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Downregulation of protease activated receptor expression and cytokine production in P815 cells by RNA interference

Liya Qiao1, Huiyun Zhang, Shandong Wu

  • 1Allergy and Inflammation Research Institute, Shantou University Medical College, 22 Xin-ling Road, Shantou, Guangdong 515041, PR China. qlyabc@sina.com.cn

BMC Cell Biology
|September 8, 2009
PubMed
Abstract

Insights

Small interfering RNAs (siRNAs) targeting protease-activated receptors (PARs) effectively modulated PAR expression and cytokine release in mast cells. This confirms the role of PARs in allergic reactions.

Area of Science:

  • Immunology
  • Cell Biology
  • Molecular Biology

Background:

  • Protease-activated receptors (PARs) are G-protein coupled receptors involved in cellular signaling.
  • Mast cells express PARs and release cytokines upon activation.
  • Interleukin (IL)-12 and tryptase influence PAR expression and cytokine release in mast cells.

Purpose of the Study:

  • To investigate the role of PARs in mast cell activation and cytokine production.
  • To determine the efficacy of small interfering RNAs (siRNAs) in modulating PAR expression.
  • To assess the impact of PAR modulation on IL-4 and IL-6 release.

Main Methods:

  • Utilized RNA interference (RNAi) with specific siRNAs targeting PAR-1, PAR-2, and PAR-4 in P815 mast cells.
  • Transfected cells with siRNAs and analyzed PAR mRNA and protein expression at 24, 48, and 72 hours.
  • Stimulated cells with IL-12, trypsin, tryptase, or PAR-activating peptides (SLIGKV-NH2, SFLLR-NH2) to measure cytokine release.

Main Results:

  • siRNAs significantly downregulated PAR-1, PAR-2, and PAR-4 expression at both mRNA and protein levels.
  • Specific siRNAs reduced IL-12-induced changes in PAR expression.
  • siRNA targeting PAR-2 and PAR-1 significantly inhibited trypsin- and peptide-induced IL-4 release, and tryptase-induced IL-4 release.

Conclusions:

  • siRNAs effectively modulate PAR expression in mast cells.
  • PARs play a significant role in mast cell-mediated cytokine production, particularly IL-4 release.
  • These findings support the involvement of PARs in allergic inflammatory responses.

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