C-reactive protein triggers inflammatory responses partly via TLR4/IRF3/NF-κB signaling pathway in rat vascular smooth muscle cells

Life Sci. 2010 Sep 11;87(11-12):367-74. doi: 10.1016/j.lfs.2010.07.012. Epub 2010 Aug 4.

Abstract

Aims: C-reactive protein (CRP) plays an important role in the inflammatory process of atherosclerosis. Toll-like receptor 4 (TLR4) participates in atherogenesis by mediating the inflammatory responses. The aim of this experiment was to investigate the pro-inflammatory effects and mechanisms of CRP in rat vascular smooth muscle cells (VSMCs), especially focusing on the effects of CRP on IL-6 and peroxisome proliferator-activated receptor γ (PPARγ), and TLR4-dependent signal pathway.

Main methods: rat VSMCs were cultured, and CRP was used as a stimulant for IL-6 and peroxisome proliferator-activated receptor γ (PPARγ). IL-6 level in the culture supernatant was measured by ELISA, and mRNA and protein expressions were assayed by quantitative real-time PCR and western blot, respectively. RNA interference was used to assess the roles of TLR4 and interferon regulatory factor 3 (IRF3) in the pro-inflammatory signal pathway of CRP.

Key findings: CRP stimulated IL-6 secretion, and inhibited mRNA and protein expression of PPARγ in VSMCs in a concentration-dependent manner. Additionally, CRP induced TLR4 expression, promoted nuclear translocation of NF-κB (p65), and augmented IκBα phosphorylation in VSMCs. Taken together, CRP induces the inflammatory responses through increasing IL-6 generation and reducing PPARγ expression in VSMCs, which is mediated by TLR4/IRF3/NF-κB signal pathway.

Significance: CRP is able to stimulate IL-6 production and to inhibit PPARγ expression in VSMCs via MyD88-independent TLR4 signaling pathway (TLR4/IRF3/NF-κB). These provide the novel evidence for the pro-inflammatory action of CRP involved in atherogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • C-Reactive Protein / pharmacology*
  • C-Reactive Protein / physiology
  • Cells, Cultured
  • Gene Expression Regulation / drug effects
  • Gene Expression Regulation / genetics
  • Humans
  • I-kappa B Proteins / metabolism
  • Inflammation / chemically induced
  • Inflammation / metabolism
  • Interferon Regulatory Factor-3 / genetics
  • Interferon Regulatory Factor-3 / metabolism*
  • Interleukin-6 / biosynthesis
  • Interleukin-6 / metabolism
  • Lipopolysaccharides / pharmacology
  • Male
  • Muscle, Smooth, Vascular / cytology*
  • Myocytes, Smooth Muscle / drug effects*
  • Myocytes, Smooth Muscle / metabolism
  • NF-KappaB Inhibitor alpha
  • NF-kappa B / antagonists & inhibitors
  • NF-kappa B / metabolism*
  • PPAR gamma / genetics
  • PPAR gamma / metabolism
  • Phosphorylation / drug effects
  • Polymyxin B / pharmacology
  • Proline / analogs & derivatives
  • Proline / pharmacology
  • RNA, Small Interfering / genetics
  • Rats
  • Rats, Sprague-Dawley
  • Signal Transduction / drug effects*
  • Signal Transduction / physiology
  • Thiocarbamates / pharmacology
  • Toll-Like Receptor 4 / genetics
  • Toll-Like Receptor 4 / metabolism*

Substances

  • I-kappa B Proteins
  • Interferon Regulatory Factor-3
  • Interleukin-6
  • Lipopolysaccharides
  • NF-kappa B
  • NFKBIA protein, human
  • Nfkbia protein, rat
  • PPAR gamma
  • RNA, Small Interfering
  • Thiocarbamates
  • Tlr4 protein, rat
  • Toll-Like Receptor 4
  • prolinedithiocarbamate
  • NF-KappaB Inhibitor alpha
  • C-Reactive Protein
  • Proline
  • Polymyxin B