31(22):2765-2773.
[15]Li C,Pei F,Zhu X,et al.Circulating microRNAs as novel and sensitive biomarkers of acute myocardial Infarction[J].Clin Biochem,2012,45(10/11):727-732.
[16]Nabel EG,Braunwald E.A tale of coronary artery disease and myocardial infarction[J].N Engl J Med,2012,366(1):54-63.
[17]王晨阳,刘海燕,曹海涛,等.心肌梗死发生和发展过程中的免疫机制研究[J].解放军医药杂志,2014,26(11):21-23.
[18]Nahrendorf M,Swirski FK,Aikawa E,et al.The healing myocardium sequentially mobilizes two monocyte subsets with divergent and complementary functions[J].J Exp Med,2007,204(12):3037-3047.
[19]Dewald O,Zymek P,Winkelmann K,et al.CCL2/monocyte chemoattractant protein-1 regulates inflammatory responses critical to healing myocardial infarcts[J].Circ Res,2005,96(8):881-889.