Noncanonical STAT3 activation regulates excess TGF-β1 and collagen I expression in muscle of stricturing Crohn's disease.
Noncanonical STAT3 activation regulates excess TGF-β1 and collagen I expression in muscle of stricturing Crohn's disease.
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DOI：
10.4049/jimmunol.1401779
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发表时间：
2015-04-01
期刊：
影响因子：
--
通讯作者：
Kuemmerle JF
中科院分区：
文献类型：
--
作者：
Li C;Iness A;Yoon J;Grider JR;Murthy KS;Kellum JM;Kuemmerle JF
肠间充质肌细胞中TGF-β1和TGF-β1依赖性胶原I的产生增加导致蒙特利尔B2纤维狭窄性克罗恩病患者的纤维化许多细胞因子，包括IL-6，由活化的间充质细胞本身产生并激活STAT 3。本研究的目的是确定STAT-3激活可能导致肠纤维化的机制。通过ELISA测量细胞因子水平。通过免疫印迹法测定STAT 3和SOCS 3蛋白水平，通过ChIP法测定STAT 3-TGFB 1 DNA结合活性，通过荧光素酶报告基因测定法测定TGFB 1转录活性。采用qRT-PCR检测TGF-β1、胶原1 α1和CTGF的表达。使用STAT 3抑制剂Stattic和通过转染STAT 3突变体来确定STAT 3活化的作用。与其他克罗恩病表型、溃疡性结肠炎和非克罗恩病患者相比，来自同一患者的狭窄和正常肠的B2表型患者的肌细胞中细胞因子的自分泌产生增加。出现了一种独特的STAT 3磷酸化模式：狭窄中的高STAT 3（S727）和低STAT 3（Y 705），而未受影响的肠中则相反。TGFB 1的转录活性受磷酸化STAT 3（S727）的调节，并被Stattic或dnSTAT 3（S727 A）降低。TGF-β1、COL 1A 1和CTGF的表达被Stattic或dnSTAT 3（S727 A）抑制。用IL-6或表达组成型活性STAT 3（S727 E）表型复制的来自狭窄肠的肌细胞处理正常肌细胞。中和自分泌IL-6逆转狭窄肠肌中STAT 3磷酸化和TGF-β1。在TNBS诱导的结肠炎小鼠中证实了Stattic改善纤维化发展的能力。我们在患有蒙特利尔B2克罗恩病的患者中观察到独特的p-STAT 3（S727）应答，特别是在对IL-6的应答中，导致回肠狭窄中TGF-β1、胶原和CTGF产生增加。
Increased TGF-β1 and TGF-β1-dependent Collagen I production in intestinal mesenchymal muscle cells result in fibrosis in patients with Montreal B2 fibrostenotic Crohn's disease. Numerous cytokines, including IL-6, are produced by activated mesenchymal cells themselves and activate STAT3. The aim of the present study was to determine the mechanisms by which STAT-3-activation might result in intestinal fibrosis. Cytokine levels were measured by ELISA. STAT3 and SOCS3 protein levels were measured by immunoblot, STAT3-TGFB1 DNA-binding activity by ChIP, and TGFB1 transcriptional activity by luciferase reporter assay. TGF-β1, Collagen1α1 and CTGF expression was measured by qRT-PCR. The role of STAT3 activation was determined using STAT3 inhibitor, Stattic, and by transfection of STAT3 mutants. Autocrine production of cytokines was increased in muscle cells of B2 phenotype patients from strictures and normal intestine in the same patient and compared to other Crohn's phenotypes, ulcerative colitis and non-Crohn's patients. A unique pattern of STAT3 phosphorylation emerged: high STAT3(S727) and low STAT3(Y705) in strictures and the opposite in unaffected intestine. TGFB1 transcriptional activity was regulated by phospho-STAT3(S727) and was decreased by Stattic or dnSTAT3(S727A). TGF-β1, COL1A1, and CTGF expression was inhibited by Stattic or dnSTAT3(S727A). Treatment of normal muscle cells with IL-6 or expression of constitutively-active STAT3(S727E) phenocopied muscle cells from strictured intestine. Neutralization of autocrine IL-6 reversed STAT3 phosphorylation and TGF-β1 in strictured intestinal muscle. The ability of Stattic to improve development of fibrosis was confirmed in mice with TNBS-induced colitis. We observed a unique p-STAT3(S727) response in patients with Montreal B2 Crohn's disease particularly in response to IL-6 leading to increased TGF-β1, collagen and CTGF production in ileal strictures.
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影响因子：
30.8
作者：
通讯作者：
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影响因子：
29.4
作者：
GRAHAM, MF;BRYSON, GR;DIEGELMANN, RF
通讯作者：
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DOI：
10.1152/ajpgi.00310.2002
发表时间：
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影响因子：
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作者：
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通讯作者：
Kuemmerle, JF
影响因子：
29.4
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影响因子：
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作者：
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