Fibrates down-regulate IL-1-stimulated C-reactive protein gene expression in hepatocytes by reducing nuclear p50-NFkappa B-C/EBP-beta complex formation

Blood. 2003 Jan 15;101(2):545-51. doi: 10.1182/blood-2002-06-1762. Epub 2002 Aug 29.

Abstract

C-reactive protein (CRP) is a major acute-phase protein in humans. Elevated plasma CRP levels are a risk factor for cardiovascular disease. CRP is predominantly expressed in hepatocytes and is induced by interleukin-1 (IL-1) and IL-6 under inflammatory situations, such as the acute phase. Fibrates are hypolipidemic drugs that act through the nuclear receptor peroxisome proliferator-activated receptor-alpha (PPAR-alpha). Fibrates have been shown to reduce elevated CRP levels in humans, but the molecular mechanism is unknown. In this study, we demonstrate that different PPAR-alpha activators suppress IL-1-induced, but not IL-6-induced, expression of CRP in primary human hepatocytes and HuH7 hepatoma cells. Induction of CRP expression by IL-1 occurs at the transcriptional level. Site-directed mutagenesis experiments show that IL-1 induces CRP expression through 2 overlapping response elements, the binding sites for CCAAT-box/enhancer-binding protein-beta (C/EBP-beta) and p50-nuclear factor-kappaB (p50-NFkappaB). Cotransfection of C/EBP-beta and p50-NFkappaB enhances CRP promoter activity, and coimmunoprecipitation experiments indicate that the increase in CRP promoter activity by IL-1 is related to the generation and nuclear accumulation of C/EBP-beta-p50-NFkappaB complexes. Interestingly, PPAR-alpha activators reduce the formation of nuclear C/EBP-beta-p50-NFkappaB complexes, and thereby CRP promoter activity, by 2 mechanisms. First, PPAR-alpha increases IkappaB-alpha expression and thus prevents p50-NFkappaB translocation to the nucleus. Second, fibrates decrease hepatic C/EBP-beta and p50-NFkappaB protein levels in mice in a PPAR-alpha-dependent way. Our findings identify C/EBP-beta and p50-NFkappaB as novel targets for PPAR-alpha and provide a molecular explanation for the reduction of plasma CRP levels by fibrates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bezafibrate / pharmacology*
  • C-Reactive Protein / drug effects
  • C-Reactive Protein / genetics*
  • CCAAT-Enhancer-Binding Protein-beta / genetics
  • CCAAT-Enhancer-Binding Protein-beta / metabolism*
  • Cell Nucleus / metabolism
  • Clofibric Acid / analogs & derivatives*
  • Clofibric Acid / pharmacology*
  • Down-Regulation / drug effects
  • Fibric Acids
  • Hepatocytes / drug effects
  • Hepatocytes / metabolism*
  • Humans
  • Hypolipidemic Agents / pharmacology*
  • Interleukin-1 / pharmacology*
  • Mice
  • Mice, Knockout
  • NF-kappa B / genetics
  • NF-kappa B / metabolism*
  • NF-kappa B p50 Subunit
  • Protein Binding / drug effects
  • Receptors, Cytoplasmic and Nuclear / genetics
  • Receptors, Cytoplasmic and Nuclear / metabolism
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Transcription, Genetic / drug effects
  • Transfection

Substances

  • CCAAT-Enhancer-Binding Protein-beta
  • Fibric Acids
  • Hypolipidemic Agents
  • Interleukin-1
  • NF-kappa B
  • NF-kappa B p50 Subunit
  • Receptors, Cytoplasmic and Nuclear
  • Transcription Factors
  • Clofibric Acid
  • C-Reactive Protein
  • ciprofibrate
  • Bezafibrate