C-reactive protein induces NF-κB activation through intracellular calcium and ROS in human mesangial cells
C-reactive protein induces NF-κB activation through intracellular calcium and ROS in human mesangial cells
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DOI：
10.1159/000087940
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发表时间：
2005-01-01
影响因子：
--
通讯作者：
Lee, SK
中科院分区：
文献类型：
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作者：
Chang, JW;Kim, CS;Lee, SK
背景：已知C反应蛋白（CRP）在内皮细胞中具有直接的促炎作用。然而，人们对C反应蛋白对固有肾细胞的影响知之甚少。我们研究了CRP对人肾小球系膜细胞核因子-κ B（NF-κ B）活化和单核细胞趋化蛋白-1（MCP-1）基因表达的影响，并检测了细胞内钙离子和活性氧（ROS）是否参与CRP诱导的NF-κ B活化。研究方法：采用凝胶电泳迁移率改变法和北方印迹法分别检测NF-κ B结合活性和MCP-1 mRNA表达。使用钙敏感染料Fluo-3通过共聚焦显微镜监测细胞内钙，并使用2 '，7'-二氯荧光素二乙酸酯测定细胞内ROS产生。结果如下：CRP以剂量依赖性方式（12.5-100 μ g/ml）增加NF-κ B结合活性，在孵育后1 h内诱导，并在3 h左右达到峰值。CRP还通过激活NF-κ B B增加MCP-1 mRNA表达。CRP可诱导细胞内钙离子和ROS的产生。钙螯合剂BAPTA-AM和抗氧化剂如N-乙酰半胱氨酸和Tiron抑制CRP诱导的NF-κ B活化。结论：CRP通过NF-κ B B激活诱导MCP-1基因表达，在人肾小球系膜细胞中发挥促炎作用，NF-κ B激活至少部分通过细胞内钙离子和ROS介导。版权所有（C）2005 S. Karger AG，巴塞尔。
Background: C-reactive protein (CRP) is known to have a direct proinflammatory effect in endothelial cells. However, little is known about the effect of CRP in intrinsic renal cells. We investigated the effects of CRP on the nuclear factor-kappa B (NF-kappa B) activation and monocyte chemoattractant protein-1 (MCP-1) gene expression in human mesangial cells and also examined whether intracellular calcium and reactive oxygen species (ROS) were involved in the CRP-induced NF-kappa B activation. Methods: NF-kappa B binding activity and MCP-1 mRNA expression were measured by electrophoretic mobility shift assay and Northern blot analysis, respectively. Intracellular calcium was monitored by confocal microscopy using calcium sensitive dye, Fluo-3 and intracellular ROS production was determined, using 2',7'-dichlorofluorescin diacetate. Results: CRP increased NF-kappa B binding activity in a dose-dependent manner (12.5-100 mu g/ml), which was induced within 1 h after incubation and peaked around 3 h. CRP also increased the MCP-1 mRNA expression via activation of NF-kappa B. Both intracellular calcium and ROS was induced by CRP. Calcium chelator, BAPTA-AM and anti-oxidants such as N-acetylcysteine and tiron suppressed CRP-induced NF-kappa B activation. Conclusion: CRP exerted a proinflammatory effect in human mesangial cells by inducing MCP-1 gene expression via NF-kappa B activation, which was mediated, at least in part, through intracellular calcium and ROS. Copyright (C) 2005 S. Karger AG, Basel.