脂肪间充质干细胞上清液对小鼠皮肤创伤的作用及可能机制

The effect and possible mechanism of supernatant of adipose mesenchymal stem cells on the skin wound healing in mice

目的观察小鼠脂肪间充质干细胞(Adipose mesenchymal stem cells,ADSCs)上清液经尾静脉注射治疗小鼠皮肤创伤的效果并探讨其作用机制。方法无菌条件下分离培养正常小鼠的ADSCs取3-5代细胞通过免疫荧光法检测间充质干细胞相关的细胞表面标记物CD34、CD45、CD90、CD105的表达。建立小鼠全皮层皮肤创伤模型并随机分为2组,注射生理盐水的对照组和注射上清液的实验组,分别于伤后7天,14天观察创面愈合情况。分离小鼠成纤维细胞(Fibroblast,Fb),将小鼠ADSCs上清液作用于Fb不同时间后,应用WST法和Transwell迁移实验检测Fb细胞增殖和迁移情况,应用real-time PCR和Western blot检测Fb中碱性成纤维细胞生长因子(basci fibroblast growth factor,b FGF)在转录和蛋白水平的表达。结果小鼠ADSCs表达CD90和CD105,不表达CD34和CD45。ADSCs上清液注射组于创伤后第7天,14天的伤口愈合率分别为(65%±3.4%),(95.6%±5.2%),均显著高于7天、14天生理盐水对照组的(55%±4.4%),(77.1%±3.1%)。ADSCs上清液作用于Fb后,能促进Fb的增殖和迁移以及上调生长因子b FGF在转录和蛋白水平的表达。结论小鼠ADSCs上清液促进小鼠皮肤创面愈合可能与上调b FGF表达相关。

Objective To study the effect and possible mechanism of supernatant of adipose mesenchymal stem cells（ADSCs） on the skin wound healing in mice. Methods ADSCs from normal mice were harvested under sterile conditions, in which the expressions of CD34, CD45, CD90 and CD105 profiles were examined by immunofluorescence. The full-thickness skin wound mice models were established and randomly divided into saline control group and the supernatant group injected via tail vein.Then, the therapeutic potentials were compared at 7days and 14 days after injury.The proliferation and migration of fibroblast were determined by WST assay and transwell assay respectively. The expression of basic fibroblast growth factor（b FGF） was detected by real-time PCR and Western blot. Results ADSCs displayed fibroblast-like phenotype in shape, expressed stem cells markers positively, including CD90 and CD105, but not CD34 and CD45. At 7 days and 14 days after transplantation, wound healing rates were higher in ADSCs supernatant group than in control group. ADSCs supernatant promoted the proliferation and migration of fibroblast and upregulated b FGF expression in fibroblast significantly. Conclusion The promoted cutaneous wound healing of ADSCs supernatant might be related to the upregulation of b FGF expression in fibroblast.