Effect of acute and chronic inflammatory stimuli on expression of protease-activated receptors 1 and 2 in alveolar macrophages

J Allergy Clin Immunol. 2003 Feb;111(2):367-73. doi: 10.1067/mai.2003.6.

Abstract

Background: Protease-activated receptors 1 and 2 (PAR-1 and PAR-2) are 7-transmembrane G protein-coupled receptors activated by serine proteases in many cell types, including monocytes-macrophages, leading to the production of pro-inflammatory mediators, cytokines, and growth factors.

Objective: We determined the influence of chronic smoking and asthma on the expression of PAR-1 and PAR-2 receptors on alveolar macrophages (AMs).

Methods: We used RT-PCR and immunocytochemistry with confocal microscopy to determine mRNA and protein expression of PAR-1 and PAR-2 in AMs obtained from healthy smokers, asthmatic patients, and healthy subjects. In addition, we examined the effect of IL-1beta and LPS.

Results: PAR1 mRNA was decreased, whereas PAR2 mRNA was increased in 24-hour cultured AMs from smokers when compared with values in AMs from healthy subjects. Paradoxically, there was a higher degree of PAR-1 protein staining in AMs from smokers, whereas PAR-2 staining was similar in smokers and healthy subjects. PAR-1 and PAR-2 mRNA and protein expression were similar in asthmatic patients and control subjects. IL-1beta and LPS had no effect on PAR1 and PAR2 gene expression by AMs.

Conclusions: There is a dissociation between gene and protein expression of PAR-1 and PAR-2. PAR-1 protein overexpression in AMs from smokers might be important in the pathophysiology of chronic airways disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Asthma / genetics
  • Asthma / metabolism
  • Base Sequence
  • Case-Control Studies
  • Cells, Cultured
  • Female
  • Gene Expression / drug effects
  • Humans
  • Immunohistochemistry
  • In Vitro Techniques
  • Inflammation / genetics
  • Inflammation / metabolism*
  • Interleukin-1 / pharmacology
  • Lipopolysaccharides / pharmacology
  • Macrophages, Alveolar / drug effects
  • Macrophages, Alveolar / metabolism*
  • Male
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Receptor, PAR-1
  • Receptor, PAR-2
  • Receptors, Thrombin / genetics*
  • Receptors, Thrombin / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Smoking / genetics
  • Smoking / metabolism

Substances

  • Interleukin-1
  • Lipopolysaccharides
  • RNA, Messenger
  • Receptor, PAR-1
  • Receptor, PAR-2
  • Receptors, Thrombin