摘要
目的探讨hsa_circ_0001946、hsa-miR-7-5p和人聚腺苷二磷酸核糖聚合酶1(PARP-1)在冠状动脉粥样硬化性心脏病(CHD)患者及内皮细胞功能障碍中的表达变化及意义。方法基于基因芯片初筛(n=5)和靶点预测,选择hsa_circ_0001946及其潜在的下游靶点hsa-miR-7-5p和PARP1做进一步的病例对照样本验证(n=30)。体外培养原代人脐静脉内皮细胞(HUVECs)和细胞系(EA.hy926),采用不同质量浓度的氧化低密度脂蛋白(oxLDL,0、1、2.5、5、10、25和50μg/mL)诱导内皮细胞功能障碍,采用Cell Counting Kit-8(CCK8)法检测细胞增殖活力,采用乳酸脱氢酶(LDH)试剂盒测定LDH活性,采用实时荧光定量聚合酶链反应(qRT-PCR)检测细胞间黏附分子-1(ICAM-1)、转化生长因子-β1(TGF-β1)、hsa_circ_0001946、hsa-miR-7-5p和PARP1的表达情况。结果CHD患者的hsa_circ_0001946表达上调,且与hsa-miR-7-5p呈负相关、与PARP1呈正相关(均P<0.05)。oxLDL可降低HUVECs和EA.hy926的细胞活力(其中HUVECs先升高后降低,P<0.05),可升高LDH活性、ICAM-1和TGF-β1的表达(P<0.05)。在低浓度oxLDL诱导的HUVECs和EA.hy926功能障碍中,hsa_circ_0001946和hsa-miR-7-5p表达均上调(P<0.05),PARP1表达下调(P<0.05)。结论Hsa_circ_0001946、hsa-miR-7-5p和PARP1在CHD患者和低浓度oxLDL导致的HUVECs和EA.hy926功能障碍中表达均异常,可能与CHD发生、发展密切相关的内皮细胞障碍有关。
Objective To explore the expression changes and significance of hsa_circ_0001946,hsa-miR-7-5p and poly ADP-ribose polymerase 1(PARP1)in patients with coronary atherosclerotic heart disease(CHD)and endothelial cell dysfunction.Methods Based on gene chip preliminary screening(n=5)and target prediction,hsa_circ_0001946and its potential downstream targets hsa-miR-7-5p and PARP1were selected to do further case-control sample validation(n=30).Primary human umbilical vein endothelial cells(HUVECs)and cell lines(EA.hy926)were cultured in vitro.Endothelial cell dysfunction was induced by oxidized low-density lipoprotein(oxLDL)at different concentrations(0,1,2.5,5,10,25and 50μg/mL)for 24h.Cell proliferation activity was measured by cell counting Kit-8(CCK8);lactate dehydrogenase(LDH)activity was measured by LDH kit;and intercellular adhesion molecule-1(ICAM-1),transforming growth factor-β1(TGF-β1),hsa_circ_0001946,hsa-miR-7-5p and PARP1expression were detected by quantitative real-time polymerase chain reaction(qRT-PCR).Results Hsa_circ_0001946expression up-regulated in patients with CHD,and was negatively correlated with hsa-miR-7-5p and positively correlated with PARP1(P<0.05).oxLDL significantly decreased the cell viability(HUVECs increased first and then decreased,P<0.05),and increased LDH activity,ICAM-1and TGF-β1(all P<0.05).Both hsa_circ_0001946and hsa-miR-7-5p expression were up-regulated(P<0.05)and PARP1expression was down-regulated(P<0.05)in HUVECs and EA.hy926dysfunction induced by low concentrations of oxLDL.Conclusion Hsa_circ_0001946,hsa-miR-7-5p and PARP1were all abnormally expressed in CHD patients and in HUVECs and EA.hy926dysfunction caused by low concentrations of oxLDL,which may be related to endothelial cell disorders closely related to the development of CHD.
作者
卢欣栎
黄淑娜
林杰
李爱娜
徐兴燕
杨乐
郭剑辉
吴思英
LU Xinyue;HUANG Shu'na;LIN Jie;LI Aina;XU Xingyan;YANG Le;GUO Jianhui;WU Siying(Department of Epidemiology and Health Statistics,School of Public Health,Fujian Medical University,Fuzhou 350122,China;Department of Clinical Research and Translation Center,The First Affiliated Hospital ofFujian Medical University,Fuzhou 350005,China;Department of Medical Administration,The Second Affiliated Hospital of Fujian Medical University,Quanzhou 362000,China;Department of Cardiology,The First Affiliated Hospital of Fujian Medical University,Fuzhou 350005,China)
出处
《福建医科大学学报》
2022年第6期516-523,共8页
Journal of Fujian Medical University
基金
国家自然科学基金项目(82173553)
福建省科技创新联合资金项目(2018Y9089)。
作者简介
通信作者:吴思英.Email:fmuwsy@163.com。
