Woeller Collynn F, O'Loughlin Charles W, Roztocil Elisa, Feldon Steven E, Phipps Richard P
From the Department of Environmental Medicine and.
Flaum Eye Institute, School of Medicine and Dentistry, University of Rochester, Rochester, New York 14642.
J Biol Chem. 2015 Feb 6;290(6):3563-75. doi: 10.1074/jbc.M114.601872. Epub 2014 Dec 23.
Although scarring is a component of wound healing, excessive scar formation is a debilitating condition that results in pain, loss of tissue function, and even death. Many tissues, including the lungs, heart, skin, and eyes, can develop excessive scar tissue as a result of tissue injury, chronic inflammation, or autoimmune disease. Unfortunately, there are few, if any, effective treatments to prevent excess scarring, and new treatment strategies are needed. Using HEK293FT cells stably transfected with a TGFβ-dependent luciferase reporter, we performed a small molecule screen to identify novel compounds with antiscarring activity. We discovered that the polyether ionophore salinomycin potently inhibited the formation of scar-forming myofibroblasts. Salinomycin (250 nm) blocked TGFβ-dependent expression of the cardinal myofibroblast products α smooth muscle actin, calponin, and collagen in primary human fibroblasts without causing cell death. Salinomycin blocked phosphorylation and activation of TAK1 and p38, two proteins fundamentally involved in signaling myofibroblast and scar formation. Expression of constitutively active mitogen activated kinase kinase 6, which activates p38 MAPK, attenuated the ability of salinomycin to block myofibroblast formation, demonstrating that salinomycin targets the p38 kinase pathway to disrupt TGFβ signaling. These data identify salinomycin and other polyether ionophores as novel potential antiscarring therapeutics.
尽管瘢痕形成是伤口愈合的一个组成部分,但过度的瘢痕形成却是一种使人衰弱的病症,会导致疼痛、组织功能丧失甚至死亡。包括肺、心脏、皮肤和眼睛在内的许多组织,都可能因组织损伤、慢性炎症或自身免疫性疾病而形成过多的瘢痕组织。不幸的是,几乎没有有效的治疗方法来预防过度瘢痕形成,因此需要新的治疗策略。我们利用稳定转染了TGFβ依赖性荧光素酶报告基因的HEK293FT细胞,进行了小分子筛选,以鉴定具有抗瘢痕活性的新型化合物。我们发现聚醚离子载体盐霉素能有效抑制瘢痕形成性肌成纤维细胞的形成。盐霉素(250纳米)可阻断原代人成纤维细胞中主要肌成纤维细胞产物α平滑肌肌动蛋白、钙调蛋白和胶原蛋白的TGFβ依赖性表达,且不会导致细胞死亡。盐霉素可阻断TAK1和p38的磷酸化及激活,这两种蛋白是肌成纤维细胞信号传导和瘢痕形成的根本参与者。组成型活性丝裂原活化激酶激酶6(其激活p38 MAPK)的表达减弱了盐霉素阻断肌成纤维细胞形成的能力,这表明盐霉素靶向p38激酶途径来破坏TGFβ信号传导。这些数据表明盐霉素和其他聚醚离子载体是新型潜在的抗瘢痕治疗药物。