Title |
Human lung myofibroblast TGFβ1-dependent Smad2/3 signalling is Ca2+-dependent and regulated by KCa3.1 K+ channels
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Published in |
Fibrogenesis & Tissue Repair, March 2015
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DOI | 10.1186/s13069-015-0022-0 |
Pubmed ID | |
Authors |
Katy M Roach, Carol Feghali-Bostwick, Heike Wulff, Yassine Amrani, Peter Bradding |
Abstract |
Idiopathic pulmonary fibrosis (IPF) is a common and invariably lethal interstitial lung disease with poorly effective therapy. Blockade of the K(+) channel KCa3.1 reduces constitutive α-SMA and Smad2/3 nuclear translocation in IPF-derived human lung myofibroblasts (HLMFs), and inhibits several transforming growth factor beta 1 (TGFβ1)-dependent cell processes. We hypothesized that KCa3.1-dependent cell processes also regulate the TGFβ1-dependent Smad2/3 signalling pathway in HLMFs. HLMFs obtained from non-fibrotic controls (NFC) and IPF lungs were grown in vitro and examined for αSMA expression by immunofluorescence, RT-PCR, and flow cytometry. Two specific and distinct KCa3.1 blockers (TRAM-34 200 nM and ICA-17043 [Senicapoc] 100 nM) were used to determine their effects on TGFβ1-dependent signalling. Expression of phosphorylated and total Smad2/3 following TGFβ1 stimulation was determined by Western blot and Smad2/3 nuclear translocation by immunofluorescence. KCa3.1 block attenuated TGFβ1-dependent Smad2/3 phosphorylation and nuclear translocation, and this was mimicked by lowering the extracellular Ca(2+) concentration. KCa3.1 block also inhibited Smad2/3-dependent gene transcription (αSMA, collagen type I), inhibited KCa3.1 mRNA expression, and attenuated TGFβ1-dependent αSMA protein expression. KCa3.1 activity regulates TGFβ1-dependent effects in NFC- and IPF-derived primary HLMFs through the regulation of the TGFβ1/Smad signalling pathway, with promotion of downstream gene transcription and protein expression. KCa3.1 blockers may offer a novel approach to treating IPF. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Unknown | 30 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Master | 4 | 13% |
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Student > Bachelor | 3 | 10% |
Student > Ph. D. Student | 2 | 7% |
Other | 2 | 7% |
Unknown | 8 | 27% |
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Agricultural and Biological Sciences | 4 | 13% |
Chemistry | 1 | 3% |
Other | 0 | 0% |
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