Histamine induces ATP release from human subcutaneous fibroblasts via pannexin-...

• Autor(es): Pinheiro, Ana Rita ; Paramos-de-Carvalho, Diogo ; Certal, Mariana ; Costa, M. Adelina ; Costa, Cristina ; Magalhães-Cardoso, Maria Teresa ; Ferreirinha, Fátima ; Sévigny, Jean ; Correia-de-Sá, Paulo
• Data: 2013
• Identificador Persistente: http://hdl.handle.net/10400.22/3434
• Origem: Repositório Científico do Instituto Politécnico do Porto
• Assunto(s): ADP; ATP; ATP release; ATPases; Calcium; Human subcutaneous fibroblasts; Mechanotransduction; Nucleoside nucleotide metabolism; Pannexin; Purinergic receptor
• Tipo de Documento: Artigo
• Idioma: Inglês

Descrição

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 Ca2+ ([Ca2+]i) and ATP release from human subcutaneous fibroblasts via H1 receptor activation. Histamine-induced [Ca2+]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. [Ca2+]i accumulation caused by histamine was also reduced upon blocking pannexin-1 hemichannels with 10Panx, 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 [Ca2+]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 ADPsensitive 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 [Ca2+]i mobilization and cell growth through the cooperation of H1 and P2 (probably P2Y1) receptors.