title: C-reactive protein induces NF-kappaB activation through intracellular calcium and ROS in human mesangial cells.
abstract: 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-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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Internal Medicine, Urology, Asan Institute for Life Sciences, College of Medicine, University of Ulsan, Seoul, South Korea.
authors.forename: Jai Won
authors.initials: JW
authors.lastname: Chang
authors.forename: Choung Soo
authors.initials: CS
authors.lastname: Kim
authors.forename: Soon Bae
authors.initials: SB
authors.lastname: Kim
authors.forename: Su Kil
authors.initials: SK
authors.lastname: Park
authors.forename: Jung Sik
authors.initials: JS
authors.lastname: Park
authors.forename: Sang Koo
authors.initials: SK
authors.lastname: Lee
mesh_terms: D014013:1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt / Q000494:pharmacology; D000111:Acetylcysteine / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D015152:Blotting, Northern; D002097:C-Reactive Protein / Q000494:pharmacology* / Q000502:physiology*; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology; D018932:Chemokine CCL2 / Q000235:genetics / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D024202:Electrophoretic Mobility Shift Assay; D005786:Gene Expression Regulation; D006801:Humans; D007249:Inflammation; D050527:Mesangial Cells / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D018613:Microscopy, Confocal; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism*; D011994:Recombinant Proteins / Q000494:pharmacology; D013997:Time Factors
chemical_list: D000975:Antioxidants; D002614:Chelating Agents; D018932:Chemokine CCL2; D016328:NF-kappa B; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D011994:Recombinant Proteins; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D014013:1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt; D004533:Egtazic Acid; D002097:C-Reactive Protein; D002118:Calcium; D000111:Acetylcysteine