Wang X, Chu J, Wen C J, Fu S B, Qian Y L, Wo Y, Wang C, Wang D R
Department of Plastic and Reconstructive Surgery, Shanghai Ninth People's Hospital, Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China; Department of Pediatric Surgery, Shanghai Children's Medical Center, Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China.
Department of Pediatric Surgery, Shanghai Children's Medical Center, Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China.
Exp Cell Res. 2015 Mar 15;332(2):202-11. doi: 10.1016/j.yexcr.2015.01.015. Epub 2015 Feb 2.
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.
转化生长因子-β1（TGF-β）介导的信号通路被认为与伤口愈合和瘢痕形成密切相关，其中TRAP1样蛋白（TLP）在调节Smad2与Smad3信号平衡中发挥作用。我们之前的研究揭示了TLP与正常人皮肤成纤维细胞中胶原蛋白合成之间的关系。在此，我们详细分析了TLP对肥厚性瘢痕形成和收缩过程的影响。为了探索和验证TLP对肥厚性瘢痕成纤维细胞（HSFb）病理机制的作用，我们构建了过表达TLP或编码靶向TLP的小发夹RNA（shRNA）的慢病毒载体，然后将它们转染到HSFb中。与对照组相比，HSFb中TLP的敲低导致细胞收缩水平降低、I型和III型胶原蛋白mRNA转录本及蛋白表达水平降低，以及纤连蛋白（FN）水平升高。此外，与对照组相比，TLP的敲低促进了人肥厚性瘢痕成纤维细胞中Smad3的磷酸化，但抑制了Smad2和Erk-1/2的磷酸化。TLP的减少并未干扰HSF的增殖能力，但外源性TLP与TGF-β1协同作用可增加细胞活力。总之，我们的研究结果证明了TLP表达在肥厚性瘢痕形成和收缩中的作用。
