摘要
目的研究THP-1细胞转化的巨噬细胞中Notch信号通路成分对氧化性低密度脂蛋白(oxidized low-density lipoprotein,ox-LDL)作用下的应答变化,以及Toll样受体2小分子干扰RNA(toll like receptor 2 small interfering RNA,TLR2siRNA)对该应答的作用,为了解Notch信号在炎症反应及参与动脉硬化机制提供实验依据。方法将贴壁的THP-1细胞分为3组:(1)诱导组:采用50 mg/L ox-LDL刺激转化贴壁的THP-1源巨噬细胞;(2)转染组:采用50 mg/L ox-LDL刺激转化贴壁的THP-1源巨噬细胞+TLR2siRNA转染;(3)对照组:转化并贴壁的THP-1源巨噬细胞。采用实时定量聚合酶链式反应(quantitative real time polymerase chain reaction,qPCR)检测Notch信号通路受体、配体的表达,单核细胞趋化蛋白1(monocyte chemoattractant protein 1,MCP-1)mRNA,血管细胞黏附分子1(vascular cell adhesive molecule 1,VCAM-1)mRNA的表达。将TLR2siRNA基因沉默后用Lipo2000转染THP-1源巨噬细胞,在ox-LDL作用下,qPCR分别检测TLR2siRNA转染的THP-1源巨噬细胞与未转染的THP-1源巨噬细胞的Notch信号成分及VCAM-1、MCP-1 mRNA表达。结果与对照组比较,加入ox-LDL后,THP-1源巨噬细胞表达Notch 1、DLL 4和Jagged 1表达明显升高(P<0.05),VCAM-1、MCP-1 mRNA的表达也明显升高(P<0.05);而接受TLR2siRNA转染后,可部分逆转上述ox-LDL对THP-1源巨噬细胞的致损活化效应。结论TLR2siRNA可抑制ox-LDL致损THP-1源巨噬细胞Notch信号通路各成分及VCAM-1与MCP-1 mRNA的表达上升。
Objective To study the response of Notch signaling pathway in the THP-1 transformed macrophages to oxidized low-density lipoprotein(ox-LDL),and to explore the effect of the toll-like receptor 2 small interfering RNA(TLR2 siRNA)on this response,and to provides experimental basis for understanding the role of Notch signal in inflammatory response and the mechanism involved in atherosclerosis.Methods The adherent THP-1 cells were divided into three groups,(1)Induced group:THP1-derived macrophages were stimulated by ox-LDL at the concentration of 50 mg/L;(2)Transfection group:THP1-derived macrophages were stimulated by ox-LDL at the concentration of 50 mg/L and were transferred by TLR2 siRNA;(3)Control group:transformed and adherent THP-1 source macrophages.The expression of Notch signaling pathway receptors and ligands,monocyte chemotactic protein 1(MCP-1)and vascular cell adhesion molecule 1(VCAM-1)mRNA were detected by quantitative real time polymerase chain reaction(qPCR).THP-1 macrophages were transfected with Lipo2000 after TLR2 siRNA gene silencing.Under the action of ox-LDL,the Notch signal components and the expression of VCAM-1 mRNA and MCP-1 mRNA in THP-1 derived macrophages and untransfected THP-1 derived macrophages were detected by qPCR.Results The expression of Notch 1,DLL 4,Jagged 1,VCAM-1 and MCP-1 mRNA in induced group was significantly higher than those of the control group(P<0.05).After TLR2 siRNA transfection,the damaged activation effect of ox-LDL on THP-1 macrophages could be partially reversed.Conclusion TLR2 siRNA can inhibit the increase of Notch signaling pathway and the expression of VCAM-1 and MCP-1 mRNA in THP-1 derived macrophages induced by ox-LDL.
作者
付文波
丁世芳
陈志楠
李志刚
卢青
FU Wenbo;DING Shifang;CHEN Zhinan;LI Zhigang;LU Qing(Department of Cardiology,General Hospital of Central Theater Command,Wuhan Hubei 430070,China)
出处
《华南国防医学杂志》
CAS
2020年第7期451-454,459,共5页
Military Medical Journal of South China
