Functional characterization of TRAP1-like protein involved in modulating fibrotic processes mediated by TGF-β/Smad signaling in hypertrophic scar fibroblasts.
Functional characterization of TRAP1-like protein involved in modulating fibrotic processes mediated by TGF-β/Smad signaling in hypertrophic scar fibroblasts.
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DOI：
10.1016/j.yexcr.2015.01.015
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发表时间：
2015-03
影响因子：
3.7
通讯作者：
Xue Wang;J. Chu;Congji Wen;Shuiting Fu;Yun-liang Qian;Yan Wo;Chuandong Wang;Danru Wang
中科院分区：
文献类型：
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作者：
Xue Wang;J. Chu;Congji Wen;Shuiting Fu;Yun-liang Qian;Yan Wo;Chuandong Wang;Danru Wang
转化生长因子-β-1(转化生长因子-TLP)介导的信号通路与创面愈合和瘢痕形成密切相关，其中β样蛋白在调节Smad2/Smad3信号平衡中起着重要作用。我们先前的研究揭示了正常皮肤成纤维细胞中TLP与胶原合成的关系。在此，我们将详细分析TLP在增生性瘢痕形成和收缩过程中的作用。为了探讨和验证TLP在增生性瘢痕成纤维细胞(HSFb)发病机制中的作用，我们构建了高表达TLP或编码针对TLP的小发夹RNA(ShRNAs)的慢病毒载体，并将其导入HSFb。与对照组相比，HSFb中TLP基因敲除导致细胞收缩、I型和III型胶原mRNA转录和蛋白表达水平降低，纤维连接蛋白(FN)水平升高。此外，与对照组相比，TLP基因敲除可促进人增生性瘢痕成纤维细胞Smad3的磷酸化，抑制Smad2和Erk-1/2的磷酸化。TLP的减少并不影响HSF的增殖能力，但外源性TLP与转化生长因子-β-1协同作用可提高细胞存活率。综上所述，我们的发现证明了TLP表达在增生性瘢痕形成和收缩中的作用。
The transforming growth factor-β1 (TGF-β)-mediated signaling pathway is believed to be closely associated with wound healing and scar formation, in which TRAP1-like protein (TLP) plays a role in regulating the balance of Smad2 vs. Smad3 signaling. Our previous study revealed the relation between TLP and collagen synthesis in normal human skin fibroblasts. Here, we present a detailed analysis of the effects of TLP on the process of hypertrophic scar formation and contraction. To explore and verify a contribution of TLP to the pathological mechanism of hypertrophic scar fibroblasts (HSFb), we constructed lentiviral vectors that either overexpressed TLP or encoded small hairpin RNAs (shRNAs) targeting TLP, then we transfected them into HSFb. TLP knockdown in HSFb resulted in reduced levels of cell contraction, type I and type III collagen mRNA transcripts and protein expression, and higher levels of fibronectin (FN) compared to control groups. In addition, knockdown of TLP promoted the phosphorylation of Smad3 but repressed Smad2 and Erk-1/2 phosphorylation in human hypertrophic scar fibroblasts compared to control groups. The reduction of TLP did not interfere with HSF proliferative ability, but exogenous TLP cooperated with TGF-β1 to increase cell viability. Together, our findings demonstrate evidence for a contribution of TLP expression in hypertrophic scar formation and contraction.