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欄目介紹 彤心飛傳是由天津醫(yī)科大學第二醫(yī)院心臟科劉彤教授和鄭州大學附屬洛陽中心醫(yī)院心內(nèi)科谷云飛共同推出的一檔心血管前沿文獻速讀欄目。欄目的理念是“專注 堅持 積累”，每期精選國際頂級期刊心血管領(lǐng)域的最新文獻進行深度呈現(xiàn)，以期讓國內(nèi)相關(guān)專業(yè)人員可以同步跟蹤學科進展，了解前沿動態(tài)資訊，獲取最新研究結(jié)論。 
心房顫動（AF）是臨床最常見的房性心律失常，明顯增加栓塞和心力衰竭發(fā)生的風險[1,2]。指南推薦癥狀性房顫首選導管射頻消融治療[3]。然而，雖然陣發(fā)性房顫療效較好，但持續(xù)性房顫術(shù)后長期復發(fā)率仍然較高。心房重構(gòu)是房顫發(fā)生的主要病理機制，心房纖維化是心房重構(gòu)的重要標志，心房纖維化的嚴重程度與房顫的預后有關(guān)[4]。改善左房纖維化基質(zhì)可改善房顫的預后[5]。因此、尋找心房纖維化的早期生物標志物并探討房顫的發(fā)病機制對改善預后至關(guān)重要。miRNA是一種長度在18-22堿基的非編碼RNA分子，目前研究證實其參與調(diào)控多種心血管疾病的發(fā)生發(fā)展。且循環(huán)中的miRNAs可作為各種疾病的診斷生物學標志物[6,7]。 最近，云南省第一人民醫(yī)院范潔課題組在Journal of Cardiology發(fā)表原創(chuàng)研究，題目為miR-425-5p is negatively associated with atrial fibrosis and promotes atrial remodeling by targeting CREB1 in atrial fibrillation，通過使用高通量測序來篩選房顫患者及非房顫患者血液中差異表達的miRNAs，并分析其與左房纖維化的關(guān)系及射頻消融術(shù)后的表達變化。同時在細胞和動物模型中驗證miR-425-5p在房顫心房重構(gòu)中的作用及作用分子機制。 首先通過高通量測序發(fā)現(xiàn)mIR-425-5p在房顫患者外周血中表達下降，并進一步通過qRT-PCR在80例房顫患者外周血及外科手術(shù)房顫患者心房組織標本驗證其表達下降，更為重要的是，房顫患者左房血液標本中miR-425-5p表達低于外周血，進一步提示miR-425-5p的異常表達可能與房顫發(fā)生發(fā)展密切相關(guān)。 其次分析miR-425-5p在不同類型房顫的表達變化，結(jié)果發(fā)現(xiàn)miR-425-5p在持續(xù)性心房顫動患者中表達低于陣發(fā)性房顫，同時對接受房顫導管射頻消融術(shù)的持續(xù)性房顫術(shù)中標測左房低電壓區(qū)，分析miR-425-5p表達與左房低電壓的關(guān)系，結(jié)果顯示miR-425-5p與持續(xù)性房顫患者左房低電壓區(qū)呈負相關(guān)，提示miR-425-5p與左房纖維化相關(guān)。更為重要的是，導管消融可逆轉(zhuǎn)血漿中下降的miR-425-5p表達。 最后通過構(gòu)建房顫細胞模型與動物模型，利用分子生物學技術(shù)，證實下調(diào)miR-425-5p表達可促進心房成纖維細胞增殖，而過表達miR-425-5p表達可改善房顫心房重構(gòu)。進一步利用靶基因預測軟件，熒光素酶報告基因?qū)嶒?，免疫印跡等實驗，證實miR-425-5p通過直接靶向調(diào)控CREB1參與調(diào)控上述生物學功能。 Figure1: The expression level of miRNAs in plasma in two dependent cohorts by miRNA sequencing and qRT-PCR. A: Heatmap of miRNA expression data from plasma samples of patients with AF(n=3) and healthy controls (n=3). Relative miRNA expression was depicted according to the color scale shown on the botton. Red indicated upregulation; green indicated downregulation, Ctl indicated healthy control samples. B-C: Validation of miRNA sequencing data by qRT-PCR in patients with AF(n=3) and healthy controls (n=3), **P<0.01 vs control.D-F: Validation of 2 downregulation miRNAs and 1 upregulation miRNAexpression in an independent cohort with 80 AF patients and 80 healthy control by qRT-PCR, ***P<0.001. The vertical axis was ΔCt. whereby a lower ΔCt value mean a higher expression of miRNA.
