异常血流动力对TLR4/NF—κB信号传导通路及其下游炎症因子的影响(1)
[摘 要] 目的 研究异常血流应力或压力单独作用对人脐静脉内皮细胞(Human umbilical vein endothelial cells，HUVECs)Toll样受体4(Toll-like receptor 4，TLR4)/NF-κB信号传导通路及下游炎症因子：血凝素样氧化低密度脂蛋白受体-1(lectin-like oxidized low-density lipoprotein receptor-1，LOX-1)、肿瘤坏死因子-α(TNF-α)、细胞间粘附分子-1(ICAM-1)及血管细胞粘附分子-1(VCAM-1)等的影响，探讨异常血流动力导致动脉粥样硬化的机制。方法 将生长良好的HUVECs以1×105个/ml密度接种于细胞综合应力刺激实验系统(型号BIO-CCS13)中进行干预。将HUVECs按所受的应力不同分成应力组、压力组和正常组。在各组应力作用下培养一天后收集细胞备用，用qPCR方法测定TLR4、髓样分化因子88(myeloid differentiation factor 88，MyD88)、肿瘤坏死因子受体相关因子-6(tumor necrosis factor receptor-associated factor-6，TRAF-6)、血凝素样氧化低密度酯蛋白受体-1(LOX-1)、核因子-κB(NF-κB)、TNF-α、ICAM-1及VCAM-1因子的基因表达，用蛋白质印迹方法测定TLR4、NF-κB、LOX-1、TNF-α、ICAM-1及VCAM-1因子的蛋白表达。结果 与正常组比较，应力组和压力组TLR4、MyD88、TRAF-6、LOX-1、NF-κB、TNF-α、ICAM-1及VCAM-1mRNA表达水平显著升高(P<0.01)，TLR4、LOX-1、NF-κB、TNF-α、ICAM-1及VCAM-1蛋白表达显著升高，差异有统计学意义(P<0.01)。结论 异常血流动力导致动脉粥样硬化的机制可能与激活TLR4/NF-κB信号传导通路和增强下游炎症因子：LOX-1、TNF-α、ICAM-1及VCAM-1等的表达有关。
[关键词] Toll样受体4；壁面压力；张应力；壁面切应力；动脉粥样硬化
中图分类号：R331 文献标识码：A 文章编号：1009-816X(2016)06-0415-04
[Abstract] Objective To study the effect of abnormal blood flow stress or pressure solely on expressions of Toll-like receptor 4 (TLR4)/NF-κB signaling pathway and downstream inflammatory factors in human umbilical vein endothelial cells (HUVECs)， namely lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1)， tumor necrosis factor-α (TNF-α)， intercellular cell adhesion molecule-1 (ICAM-1)， vascular cell adhesion molecule-1 (VCAM-1)， as well as its possible mechanisms leading to atherosclerosis. Methods The HUVECs were divided into stress group， pressure group and normal group according to the different stresses. The 4th， 5th generation of HUVECs were inoculated in the cell synthetic stress stimulation test system (model bio-ccs13) at 1×105 cells/ml. The cells were collected respectively 24 hours after the stress of each group. The gene expressions of TLR4， myeloid differentiation factor 88 (MyD88)， tumor necrosis factor receptor-associated factor-6 (TRAF-6)， LOX-1， NF-κB， TNF-α， VCAM-1 and ICAM-1 were detected by qPCR， and the protein expressions of TLR4， NF-κB， LOX-1， TNF-α， ICAM-1 and VCAM-1 by Western blot. Results Compared with the normal group， mRNA expressions of TLR4， MyD88， TRAF-6， LOX-1， NF-κB， TNF-α， VCAM-1 and ICAM-1 increased significantly (P<0.01) in the stress group and pressure group； protein expressions of TLR4， NF-κB， LOX-1， TNF-α， VCAM-1 and ICAM-1 also increased significantly (P<0.01). The difference was statistically significant. Conclusions The mechanism of abnormal blood flow stress or pressure leading to atherosclerosis might be associated with the activation of TLR4/NF-κB signal transduction pathway and the enhancement of its downstream inflammatory factors， such as the expressions of LOX-1， TNF-α， VCAM-1 and ICAM-1., http://www.100md.com(朱晔斌 朱晔君 李淑珍)
[关键词] Toll样受体4；壁面压力；张应力；壁面切应力；动脉粥样硬化
中图分类号：R331 文献标识码：A 文章编号：1009-816X(2016)06-0415-04
[Abstract] Objective To study the effect of abnormal blood flow stress or pressure solely on expressions of Toll-like receptor 4 (TLR4)/NF-κB signaling pathway and downstream inflammatory factors in human umbilical vein endothelial cells (HUVECs)， namely lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1)， tumor necrosis factor-α (TNF-α)， intercellular cell adhesion molecule-1 (ICAM-1)， vascular cell adhesion molecule-1 (VCAM-1)， as well as its possible mechanisms leading to atherosclerosis. Methods The HUVECs were divided into stress group， pressure group and normal group according to the different stresses. The 4th， 5th generation of HUVECs were inoculated in the cell synthetic stress stimulation test system (model bio-ccs13) at 1×105 cells/ml. The cells were collected respectively 24 hours after the stress of each group. The gene expressions of TLR4， myeloid differentiation factor 88 (MyD88)， tumor necrosis factor receptor-associated factor-6 (TRAF-6)， LOX-1， NF-κB， TNF-α， VCAM-1 and ICAM-1 were detected by qPCR， and the protein expressions of TLR4， NF-κB， LOX-1， TNF-α， ICAM-1 and VCAM-1 by Western blot. Results Compared with the normal group， mRNA expressions of TLR4， MyD88， TRAF-6， LOX-1， NF-κB， TNF-α， VCAM-1 and ICAM-1 increased significantly (P<0.01) in the stress group and pressure group； protein expressions of TLR4， NF-κB， LOX-1， TNF-α， VCAM-1 and ICAM-1 also increased significantly (P<0.01). The difference was statistically significant. Conclusions The mechanism of abnormal blood flow stress or pressure leading to atherosclerosis might be associated with the activation of TLR4/NF-κB signal transduction pathway and the enhancement of its downstream inflammatory factors， such as the expressions of LOX-1， TNF-α， VCAM-1 and ICAM-1., http://www.100md.com(朱晔斌 朱晔君 李淑珍)