摘要
目的:通过分析活化Toll样受体4(TLR4)对人脐静脉内皮细胞分泌炎性细胞因子的影响,探讨静脉内皮细胞在炎症反应中的作用。方法:逆转录聚合酶链式反应检测TLR4和细胞因子信使核糖核酸(mRNA)在人脐静脉内皮细胞中的表达,流式细胞分析检测TLR4及其辅助受体CD14的蛋白表达水平,酶联免疫吸附试验检测细胞培养上清中细胞因子的表达水平。结果:TLR4配体脂多糖诱导了人脐静脉内皮细胞中TLR4的基因和蛋白表达上调以及CD14的蛋白表达上调。活化TLR4显著上调人脐静脉内皮细胞中炎性细胞因子白细胞介素(IL)-1β和IL-8的基因表达以及IL-6和肿瘤坏死因子(TNF)-α的蛋白表达,并呈剂量依赖关系。活化TLR4诱导的细胞因子上调依赖NF-κB信号传导途径,信号传导阻滞剂能抑制细胞因子的表达上调。而且初次活化后TLR4的再次活化仍然诱导高水平的炎性细胞因子。结论:活化TLR4诱导静脉内皮细胞分泌炎性细胞因子,介导非耐受性炎症反应,提示静脉内皮细胞是重要的炎症效应细胞。
Objective ：To explore the roles of vein endothelial cells in the inflammatory response by analyzing the effect of the TLR4 activation on the secretion of pro-inflammation cytokines in human umbilical vein endothelial ceils （HUVECs）. Methods ： RT-PCR was used to detect the mRNA expression of TLR4 and cytokines in HUVECs. FACS was applied to detect the protein expression on the cell surface. The concentrations of pro-inflammation cytokines secreted in the supernatant of cultured cells were measured by ELISA. Results ：TLR4 ligand LPS induced the up-regulation of TLR4 gene and protein expression, and CD14 protein expression in HUVECs. TLR4 activation by ligand LPS significantly up-regulated the gene expression of pro-inflammation cytokines IL-1β and IL-8 ,and the protein expression of IL-6 and TNF-α in a dose-dependent manner. The up-regulation of cytokines induced by LPS depended on NF-κB signal pathway. NF-κB inhibitor repressed the secretion of cytokines. Moreover, the re-activation of TLR4 still induced high levels of pro-inflammation cytokines. Conclusion ： The activated TLR4 induced the secretion of pro-inflammation cytokines in vein endothelial cells and triggered a non-tolerant inflammatory response. These results suggested that vein endothelial cells were the important inflammatory effect cells.
出处
《中国循环杂志》
CSCD
北大核心
2009年第2期127-130,共4页
Chinese Circulation Journal