Figure2: miR-425-5p was associated with left atrial fibrosis. A: The miR-425-5p expression level in persistent AF and paroxysmal AF, paro AF= paroxysmal AF, pers AF= persistent AF, The vertical axis was ΔCt. whereby a lower ΔCt value mean a higher expression of miRNA. B: The correlation between plasma miR-425-5p and the extent of left atrial low voltage area, all valve were log-transformed to normalize their distribution before statistical analysis, the extent of left atrial low voltage was used to evaluate left fibrosis. C-E: The representative image of the left atrial low voltage area(LVA), the red color represented the left atrial low area which reflect the severity of left atrial fibrosis. F: Relative miR-425-5p expression according to LVA group(group 1: LVAs <10%; group 2: LVAs 10% to 20%; group 3: LVAs>20%, *P<0.05, ***P<0.001).
Figure3: The miR-425-5p expression level in atrial tissue in AF patients and mouse AF model. A: The expression level of miR-425-5p in AF patients(n=6); B: The expression level of miR-425-5p in mouse AF model(n=12); C: The expression level of miR-425-5p in left and right atrium tissue in AF model and sham, **P<0.01.
Figure4: The plasma miR-425-5P expression level in the pre-RFCA and post-RFCA. A: The plasma level of miR-425-5p in the control group (n=20) and pre-RFCA (n=20), post-RFCA (n=20), **P<0.01 vs control group, ##P<0.01 vs pre-RFCA. B: The expression change of miR-425-5p in each AF patients in pre-RFCA and post-RFCA.
Figure5: Effect of miR-425-5p on atrial remodeling. A: The miR-425-5p expression level in fibroblast cell transfected with miR-425-5p inhibitor. B: Fibroblast cell proliferation was detected with CCK-8. C: The proliferation markers of Ki67 and Cyclin D. D: The expression level of fibrosis biomarkers. E: The miR-425-5p expression level in mouse atrial tissue injected with miR-425-5p mimics. F: The AF incidence rate. G: Typical representative Masson’s trichrome staining. H: The statistical histogram, ***P<0.001, **P<0.01
Figure6: CREB1 was the direct target gene of miR-425-5p. A: The CREB1 expression was upregulated in atrial tissue in AF patients and mouse AF model. B: The CREB1 expression was negatively with miR-425-5p in AF patients by Pearson’s correlation coefficients. C-D: Inhibition of CREB1 expression could reverse the phenotype of downregulation of miR-425-5p to fibroblast cell proliferation and fibrosis. cell proliferation was detected with CCK-8. E: The potential binding site of miR-425-5p with CREB1 3’-URT was indicated with TargetScan 7.2 database. F-G: The CREB1 protein expression level in atrial fibroblast cell transfected with miR-NC and miR-425-5p inhibitor, GAPDH was used as loading control. H: Luciferase assay confirmed that CREB1 was a target of miR-425-5p, **P<0.01,*P<0.05.綜述結(jié)果表明，miR-425-5p在AF患者血漿和心房組織中表達下調(diào)，miR-425-5p下調(diào)可作為心房纖維化的無創(chuàng)生物標志物，進而通過靶向調(diào)控CREB1表達參與調(diào)控房顫心房重構(gòu)。進一步揭示了miRNAs在房顫發(fā)生發(fā)展中的作用，miR-425-5p有望成為未來房顫干預的分子靶點及預后的分子標志物。1. Odutayo A, et al. Atrial fibrillation and risks of cardiovascular disease, renal disease, and death: systematic review and meta-analysis. BMJ 2016;354:i4482.2. Bekwelem W, et al. Extracranial systemic embolic events in patients with nonvalvular atrial fibrillation. Circulation 2015;132:796-803.3. Tilz RR, et al. Ten-year clinical outcome after circumferential pulmonary vein isolation utilizing the Hamburg approach in patients with symptomatic drug-refractory paroxysmal atrial fibrillation. Circ Arrhythm Electrophysiol 2018;11:e005250.4. Thanigaimani S, et al. Molecular mechanisms of atrial fibrosis: implications for the clinic. Expert Rev Cardiovasc Ther 2017;15:247-56.5 Yang B, et al. STABLE-SR (Electrophysiological Substrate Ablation in the Left Atrium During Sinus Rhythm) for the Treatment of Nonparoxysmal Atrial Fibrillation: A prospective, multicenter randomized clinical trial. Circ Arrhythm Electrophysiol 2017;10:e005405.6. Fan PC, et al. A circulating miRNA signature for early diagnosis of acute kidney injury following acute myocardial infarction. J Transl Med 2019;17:139.7. Aghabozorgi AS, et al. Circulating exosomal miRNAs in cardiovascular disease pathogenesis: New emerging hopes. J Cell Physiol 2019;234:21796-809.范潔，云南省第一人民醫(yī)院心內(nèi)科副主任、國家衛(wèi)計委心律失常介入培訓基地負責人，博士生導師、二級教授， 現(xiàn)任中華醫(yī)學會心電生理和起搏分會委員，女醫(yī)師聯(lián)盟的常委，中國醫(yī)師協(xié)會心律失常學專業(yè)委員會常委，云南省醫(yī)學會心電生理和起搏學會的副主任委員。先后發(fā)表學術(shù)論文40余篇，主編專著一本。獲云南省科技進步一等獎，獲得國家基金3項、省基金面上項目資助3項。
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