Histamine induces ATP release from human subcutaneous fibroblasts, via pannexin-1 hemichannels, leading to Ca2+ mobilization and cell proliferation

J Biol Chem. 2013 Sep 20;288(38):27571-27583. doi: 10.1074/jbc.M113.460865. Epub 2013 Aug 5.

Abstract

Changes in the regulation of connective tissue ATP-mediated mechano-transduction and remodeling may be an important link to the pathogenesis of chronic pain. It has been demonstrated that mast cell-derived histamine plays an important role in painful fibrotic diseases. Here we analyzed the involvement of ATP in the response of human subcutaneous fibroblasts to histamine. Acute histamine application caused a rise in intracellular Ca(2+) ([Ca(2+)]i) and ATP release from human subcutaneous fibroblasts via H1 receptor activation. Histamine-induced [Ca(2+)]i rise was partially attenuated by apyrase, an enzyme that inactivates extracellular ATP, and by blocking P2 purinoceptors with pyridoxal phosphate-6-azo(benzene-2,4-disulfonic acid) tetrasodium salt and reactive blue 2. [Ca(2+)]i accumulation caused by histamine was also reduced upon blocking pannexin-1 hemichannels with (10)Panx, probenecid, or carbenoxolone but not when connexin hemichannels were inhibited with mefloquine or 2-octanol. Brefeldin A, an inhibitor of vesicular exocytosis, also did not block histamine-induced [Ca(2+)]i mobilization. Prolonged exposure of human subcutaneous fibroblast cultures to histamine favored cell growth and type I collagen synthesis via the activation of H1 receptor. This effect was mimicked by ATP and its metabolite, ADP, whereas the selective P2Y1 receptor antagonist, MRS2179, partially attenuated histamine-induced cell growth and type I collagen production. Expression of pannexin-1 and ADP-sensitive P2Y1 receptor on human subcutaneous fibroblasts was confirmed by immunofluorescence confocal microscopy and Western blot analysis. In conclusion, histamine induces ATP release from human subcutaneous fibroblasts, via pannexin-1 hemichannels, leading to [Ca(2+)]i mobilization and cell growth through the cooperation of H1 and P2 (probably P2Y1) receptors.

Keywords: ADP; ATP; ATP Release; ATPases; Calcium; Human Subcutaneous Fibroblasts; Mechanotransduction; Nucleoside Nucleotide Metabolism; Pannexin; Purinergic Receptor.

Publication types

  • Clinical Trial
  • Multicenter Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Diphosphate / analogs & derivatives
  • Adenosine Diphosphate / metabolism
  • Adenosine Diphosphate / pharmacology
  • Adenosine Triphosphate / metabolism*
  • Anti-Bacterial Agents / pharmacology
  • Antimalarials / pharmacology
  • Brefeldin A / pharmacology
  • Calcium / metabolism*
  • Cell Proliferation / drug effects*
  • Cells, Cultured
  • Collagen Type I / biosynthesis
  • Connexins / antagonists & inhibitors
  • Connexins / metabolism*
  • Exocytosis / drug effects
  • Exocytosis / physiology
  • Female
  • Fibroblasts / cytology
  • Fibroblasts / metabolism*
  • Gene Expression Regulation / drug effects
  • Gene Expression Regulation / physiology
  • Histamine / metabolism
  • Histamine / pharmacology*
  • Histamine Agonists / metabolism
  • Histamine Agonists / pharmacology*
  • Humans
  • Male
  • Mast Cells / cytology
  • Mast Cells / metabolism
  • Mefloquine / pharmacology
  • Middle Aged
  • Nerve Tissue Proteins / antagonists & inhibitors
  • Nerve Tissue Proteins / metabolism*
  • Octanols / pharmacology
  • Purinergic P2Y Receptor Antagonists / pharmacology
  • Receptors, Histamine H1 / metabolism
  • Receptors, Purinergic P2Y1 / metabolism

Substances

  • Anti-Bacterial Agents
  • Antimalarials
  • Collagen Type I
  • Connexins
  • Histamine Agonists
  • N(6)-methyl-2'-deoxyadenosine 3',5'-diphosphate
  • Nerve Tissue Proteins
  • Octanols
  • PANX1 protein, human
  • Purinergic P2Y Receptor Antagonists
  • Receptors, Histamine H1
  • Receptors, Purinergic P2Y1
  • Brefeldin A
  • Adenosine Diphosphate
  • 2-octanol
  • Histamine
  • Adenosine Triphosphate
  • Calcium
  • Mefloquine