摘要
[目的]探讨血浆miR-1228-5p、miR-34a-5p、miR-192-5p和miR-30a-3p水平与早发冠心病(PCAD)的相关性及其对PCAD的初筛价值。[方法]根据纳入标准及排除标准,纳入6例明确诊断的PCAD患者作为PCAD组,纳入6例健康受试者作为对照组,收集PCAD组和对照组血液,提取血清样本并保存,使用DNBseq平台检测两组血清中miRNA水平,筛选差异水平显著的miRNA。根据纳入标准及排除标准,收集78例PCAD患者、75例晚发冠心病患者和69例健康对照者的血液并对筛选的miRNA进行实时荧光定量PCR验证。分析PCAD患者冠状动脉造影报告,采用Gensini评分评估冠状动脉病变的严重程度。Spearman相关性检验分析有关miRNA水平与冠状动脉狭窄程度的相关性。ROC曲线分析血浆miR-1228-5p、miR-34a-5p、miR-192-5p及miR-30a-3p水平对PCAD的诊断价值,多因素Logistic回归分析PCAD发生的影响因素。[结果]DNBseq平台分析显示,差异表达miRNA 33个,其中上调miRNA 17个,下调miRNA 16个,差异水平最为显著的5个miRNA分别为miR-1228-5p、miR-34a-5p、miR-192-5p、miR-424-3p和miR-30a-3p;实时荧光定量PCR结果显示,与对照组相比,PCAD患者血浆miR-1228-5p升高1.7倍,miR-34a-5p升高1.4倍,miR-192-5p升高0.7倍,miR-30a-3p升高2.5倍(P<0.05),两组间血浆miR-424-3p水平差异无统计学意义(P>0.05);血浆miR-1228-5p和miR-34a-5p水平与PCAD患者冠状动脉狭窄程度均呈正相关(r=0.307,P=0.004;r=0.238,P=0.036);ROC曲线分析显示,miR-1228-5p、miR-34a-5p、miR-192-5p和miR-30a-3p诊断PCAD的ROC曲线下面积分别为0.903、0.832、0.731及0.798,其联合诊断PCAD的ROC曲线下面积为0.990,95%CI为0.976~1.000。[结论]PCAD患者血浆miR-1228-5p、miR-34a-5p、miR-192-5p和miR-30a-3p水平显著升高,其联合检测诊断PCAD具有较高的准确性,有望成为初筛PCAD的新型生物标志物。
Aim To investigate the correlation between plasma miR-1228-5p,miR-34a-5p,miR-192-5p and miR-30a-3p levels and premature coronary artery disease(PCAD)and their early screening value as biomarkers.Methods According to inclusion criteria and exclusion criteria,6 patients with clearly diagnosed PCAD were included as PCAD group and 6 heathy subjects as control group.Blood was collected from the PCAD group and the control group,and serum samples were extracted and stored.The miRNA levels in the serum of the two groups were detected using DNBseq platform,and the miRNA with significant differences were screened.According to inclusion and exclusion criteria,blood from 78 patients with PCAD,75 patients with late-onset coronary artery disease and 69 healthy subjects were collected and the selected miRNAs were verified by qPCR.Coronary angiography reports of patients with PCAD were analyzed,and the severity of coronary artery disease was evaluated by Gensini score.Spearman correlation test was used to analyze the correlation between the degree of coronary stenosis and the miRNA.ROC curve was used to analyze the diagnostic value of plasma levels of miR-1228-5p,miR-34a-5p,miR-192-5p and miR-30a-3p for PCAD,and multivariate Logistic regression was used to analyze the influencing factors of PCAD.Results DNBseq platform analysis showed 33 differentially expressed miRNAs,including 17 up-regulated miRNAs and 16 down-regulated miRNAs.The 5 miRNAs with the most significant difference levels were miR-1228-5p,miR-34a-5p,miR-192-5p,miR-424-3p and miR-30a-3p.qPCR results showed that compared with the control group,plasma miR-1228-5p,miR-34a-5p,miR-192-5p and miR-30a-3p levels were increased by 1.7 times,1.4 times,0.7 times and 2.5 times in patients with PCAD(P<0.05).There was no significant difference in miR-424-3p level between the two groups(P>0.05).Spearman correlation analysis showed that plasma levels of miR-1228-5p and miR-34a-5p were positively correlated with the degree of coronary stenosis in patients with PCAD(r=0.307,P=0.004;r=0.238,P=0.036).ROC curve analysis showed that the areas under ROC curve for the diagnosis of PCAD by miR-1228-5p,miR-34a-5p,miR-192-5p and miR-30a-3p were 0.903,0.832,0.731 and 0.798,respectively.The area under ROC curve of the combined diagnosis was 0.990,and 95%CI was 0.970~1.000.Conclusions The plasma levels of miR-1228-5p,miR-34a-5p,miR-192-5p and miR-30a-3p in patients with PCAD were significantly increased.The combined detection showed high accuracy in the diagnosis of PCAD,which is expected to be new biomarkers for preliminary screening of PCAD.
作者
田开疆
周坤
桂丹丹
罗雯
欧阳劭
马小峰
刘欢
胡恒境
姜志胜
TIAN Kaijiang;ZHOU Kun;GUI Dandan;LUO Wen;OU-YANG Shao;MA Xiaofeng;LIU Huan;HU Hengjing;JIANG Zhisheng(Institute of Cardiovascular Disease,University of South China&Key Laboratory for Arteriosclerology of Hunan Province&International Joint Laboratory for Arteriosclerotic Disease Research of Hunan Province,Hengyang,Hunan 421001,China;Department of Cardiology,the Second Affiliated Hospital of University of South China,Hengyang,Hunan 421001,China;Department of Cardiology,the Affiliated Nanhua Hospital of University of South China,Hengyang,Hunan 421002,China;Department of Cardiology,the First Affiliated Hospital of University of South China,Hengyang,Hunan 421001,China)
出处
《中国动脉硬化杂志》
CAS
2022年第8期681-690,共10页
Chinese Journal of Arteriosclerosis
基金
国家自然科学基金项目(91839103、81670429和81470435)
湖南省重点研发计划(2020SK2105)。
作者简介
田开疆,硕士研究生,研究方向为动脉粥样硬化病因发病学与防治基础,E-mail:842549485@qq.com。;通信作者:姜志胜,教授,博士研究生导师,研究方向为动脉粥样硬化及其发病机制,E-mail:zsjiang2016@163.com。
