摘要
本研究观察转染血管内皮生长因子165基因的骨髓间充质干细胞(MSC)移植对大鼠缺血坏死下肢的治疗作用。选择成年SD大鼠,在无菌条件下分离右股动脉及其分支后分别结扎,建立下肢缺血坏死模型。术后第7天,将DAPI标记的转染了重组腺病毒装载的人vegf-165基因(ad-vegf-165)的MSC(MSC-vegf组)及转染携带半乳糖苷酶基因的重组腺病毒(Ad-LacZ)的MSC(MSC-LacZ组)及不处理的MSC(MSC组),分别局部注射到缺血坏死的下肢骨骼肌中。用Western blot法分析MSC及其骨骼肌组织pAkt和VEGF蛋白的表达。注射后1、2、4周,用免疫组织化学法观察在缺血坏死下肢组织中MSC的FⅧ和肌球蛋白的表达。结果表明:在荧光显微镜下各组注射部位均可见移植的MSC,并且在MSC-vegf组MSC细胞数目明显多于其它组。Western blot分析发现,MSC-vegf组呈现出高水平的pAkt表达及高水平的VEGF蛋白表达。MSC-vegf组的MSC于移植后1、2、4周呈现出FⅧ阳性特征,且阳性率显著高于MSC-LacZ组和MSC组;而肌球蛋白阳性率,在MSC-vegf组、MSC-LacZ组和MSC组之间无显著差异。结论:转染vegf的MSC通过显著提高缺血坏死部位VEGF的表达量促进血管新生,修复缺血坏死下肢。
The aim of study was to explore the effect of mesenchymal stem cells （MSC） transfected with recombinant adenovirus-mediated human vascular endothelium growth factor 165 （ad-vegf-165） on treating ischemic necrosis limbs. Adult SD rats were selected for study. Limb ischemic necrosis model was established by right femoral artery ligation in SD rats. 7 days after ligation, MSC, ad-h-vegf-165-MSC and ad-LacZ-MSC labelled by DAPI were injected into ischemic necrosis limb in rats respectively. One week after injection, the expression of VEGF in ischemic necrosis limbs was detected by Western blot. And at 1, 2 or 4 weeks after injection, the expressions of FⅧ and myosin on MSC were evaluated by immunohistochemistry. The results indicated that the MSC labelled by DAPI could be found in the transplantation site of ischemic necrosis limbs under fluorescent microscope. And the number of MSC in MSC-vegf group was more than that in MSC and MSC-LacZ groups. The VEGF expression in MSC-vegf group was higher than that in MSC and MSC-LacZ groups. More importantly, the number of endothelial cells demonstrated characteristic FⅧ positive MSC in MSC-vegf group was more than that in MSC and MSC-LacZ group after injections of 1, 2 and 4 weeks. However, the number of myosin positive MSC among MSC-vegf, MSC and MSC-LacZ groups showed no significant difference. It is concluded that MSC transfected with Ad-vegf promotes angiogenesis to repair ischemic necrosis limbs through the increased expression of VEGF.
出处
《中国实验血液学杂志》
CAS
CSCD
2010年第6期1568-1573,共6页
Journal of Experimental Hematology
