miR-199-3p通过靶向调控SP1促进成纤维细胞增殖对心房颤动心房重构的影响  

The effect of miR-199-3p on fibroblasts proliferation to promote atrial structural remodeling by targeting SP1 in atrial fibrillation

作　　者：魏飞宇 范洁[1] 高田[1] 王礼琳[1] WEI Feiyu;FAN Jie;GAO Tian;WANG Lilin(Department of Cardiology,Yunnan Arrhythmia Research Center,The First People's Hospital of Yunnan Province,Affiliated Hospital of Kunming University of Science and Technology,Kunming,650032,China)

机构地区：[1]云南省第一人民医院,昆明理工大学附属医院心血管内科,云南省心律失常诊疗中心,昆明650032

出　　处：《临床心血管病杂志》2020年第6期523-530,共8页Journal of Clinical Cardiology

基　　金：云南省科技厅-昆明医科大学应用基础研究联合专项面上项目(No:2019FE001(-291))。

摘　　要：目的:探讨miR-199-3p与心房纤维化的关系及其对心房成纤维细胞增殖的影响。方法:连续入选2018年11月-2019年11月在云南省第一人民医院心内科住院的心房颤动(AF)患者100例,以基线资料相匹配的非AF患者50例作为对照,利用qRT-PCR检测2组患者外周血中miR-199-3p的表达差异;利用受试者工作特征曲线(ROC)分析miR-199-3p对AF的诊断价值;采用Pearson相关分析miR-199-3p和左房纤维化的相关性;利用CCK-8测定miR-199-3p对成纤维细胞增殖的影响,qRT-PCR检测细胞周期调控因子Ki67和Cyclin D1及CollagenⅠ和CollagenⅢ表达变化;最后利用双荧光素酶报告基因实验确定miR-199-3p的靶基因。结果:miR-199-3p在AF患者外周血[(0.38±0.31)∶(1.25±0.89),P<0.01]和心房组织中[(0.48±0.03)∶(1.00±0.12),P<0.01]表达都下调,且在持续性AF患者中表达低于阵发性AF[(0.42±0.20)∶(1.08±0.48),P<0.01];ROC曲线分析miR-199-3p表达下降鉴别AF的曲线下面积(AUC)为0.90,其敏感性为81%,特异性为86%,鉴别持续性AF的AUC为0.91,其敏感性为98%,特异性为73%;Pearson相关分析显示miR-199-3p表达水平和左房纤维化程度呈负相关(r=-0.863,P<0.01);与对照组相比,沉默miR-199-3p表达可促进成纤维细胞增殖和纤维化,但过表达miR-199-3p可抑制细胞增殖和纤维化(P<0.05);荧光素酶报告基因实验和Western blot证实SP1是miR-199-3p的直接靶基因。结论:AF患者中miR-199-3p和左房纤维化呈负相关,且miR-199-3p通过靶向SP1的表达负性调控成纤维细胞增殖和纤维化,进而参与AF心房重构。因此,miR-199-3p有望成为临床AF潜在的非侵入性诊断标志物和纤维化的预测标志物。Objective:To investigate the relationship between miR-199-3 p and atrial fibrosis and its effect on cardiac fibroblast proliferation.Method:One hundred patients with atrial fibrillation hospitalized in our centre from November 2018 to November 2019 were selected consecutively,and 50 non-atrial fibrillation patients matched with baseline data were selected as the control group.qRT-PCR was used to detect the expression of miR-199-3 p.The diagnostic value of miR-199-3 p in atrial fibrillation was analyzed by ROC.Pearson correlation was used to analyze the correlation between miR-199-3 p and left atrial fibrosis.Cell proliferation activity was determined by CCK-8,qRT-PCR was used to detect the mRNA expression level of Ki67,Cyclin D1,Collagen I and Collagen III.Finally,the target genes of miR-199-3 p were determined by double luciferase reporter assay.Result:The expression level of miR-199-3 p in peripheral blood[(0.38±0.31)∶(1.25±0.89),P<0.01]and atrial tissues[(0.48±0.03)∶(1.00±0.12),P<0.01]of patients with atrial fibrillation was down-regulated compared with control,and the expression level in persistent atrial fibrillation was lower than that in paroxysmal atrial fibrillation[(0.42±0.20)∶(1.08±0.48),P<0.01].The miR-199-3 p ROC curve analysis of AF showed that the area under the curve was 0.90 with a sensitivity of 81%and a specificity of 76%.Moreover,the miR-199-3 p ROC curve analysis of persistent AF showed that the area under the curve was 0.91 with a sensitivity of 98%and a specificity of 73%.Pearson correlation analysis showed that miR-199-3 p was negatively correlated with the degree of left atrial fibrosis(r=-0.863,P<0.01).Compared with the control group,silencing the expression of miR-199-3 p promoted fibroblast proliferation and fibrosis,but overexpression of miR-199-3 p inhibited cell proliferation and fibrosis(P<0.05).At last,luciferase reporter assay confirmed that SP1 was the direct target gene of miR-199-3 p.Conclusion:miR-199-3 p is negatively correlated with left atrial fibrosis,and miR-

关 键 词：心房颤动 miR-199-3p 成纤维细胞增殖 结构重构 SP1 

分 类 号：R541.75[医药卫生—心血管疾病]