TGF-β1诱导表皮干细胞向间质细胞转化对增生性瘢痕形成的影响

摘要

目的：探讨转化生长因子-β1（Transform growth factor-β1，TGF-β1）对人表皮干细胞（Human epidermal stem cell， hESCs）向间质细胞转化的影响及该现象对瘢痕增生的作用。方法：体外原代培养hESCs，分析细胞表面标志物表达，不同浓度TGF-β1刺激后，MTT 法检测细胞增殖活性，酶联免疫吸附法（ELISA）检测上皮-间质细胞转化（Epithelial-mesenchymal transformation，EMT）正相关蛋白：波形蛋白（Vimentin，Vim）、Smad3，负相关蛋白：E-钙粘着蛋白（E-cadherin，E-cad）。建立兔耳瘢痕模型，不同浓度TGF-β1干预，术后4周取材行组织形态学观察。结果：利用人包皮皮肤成功培养出hESCs，表面标记物表达阳性。不同浓度TGF-β1可促进hESCs增殖，使Vim、Smad3表达上调，E-cad表达下调，且在一定浓度范围呈剂量依赖，兔耳瘢痕标本显示各组均有瘢痕增生，TGF-β1刺激组更为明显。结论：hESCs可发生EMT现象，该现象与瘢痕增生程度一致并可被TGF-β1诱导。推测瘢痕增生的实质可能为EMT。%Objective: To explore the effect of EMT of hESCs induced by TGF-β1,and the relationship with scar hyperplasia.Methods:hESCs were primarily cultured from human foreskin,treated with different dose of TGF-β1.We detect the expression of cell surface maker,MTT and ELISA were used to detect the proliferation of hESCs and the expression of Vim、Smad3、E-cad,which were positive and negative factors of EMT respectively. Rabbit ear scar model were established and intervened with different dose of TGF-β1, the scar were obtained after 4 weeks,and were observed on histomorphology. Results:hESCs were isolated and cultured successfully, the specific marker were expressed,TGF-β1 promoted proliferation of hESCs and expression of Vim、Smad3,reduced E-cad,the changes was dose-dependent in a certain range of concentration. Animal experiment indicated that scar hyperplasia were more marked in TGF-β1 group comparing with control group. Conclusion:EMT phenomenon exists in hESCs which was consistent with the degree of scar hyperplasia,and it can be induced by TGF-β1.