Macrophage conditioned medium induces the expression of C-reactive protein in human aortic endothelial cells: potential for paracrine/autocrine effects.
Macrophage conditioned medium induces the expression of C-reactive protein in human aortic endothelial cells: potential for paracrine/autocrine effects.
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DOI：
10.1016/s0002-9440(10)62345-0
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发表时间：
2005-04
期刊：
影响因子：
--
通讯作者：
S. Venugopal;S. Devaraj;I. Jialal
中科院分区：
文献类型：
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作者：
S. Venugopal;S. Devaraj;I. Jialal
C-反应蛋白（CRP）是表面健康人群心血管事件的危险标志物。有力的数据显示，除了肝脏，CRP在动脉粥样硬化病变、肾脏、神经元和肺泡巨噬细胞中产生。由于CRP的几种致动脉粥样硬化作用已被证明在内皮细胞中，我们研究了人主动脉内皮细胞（HAEC）的CRP生产。RT-PCR和原位杂交检测CRP mRNA的表达，Western blot检测细胞内蛋白的表达，ELISA检测分泌蛋白的表达。与细胞因子IL-1、IL-6和肿瘤坏死因子单独或联合孵育显示，IL-1和IL-6联合使用对HAEC产生CRP的作用最强（P < 0.05）。为了模拟体内情况，我们研究了血管平滑肌细胞（VSMC）和/或巨噬细胞条件培养基（MCM）是否可以增加HAEC产生CRP。尽管VSMC条件培养基没有影响，但与MCM孵育导致细胞内和分泌CRP的合成显著增加两倍（P < 0.05）。MCM的作用可通过抑制IL-1和IL-6而逆转。因此，动脉粥样硬化病变中细胞通过旁分泌/自分泌环刺激CRP的合成和分泌，可导致CRP的局部浓度远远超过血浆浓度，并可能导致促炎、促动脉粥样硬化效应。
C-reactive protein (CRP) is a risk marker for cardiovascular events in apparently healthy persons. Cogent data show that, aside from the liver, CRP is produced in atherosclerotic lesions, kidney, neurons, and alveolar macrophages. Because several proatherogenic effects of CRP have been documented in endothelial cells, we examined human aortic endothelial cells (HAEC) for CRP production. We detected the presence of CRP mRNA by RT-PCR and in situ hybridization, intracellular protein by Western blot and secreted protein by ELISA. Coincubation with the cytokines interleukin (IL)-1, IL-6, and tumor necrosis factor alone and in combination showed that the most potent agonist for CRP production from HAEC is the combination of IL-1 and IL-6 (P < 0.05). To mimic the in vivo situation, we examined whether vascular smooth muscle cell (VSMC) and/or macrophage conditioned media (MCM) could augment CRP production by HAEC. While VSMC-conditioned media had no effect, incubation with MCM resulted in a significant twofold increase in the synthesis of both intracellular and secreted CRP (P < 0.05). The effect of MCM could be reversed by inhibiting both IL-1 and IL-6. Thus, stimulated synthesis and secretion of CRP by cells in the atherosclerotic lesion by paracrine/autocrine loops could result in local concentrations of CRP far in excess of plasma concentrations and could contribute to proinflammatory, proatherogenic effects.