正常与糖尿病小鼠脂肪间充质干细胞移植促进皮肤创伤愈合的比较

目的比较正常和糖尿病小鼠脂肪间充质干细胞(Adipose mesenchymal stem cells,ADSCs)移植促进皮肤创伤愈合疗效差别。方法无菌条件下获取正常小鼠及糖尿病小鼠的ADSCs,应用流式细胞术对3代细胞表面抗原CD34、CD45、CD90、CD105进行表型鉴定。应用WST法和Transwell迁移实验检测细胞增殖和迁移情况。应用ELISA对正常小鼠及糖尿病小鼠的ADSCs条件培养液中VEGF、HGF和IGF-1蛋白含量进行检测。建立小鼠皮肤创伤模型并随机分为3组,分别以皮内注射方式将3代正常小鼠或糖尿病小鼠来源ADSCs的细胞混悬液移植到小鼠创面四周,空白对照组注射生理盐水,于伤后14d观察创面愈合情况;Western blot检测伤后14d创面组织Bcl-2蛋白表达。结果小鼠ADSCs表达CD90和CD105,不表达CD34和CD45,与正常小鼠相比,糖尿病小鼠来源的ADSCs增殖和迁移能力下降,条件培养液中VEGF、HGF和IGF-1蛋白含量明显降低。糖尿病小鼠ADSCs移植组于创伤后第14d伤口愈合率为(79.6%±6.2%),均显著低于正常小鼠移植组14d的(97.1%±4.1%),两者均高于对照组14d的(64.6%±2.9%,0.05)。与正常小鼠移植组相比,糖尿病小鼠移植组伤后14d创面组织Bcl-2表达水平降低,高于对照组创面组织Bcl-2表达水平。结论正常小鼠来源ADSCs较糖尿病小鼠ADSCs更能促进小鼠皮肤创面愈合。

Adipose mesenchymal stem cells from normal versus diabetic rats for promoting cutaneous wound healing

Objective To compare the therapeutic effects of adipose mesenchymal stem cells （ADSCs） from normal and diabetic mice on cutaneous wound healing. Methods ADSCs from normal and diabetic mice were harvested under sterile conditions. The expression profiles of CD34, CD45, CD90 and CDI05 were examined by immunofluorescenee. The proliferation and migration of ADSCs were determined by WST assay and transwell assay respectively. In addition, the contents of vascular endothelial growth factor（VEGF）, hepatocyte growth factor（HGF） and insulin-like growth factor-l（IGF- 1） were determined by ELISA analysis. Then, the therapeutic potentials were compared by transplanting equal numbers of normal or diabetic ADSCs in a mouse model of cutaneous wounds. Bcl-2 expression was detected by Western blot in wound areas at 14 days after damage. Results The morphological feature of ADSCs displayed fibroblast-like phenotype. The cells were positive for stem cells markers, including CD90 and CD105, and negative for CD34 and CD45 by addition, compared with normal group, the proliferation and migration were significantly inhibited in ADSCs from dibatic mice. At 14 days after transplantation, wound healing rate was higher in normal ADSCs （97.1% ± 4.1%） than in diabetic groups （79.6% ± 6.2%）, but both higher than in empty control group （64.6% ± 2.9%）. Bcl-2 expression level in the wound area was higher in normal group than in diabetic group. Conclusion The ADSCs from normal rats have better therapeutic effects on cutaneous wound healing than those from diabetic rats.