NLRC3在血管内皮细胞中的功能初探及在冠心病中的诊断价值  

Function of NLRC3 in endothelial cells and its diagnostic significance in coronary artery disease

作　　者：古晓东 翁锐强 赵俊丽 李霞 刘苏东 GU Xiaodong;WENG Ruiqiang;ZHAO Junli;LI Xia;LIU Sudong(Meizhou Clinical Institute,Shantou University Medical College,Meizhou 514000,China;Institute of Basic Medical Sci-ences,Meizhou People's Hospital(Huangtang Hospital),Meizhou 514000,China;State Key Laboratory of Neurology and Oncology Drug Development,Nanjing 210000,China)

机构地区：[1]汕头大学医学院梅州临床学院,广东梅州514000 [2]梅州市人民医院(黄塘医院)基础医学研究所,广东梅州514000 [3]神经与肿瘤药物研发全国重点实验室,江苏南京210000

出　　处：《中国病理生理杂志》2025年第4期661-668,共8页Chinese Journal of Pathophysiology

基　　金：广东省基础与应用基础研究基金企业联合基金面上项目(No.2024A1515220103);广东省基础与应用基础研究基金面上项目(No.2024A1515010809);梅州市社会发展科技计划项目(No.2023B04)。

摘　　要：目的:探讨NLR家族含CARD域蛋白3(NLR family CARD domain containing 3,NLRC3)在血管内皮细胞中的功能及在冠心病中的诊断价值。方法:取20只8周龄雄性ApoE^(-/-)C57BL/6小鼠,分为实验组及对照组(各10只),实验组采用高脂饮食喂养8周诱导动脉粥样硬化(atherosclerosis,AS),对照组采用普通饮食喂养;采用RT-qPCR及免疫荧光检测主动脉NLRC3的表达;采用白细胞介素1β(interleukin-1β,IL-1β;10μg/L)刺激人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs),检测NLRC3的表达变化;采用慢病毒载体或质粒载体在内皮细胞中过表达或敲减NLRC3的表达并用IL-1β诱导炎症,采用RT-qPCR和ELISA检测NLRC3对内皮细胞炎症的影响,采用Western blot和免疫荧光检测NLRC3对内皮细胞NF-κB信号通路的调控;采用ELISA检测冠心病患者及对照患者血浆NLRC3的表达水平,通过受试者工作特征曲线评估检测NLRC3在冠心病诊断中的价值。结果:AS小鼠主动脉中可观察到明显的粥样斑块病变;NLRC3在AS小鼠主动脉弓的表达较对照组小鼠下调;NLRC3在IL-1β刺激的HUVECs中显著下调,并呈现时间依赖性(P<0.01)和剂量依赖性(P<0.01);过表达NLRC3显著抑制IL-1β刺激的内皮细胞IL-6、IL-8、单核细胞趋化蛋白1(monocyte chemoatbactant protein-1,MCP-1)、p-p65及p-IκBα的表达(P<0.01),而敲减NLRC3则增加内皮细胞IL-6、IL-8、MCP-1、p-p65及p-IκBα的表达(P<0.01);NLRC3在冠心病患者血浆中的表达水平显著低于对照组,其诊断冠心病的曲线下面积为0.851,95%CI为0.785~0.918(P<0.01),当临界值为1.605μg/L时,诊断灵敏度为93.8%,特异度为71.3%。结论:NLRC3可能通过抑制NF-κB通路调节内皮炎症影响AS的发展,并可作为冠心病诊断的潜在生物标志物。AIM:To investigate the role of NLR family CARD domain containing 3(NLRC3)in endothelial cells and evaluate its diagnostic value in coronary artery disease.METHODS:Twenty male ApoE^(-/-)C57BL/6 mice,aged eight weeks,were randomly assigned into two groups:an experimental group and a control group,with each group comprising ten mice.The experimental group was subjected to a high-fat diet for 8 weeks to induce atherosclerosis(AS),whereas the control group was maintained on a standard diet.The expression of NLRC3 in the aorta was evaluated using RT-qPCR and immunofluorescence techniques.Additionally,human umbilical vein endothelial cells(HUVECs)were exposed to interleukin-1β(IL-1β)to investigate the expression levels of NLRC3.Lentiviral vectors or plasmid vectors were employed to either overexpress or knock down NLRC3 in endothelial cells,and subsequently subjected to inflammation induced by IL-1β.The RT-qPCR and ELISA were employed to assess the impact of NLRC3 on inflammation in endothelial cells.Western blot and immunofluorescence techniques were utilized to investigate the modulation of the NF-κB signaling pathway in endothelial cells by NLRC3.Plasma NLRC3 levels in coronary artery disease patients and healthy controls were measured using ELISA,and its diagnostic potential was assessed through ROC curve analysis.RESULTS:In AS mice,distinct plaque lesions were observed in the aorta,accompanied by a significantly reduced expression of NLRC3 in the aortic arch relative to the control group.Expression of NLRC3 exhibited a significant down-regulation in IL-1β-stimulated HUVECs,demonstrating both time-dependent and dose-dependent effects(P<0.01).Overexpression of NLRC3 markedly suppressed the levels of IL-6,IL-8,monocyte chemoatbactant protein-1(MCP-1),p-p65,and p-IκBαin endothelial cells stimulated with IL-1β(P<0.01).Conversely,knockdown of NLRC3 resulted in elevated levels of IL-6,IL-8,MCP-1,p-p65 and p-IκBαin endothelial cells(P<0.01).Coronary artery disease patients had significantly lower plasma NLRC3 lev

关 键 词：NLR家族含CARD域蛋白3 内皮细胞 炎症 冠心病 NF-ΚB信号通路 

分 类 号：R541.4[医药卫生—心血管疾病] R329.21[医药卫生—内科学] R363[医药卫生—临床医学]