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

魏飞宇; 范洁; 高田; 王礼琳

doi:10.13201/j.issn.1001-1439.2020.06.007

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

- 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)
基金项目:
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））

详细信息

通讯作者: 王礼琳,E-mail:kmwanglin@163.com

中图分类号: R541.75

收稿日期: 2020-03-18

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

- 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

More Information

Corresponding author: WANG Lilin, E-mail: kmwanglin@163.com

Received Date: 18 March 2020

摘要

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。
- 心房颤动 /
- miR-199-3p /
- 成纤维细胞增殖 /
- 结构重构 /
- SP1
Abstract: Objective: To investigate the relationship between miR-199-3p 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-199-3p.The diagnostic value of miR-199-3p in atrial fibrillation was analyzed by ROC.Pearson correlation was used to analyze the correlation between miR-199-3p 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-199-3p were determined by double luciferase reporter assay.Result: The expression level of miR-199-3p 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-199-3p 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-199-3p 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-199-3p 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-199-3p promoted fibroblast proliferation and fibrosis, but overexpression of miR-199-3p inhibited cell proliferation and fibrosis (P<0.05).At last, luciferase reporter assay confirmed that SP1 was the direct target gene of miR-199-3p.Conclusion: miR-199-3p is negatively correlated with left atrial fibrosis, and miR-199-3p negatively regulates fibroblast proliferation and fibrosis by targeting SP1 which contributes to atrial structural remodeling in atrial fibrillation.Therefore, miR-199-3p is expected to be a potential non-invasive diagnostic biomarker for atrial fibrillation and can also be a predictive biomarker of fibrosis.
- atrial fibrillation /
- miR-199-3p /
- fibroblast proliferation /
- structural remodeling /
- SP1

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