Heat Shock Protein 90 Inhibitor Decreases Collagen Synthesis of Keloid Fibroblasts and Attenuates the Extracellular Matrix on the Keloid Spheroid Model.
Heat Shock Protein 90 Inhibitor Decreases Collagen Synthesis of Keloid Fibroblasts and Attenuates the Extracellular Matrix on the Keloid Spheroid Model.
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热休克蛋白90抑制剂降低了乳头状成纤维细胞的胶原蛋白合成，并减轻了酮球体模型上的细胞外基质。
DOI：
10.1097/prs.0000000000001538
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发表时间：
2015-09
影响因子：
3.6
通讯作者：
Yun CO
中科院分区：
文献类型：
--
作者：
Lee WJ;Lee JH;Ahn HM;Song SY;Kim YO;Lew DH;Yun CO
热休克蛋白90（HSP 90）是一种丰富的细胞质伴侣蛋白，17-烯丙基氨基-17-去甲氧基格尔德霉素（17-AAG）抑制HSP 90可通过促进转化生长因子（TGF）-β受体I和II的降解，从而抑制Smad 2/3的激活，从而降低TGF-β介导的转录应答。在此，我们评估了HSP 90是否在瘢痕疙瘩的发病机制和治疗中调节TGF-β信号传导。用17-AAG（10 μM）处理瘢痕疙瘩成纤维细胞，并通过实时RT-PCR测定I型和III型胶原的mRNA水平。采用酶联免疫吸附试验（ELISA）检测TGF-β1的表达。Western blot检测17-AAG对Smad 2/3复合物蛋白水平的影响。此外，在17-AAG处理的瘢痕疙瘩球体中，胶原沉积和主要细胞外基质蛋白的表达通过Masson三色染色和免疫组织化学进行了研究。我们发现，与正常组织相比，瘢痕疙瘩组织中HSP 90表达过强，17-AAG可降低瘢痕疙瘩成纤维细胞中I型胶原、分泌型TGF-β1、Smad 2/3复合物蛋白的表达。Masson三色染色显示17-AAG处理的瘢痕疙瘩球体中胶原沉积减少，免疫组化分析显示17-AAG处理的瘢痕疙瘩球体中胶原I和III、弹性蛋白和纤连蛋白的表达显著减少。这些结果表明，HSP 90抑制剂如17-AAG的抗纤维化作用可能对瘢痕疙瘩有治疗作用。
The 90-kDa heat-shock protein (HSP90) is an abundant cytosolic chaperone and inhibition of HSP90 by 17-allylamino-17-demethoxygeldanamycin (17-AAG) compromises Transforming growth factor (TGF)-β-mediated transcriptional responses by enhancing TGF-β receptor I and II degradation, thus preventing Smad 2/3 activation. Here, we evaluated whether HSP90 regulates TGF-β signaling in the pathogenesis and treatment of keloids. Keloid fibroblasts were treated with 17-AAG (10 μM), and mRNA levels of collagen type I and III were determined by real-time RT-PCR. Also, secreted TGF-β1 was assessed by Enzyme-linked immunosorbent assay (ELISA). The effect of 17-AAG on protein levels of Smad 2/3 complex was determined by Western blot. Additionally, in 17-AAG-treated keloid spheroids, the collagen deposition and expression of major extracellular matrix proteins were investigated by Masson's trichrome staining and immunohistochemistry. We found that HSP90 is overexpressed in human keloid tissue compared to adjacent normal tissue, and 17-AAG decreased mRNA levels of type I collagen, secreted TGF-β1, Smad 2/3 complex protein expression in keloid fibroblasts. Masson's trichrome staining revealed that collagen deposition was decreased in 17-AAG-treated keloid spheroids, and immunohistochemical analysis showed that expression of collagen I and III, elastin, and fibronectin were markedly decreased in 17-AAG-treated keloid spheroids. These results suggest that the antifibrotic action of HSP90 inhibitors such as 17-AAG may have therapeutic effects on keloids.