Zeiher A M, Fisslthaler B, Schray-Utz B, Busse R
Department of Internal Medicine IV, University of Frankfurt, Germany.
Circ Res. 1995 Jun;76(6):980-6. doi: 10.1161/01.res.76.6.980.
The recruitment of monocytes into the arterial wall is one of the earliest events in the pathogenesis of atherosclerosis. Since monocyte chemoattractant protein 1 (MCP-1) plays a key role in the subendothelial recruitment of monocytes, we tested whether nitric oxide (NO) modulates the expression of MCP-1 in cultured human endothelial cells. Inhibition of basal NO production by NG-nitro-L-arginine (L-NAG) upregulates endothelial MCP-1 mRNA expression (250 +/- 20%) and protein secretion. Exogenous addition of NO dose-dependently decreased MCP-1 mRNA expression and secretion. Changes in MCP-1 mRNA expression and protein secretion were paralleled by corresponding changes in chemotactic activity of cell-conditioned media for monocytes. An MCP-1 antibody reduced monocyte chemotactic activity by 85% and completely abolished the increased monocyte chemotactic activity induced by the inhibition of NO production. Elevation of endothelial cGMP levels had no significant effect on MCP-1 mRNA expression. Inhibition of basal endothelial NO production by L-NAG increased binding activity of a nuclear factor kappa B (NF-kappa B)-like transcriptional regulatory factor, whereas exogenous addition of NO decreased NF-kappa B-like binding activity during stimulation with tumor necrosis factor-alpha. Thus, NO modulates MCP-1 expression and monocyte chemotactic activity secreted by human umbilical vein endothelial cells (HUVECs) in culture. The activation of NF-kappa B-like transcriptional regulatory proteins by inhibition of NO suggests a molecular link between an oxidant-sensitive transcriptional regulatory mechanism and NO synthesis in HUVECs.
单核细胞募集到动脉壁是动脉粥样硬化发病机制中最早出现的事件之一。由于单核细胞趋化蛋白1(MCP-1)在单核细胞的内皮下募集过程中起关键作用,我们检测了一氧化氮(NO)是否调节培养的人内皮细胞中MCP-1的表达。用NG-硝基-L-精氨酸(L-NAG)抑制基础NO生成会上调内皮MCP-1 mRNA表达(250±20%)和蛋白分泌。外源性添加NO可剂量依赖性地降低MCP-1 mRNA表达和分泌。MCP-1 mRNA表达和蛋白分泌的变化与细胞条件培养基对单核细胞的趋化活性的相应变化平行。一种MCP-1抗体可使单核细胞趋化活性降低85%,并完全消除因抑制NO生成而诱导的单核细胞趋化活性增加。内皮细胞cGMP水平升高对MCP-1 mRNA表达无显著影响。L-NAG抑制基础内皮NO生成可增加一种核因子κB(NF-κB)样转录调节因子的结合活性,而外源性添加NO可在肿瘤坏死因子-α刺激期间降低NF-κB样结合活性。因此,NO调节培养的人脐静脉内皮细胞(HUVECs)分泌的MCP-1表达和单核细胞趋化活性。抑制NO导致NF-κB样转录调节蛋白激活,提示在HUVECs中氧化敏感转录调节机制与NO合成之间存在分子联系。