机械创伤诱导皮肤成纤维细胞去分化的实验研究  

Experimental studies on dedifferentiation of skin fibroblasts induced by mechanical trauma

作　　者：陈泽林[1] 史春梦[1] 

机构地区：[1]第三军医大学军事预防医学院全军复合伤研究所,创伤、烧伤与复合伤国家重点实验室,重庆市纳米医药工程技术研究中心,重庆400038

出　　处：《感染．炎症．修复》2017年第2期84-89,129,130,共8页Infection Inflammation Repair

基　　金：国家973计划基金资助项目(2012CB518103)

摘　　要：目的:研究机械创伤是否能诱导皮肤成纤维细胞发生去分化。方法:建立体外培养细胞划痕损伤模型和体内全层皮肤损伤模型,应用免疫荧光染色、免疫组织化学、蛋白质印迹、转录组测序、si RNA等方法,检测机械创伤后成纤维细胞的形态、增殖情况、干细胞相关标志物的表达、集落形成能力、表达谱等的变化。结果:机械创伤后成纤维细胞逐渐变小变圆,发生类干细胞形态转变,并于创伤后36 h被激活而发生明显增殖,且创伤后成纤维细胞的激活不受创伤严重程度的影响。机械创伤后成纤维细胞中的干细胞标志物Sox2、波形蛋白的表达明显增强,集落形成能力也明显提高。体内实验也发现创面肉芽组织中出现大量Sox2阳性细胞。表达谱分析发现成纤维细胞的多能干细胞通路及相关调控分子表达明显增强。机械划痕后2 h创缘成纤维细胞中的总β-catenin和ser 675磷酸化的β-catenin的表达较对照组明显增强;应用β-catenin si RNA抑制成纤维细胞内的β-catenin表达后,机械创伤后创缘成纤维细胞中的Sox2阳性细胞几乎消失。结论:机械创伤可诱导皮肤成纤维细胞的去分化;β-catenin介导了机械创伤后成纤维细胞的去分化过程。Objective： To study whether mechanical trauma could induce dedifferentiation of skin fibroblasts. Methods： The scratch wound model of cells cultured in vitro and the full-thickness skin injury model in vi^vo were established, and the methodologies of immunofluorescence staining, immunohistochemistry, Western bloting and RNA sequencing,siRNA etc. were employed to detect the morphological change, proliferation, expression of stem cell related markers, colony forming ability and gene expression profile of fibroblasts. Results： Following mechanical trauma, fibroblasts changed themselves into round-small shapes, and were activated to proliferation within 36 hours. The activation was not dependent on the wound severity. The expressions of stem cell markers Sox2 and vimentin and the colony forming ability were dramatically increased following mechanical trauma. There also found a large number of Sox2＋ cells in the granulation tissues in the full-thickness wound model. Analysis of gene expression profile showed that the signaling pathway regulating pluripotency of stem cell and related regulating genes were significantly up-regulated. p-catenin and ser 675-phosphorylated p-catenin expressed by fibroblast enhanced 2 hours after mechanical trauma, but the Sox2 positive cells almost disappeared post-injury after p-catenin was inhibited by p-catenin siRNA. Conclusions： Mechanical trauma may induce dedifferentiation of skin fibroblasts, and p-catenin has been found to mediate the mechanical trauma induced dedifferentiation.

关 键 词：成纤维细胞 去分化 机械创伤 

分 类 号：R622.1[医药卫生—整形外科]