Eickelberg O, Pansky A, Koehler E, Bihl M, Tamm M, Hildebrand P, Perruchoud A P, Kashgarian M, Roth M
Department of Pathology, Yale University School of Medicine, New Haven, Connecticut 06520-8023, USA.
FASEB J. 2001 Mar;15(3):797-806. doi: 10.1096/fj.00-0233com.
Lung fibrosis is a fatal condition of excess extracellular matrix (ECM) deposition associated with increased transforming growth factor beta (TGF-beta) activity. Although much is known about its pathological features, our understanding of the signal transduction pathways resulting in increased ECM and collagen deposition in response to TGF-beta is still incompletely defined. We have previously reported that a JunD homodimer of the transcription factor AP-1 is specifically activated by TGF-beta in lung fibroblasts. Here we demonstrate that JunD is also specifically required for TGF-beta-induced effects. Antisense against JunD, but not c-fos or c-jun, significantly inhibited collagen deposition in response to TGF-beta in primary human lung fibroblasts. We then investigated the ability of pharmacological agents to inhibit TGF-beta-induced signaling and collagen deposition. Cs-A and IFN-gamma, but not glucocorticoids, cyclophosphamide, or azathioprine, inhibited TGF-beta-induced signaling, as assessed by luciferase reporter gene assays, and collagen deposition. TGF-beta antagonism by Cs-A was associated with direct inhibition of JunD activation, as demonstrated by electrophoretic mobility shift analyses. In contrast, the effects of IFN-gamma required signal transducer and activator of transcription (STAT)-1. We thus identify the JunD isoform of AP-1 as an essential mediator of TGF-beta-induced effects in lung fibroblasts. TGF-beta-induced signaling and collagen deposition are efficiently antagonized by Cs-A and IFN-gamma treatment, both of which exhibit distinct molecular mechanisms of action. These observations therefore offer novel targets for future therapy of fibrotic lung disease.
肺纤维化是一种致命疾病,其特征为细胞外基质(ECM)过度沉积,且与转化生长因子β(TGF-β)活性增加相关。尽管对其病理特征已有诸多了解,但我们对TGF-β刺激下导致ECM和胶原蛋白沉积增加的信号转导途径的认识仍不完全明确。我们之前报道过,转录因子AP-1的JunD同二聚体在肺成纤维细胞中被TGF-β特异性激活。在此我们证明,JunD也是TGF-β诱导效应所特别需要的。针对JunD而非c-fos或c-jun的反义寡核苷酸,能显著抑制原代人肺成纤维细胞中TGF-β诱导的胶原蛋白沉积。然后我们研究了药物抑制TGF-β诱导信号传导和胶原蛋白沉积的能力。通过荧光素酶报告基因检测评估,环孢素A(Cs-A)和γ干扰素(IFN-γ)而非糖皮质激素、环磷酰胺或硫唑嘌呤,能抑制TGF-β诱导的信号传导和胶原蛋白沉积。如电泳迁移率变动分析所示,Cs-A对TGF-β的拮抗作用与对JunD激活的直接抑制相关。相反,IFN-γ的作用需要信号转导及转录激活因子(STAT)-1。因此,我们确定AP-1的JunD异构体是肺成纤维细胞中TGF-β诱导效应的关键介质。Cs-A和IFN-γ处理能有效拮抗TGF-β诱导的信号传导和胶原蛋白沉积,二者均表现出独特的分子作用机制。因此,这些观察结果为未来纤维化肺病的治疗提供了新靶点。