摘要
目的评价骨髓间充质干细胞(BM-MSCs)经小腿肌肉移植对大鼠糖尿病足溃疡(DFUs)的治疗效果。方法全骨髓贴壁法体外培养雄性Wistar大鼠BM-MSCs至第3代,经4,6-联脒-2-苯基吲哚(DAPI)标记。36只雄性Wistar大鼠随机分成A组(BM-MSCs小腿肌肉移植组,n=12)、B组(正常足溃疡对照组,n=12)和C组(DFUs对照组,n=12)。大鼠糖尿病经腹腔注射链脲佐菌素(STZ)诱导,然后在其双后足背上切取3 mm×7 mm矩形全层皮肤制成大鼠DFUs模型。于移植后第2、5、8、11天评估各组大鼠的创面愈合率,冰冻切片观察DAPI标记的BM-MSCs在创伤组织中的示踪,肉芽组织的厚度采用HE染色法检测,CD31和Ki-67的表达采用免疫组化法检测,血管内皮细胞生长因子(VEGF)在创伤组织中的表达水平采用ELISA和RT-PCR检测。结果 B组的创面愈合最快(P<0.05),第8、11天A组的创面愈合率高于C组(P<0.05)。冰冻切片发现A组荧光强度和区域在第5天时达到高峰。HE染色结果显示第5天A、B两组的肉芽组织厚度相同,但比C组厚。CD31和Ki-67的免疫组化发现A、B两组的平均阳性小血管数和细胞数无差异,但与C组相比有差异(P<0.05)。ELISA和RT-PCR结果均显示,第11天A组的VEGF表达水平明显较对照组高(P<0.001)。结论 BM-MSCs经小腿肌肉移植能明显促进大鼠DFUs的愈合,这种促愈疗效可能是通过促进创伤组织中VEGF的表达实现的。
Objective To assess the therapeutic effect of intramuscular transplantation of bone-marrow derived mesenchymal stem cells （BM-MSCs） in the leg for treatment of diabetic foot ulcers （DFUs） in rats. Methods Thirty-six male Wistar rats were randomly allocated into 3 equal groups, namely group A with DFUs on the bilateral hindlimb dorsum pedis and intramuscular transplantation of 4,6-diamino-2-phenylindole （DAPI）-labeled third-passage BM-MSCs from male Wistar rats into the leg, group B with nondiabetic foot ulcers, and group C with DFUs but without BM-MSC transplantation. On days 2, 5, 8 and 11 posttransplantation, the rate of wound healing was evaluated, the labeled BM-MSCs in the wound tissues were traced on frozen sections, and the thickness of granulation tissues was measured with HE staining. Immunohistochemistry was performed to detect the expression of CD31 and Ki-67, and the expression of vascular endothelial growth factor （VEGF） in the wound tissues was detected by ELISA and RT-PCR. Results The rats in group B showed significantly faster wound closure than those in the other two groups （P〈0.05）. The rate of wound healing was greater in group A than in group C on days 8 and 11 （P〈0.05）. The fluorescence intensity and area on the frozen sections were the highest on day 5 posttransplantation in group A, which had a granulation tissue thickness comparable with group B but greater than group C. The mean numbers of small blood vessels and cells positive for CD31 and Ki-67 expressions were similar between groups A and B, which showed significant differences from group C （P〈0.05）. On day 11, group A showed the highest VEGF expression in the wound tissues （P〈0.001）. Conclusion Intramuscular transplantation of BM-MSCs can significantly promote wound healing of DFUs in rats possibly as a result of increased expression of VEGF in the wound tissues.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2012年第12期1730-1736,共7页
Journal of Southern Medical University
基金
兰州军区面上项目(CLZ11JB07)
关键词
骨髓间充质干细胞
糖尿病足
创面修复
bone-marrow derived mesenchymal stem cells
diabetic foot
wound healing
