C-reactive protein induces NF-kappaB activation through intracellular calcium and ROS in human mesangial cells

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-kappaB (NF-kappaB) 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-kappaB activation.

NF-kappaB 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.

CRP increased NF-kappaB binding activity in a dose-dependent manner (12.5-100 microg/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-kappaB. 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-kappaB activation.

CRP exerted a proinflammatory effect in human mesangial cells by inducing MCP-1 gene expression via NF-kappaB activation, which was mediated, at least in part, through intracellular calcium and ROS.