Chloride channel activity in human lung fibroblasts and myofibroblasts

Zhaohong Yin, Mitchell A. Watsky

Research output: Contribution to journalArticle

31 Scopus citations

Abstract

It is well established that transforming growth factor (TGF)-β stimulates human lung fibroblasts (HLF) to differentiate into myofibroblasts. We characterized lysophosphatidic acid (LPA)-activated Cl- channel current (ICl-LPA) in cultured human lung fibroblasts and myofibroblasts and investigated the influence of ICl-LPA on fibroblast-to-myofibroblast differentiation. We recorded ICl-LPA using the amphotericin perforated-patch technique. We activated I Cl-LPA using LPA or sphingosine-1-phosphate. We determined phenotype by Western blotting and immunohistochemistry using an anti-α-smooth muscle actin (SMA) antibody. RT-PCR was performed to determine which phospholipid growth factor receptors are present in HLF. We found that HLF cultured in TGF-β (myofibroblasts) had significantly elevated α-SMA levels and ICl-LPA current density compared with control fibroblasts. I Cl-LPA activation was blocked by DIDS, 5-nitro-2-(3- phenylpropylamino)benzoic acid (NPPB), and the LPA receptor-specific antagonist dioctyl-glycerol pyrophosphate (1 μM). DIDS and NPPB, in a dose-dependent manner, significantly reduced α-SMA levels in HLF stimulated with TGF-β. These results demonstrate the receptor-mediated activation of I Cl-LPA by LPA and sphingosine-1-phosphate in cultured human lung myofibroblasts, with only minimal ICl-LPA activity in fibroblasts. This Cl- channel activity appears to play a critical role in the differentiation of human lung fibroblasts to myofibroblasts.

Original languageEnglish (US)
Pages (from-to)L1110-L1116
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume288
Issue number6 32-6
DOIs
Publication statusPublished - Jun 1 2005
Externally publishedYes

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Keywords

  • Lysophosphatidic acid
  • Sphingosine-1-phosphate
  • Transforming growth factor-β
  • α-smooth muscle actin

ASJC Scopus subject areas

  • Physiology
  • Pulmonary and Respiratory Medicine
  • Physiology (medical)
  • Cell Biology

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