Increased constitutive αSMA and Smad2/3 expression in idiopathic pulmonary fibrosis myofibroblasts is KCa3.1-dependent.
Increased constitutive αSMA and Smad2/3 expression in idiopathic pulmonary fibrosis myofibroblasts is KCa3.1-dependent.
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在特发性肺纤维化肌纤维细胞中增加的组成型αSMA和SMAD2/3表达是KCA3.1依赖性的。
DOI：
10.1186/s12931-014-0155-5
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发表时间：
2014-12-05
影响因子：
5.8
通讯作者：
Bradding P
中科院分区：
文献类型：
--
作者：
Roach KM;Wulff H;Feghali-Bostwick C;Amrani Y;Bradding P
特发性肺纤维化是一种常见且总是致命的疾病，治疗选择有限。钙激活的KCa3.1钾通道在促进人肺肌成纤维细胞（HLMF）的TGFβ1和bFGF依赖的促纤维化反应中起关键作用。我们假设KCa3.1通道依赖性细胞过程通过Smad 2/3信号通路调节HLMF αSMA表达。在这项研究中，我们比较了来自非纤维化健康对照肺（NFC）的HLMF与来自IPF肺的细胞的表型。采用免疫荧光（IF）、RT-PCR和流式细胞术检测αSMA的表达。通过RT-PCR、western blot和免疫荧光检测基础Smad 2/3信号。使用两种特异性和不同的KCa 3.1阻断剂（TRAM-34 200 nM和伊卡-17043 [Senicapoc] 100 nM）来确定其对HLMF分化和Smad 2/3信号传导通路的影响。IPF衍生的HLMF表现出α-平滑肌肌动蛋白（αSMA）和肌动蛋白应力纤维的组成性表达增加，表明肌成纤维细胞分化程度更高。这与Smad 2/3 mRNA和蛋白表达的增加以及Smad 2/3核定位的增加有关。通过清除细胞外Ca 2+或使用选择性KCa3.1阻断剂（TRAM-34、伊卡-17043）阻断KCa3.1离子通道，可抑制Smad 2/3核定位增加。这伴随着IPF衍生的HLMF向静止成纤维细胞表型的去分化，如αSMA表达减少和肌动蛋白应力纤维形成减少所证明的。两者合计，这些数据表明，Ca 2+和KCa3.1依赖的过程促进“组成型”Smad 2/3信号在IPF衍生的成纤维细胞，从而促进成纤维细胞分化成肌纤维细胞。重要的是，抑制KCa3.1通道逆转了这一过程。因此，靶向KCa3.1可能为治疗IPF提供一种新的有效方法，并且有可能将KCa3.1靶向治疗快速转化为临床治疗。
Idiopathic pulmonary fibrosis is a common and invariably fatal disease with limited therapeutic options. Ca2+-activated KCa3.1 potassium channels play a key role in promoting TGFβ1 and bFGF-dependent profibrotic responses in human lung myofibroblasts (HLMFs). We hypothesised that KCa3.1 channel-dependent cell processes regulate HLMF αSMA expression via Smad2/3 signalling pathways. In this study we have compared the phenotype of HLMFs derived from non-fibrotic healthy control lungs (NFC) with cells derived from IPF lungs. HLMFs grown in vitro were examined for αSMA expression by immunofluorescence (IF), RT-PCR and flow cytommetry. Basal Smad2/3 signalling was examined by RT-PCR, western blot and immunofluorescence. Two specific and distinct KCa3.1 blockers (TRAM-34 200 nM and ICA-17043 [Senicapoc] 100 nM) were used to determine their effects on HLMF differentiation and the Smad2/3 signalling pathways. IPF-derived HLMFs demonstrated increased constitutive expression of both α-smooth muscle actin (αSMA) and actin stress fibres, indicative of greater myofibroblast differentiation. This was associated with increased constitutive Smad2/3 mRNA and protein expression, and increased Smad2/3 nuclear localisation. The increased Smad2/3 nuclear localisation was inhibited by removing extracellular Ca2+ or blocking KCa3.1 ion channels with selective KCa3.1 blockers (TRAM-34, ICA-17043). This was accompanied by de-differentiation of IPF-derived HLMFs towards a quiescent fibroblast phenotype as demonstrated by reduced αSMA expression and reduced actin stress fibre formation. Taken together, these data suggest that Ca2+- and KCa3.1-dependent processes facilitate “constitutive” Smad2/3 signalling in IPF-derived fibroblasts, and thus promote fibroblast to myofibroblast differentiation. Importantly, inhibiting KCa3.1 channels reverses this process. Targeting KCa3.1 may therefore provide a novel and effective approach for the treatment of IPF and there is the potential for the rapid translation of KCa3.1-directed therapy to the clinic.
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DOI：
10.1016/j.biocel.2006.11.005
发表时间：
2007-01-01
影响因子：
4
作者：
McAnulty, Robin J.
通讯作者：
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2003-09-01
影响因子：
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10.1164/rccm.200602-163oc
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2006-10-01
影响因子：
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