摘要
为探讨Toll样受体 4 (Toll likereceptor 4 ,TLR4 )在内毒素 (LPS)对内皮细胞NF κB激活中的作用 ,以LPS刺激培养的ECV 30 4细胞为模型 ,运用RT PCR和蛋白质印迹技术检测了内皮细胞TLR4的表达及LPS对其表达的影响 .同时利用基因转染和抗体阻断方法进一步观察了TLR4在LPS对内皮细胞NF κB激活中的作用 .研究发现 ,LPS能明显上调内皮细胞TLR4的表达 ,呈一定的时间和剂量依赖性 .转染TLR4的功能突变体和运用抗TLR4单抗能明显抑制LPS对内皮细胞NF κB的激活 .提示TLR4介导了LPS对内皮细胞NF κB激活 ,可能在LPS对内皮细胞激活
In order to investigate the role of Toll-like recepter 4 (TLR4) in lipopolysaccharide (LPS)-induced NF-kappaB activation in human endothelial cells, LPS-stimulated ECV-304 cells were used as experimental model and the expression of TLR4 and effect of LPS on the expression were analysed with RT-PCR and Western blot assay. Moreover, the role of TLR4 in LPS-induced NF-kappaB activation in endothelial cells was explored with gene transfection of non-signaling mutant forms of TLR4 and anti-TLR4 monoclonal antibody. The results showed that LPS could upregulat the expression of TLR4 in time and dose-dependent manner and that transfection of non-signaling mutant forms of TLR4 and anti-TLR4 monoclonal antibody inhibited LPS-induced NF-kappaB activation in human endothelial cells obviously. These data indicated that TLR4 mediates LPS-induced NF-kappaB activation in human endothelial cells and it may play important role in endothelial cell activation and injury induced by LPS.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2002年第3期407-410,共4页
Progress In Biochemistry and Biophysics
基金
国家重点基础研究发展规划项目 (973 ) (G19990 5 42 0 3 )
全军医药卫生科研基金资助项目 (0 1Q10 5 ) .~~
