摘要
目的研究缺血肌肉转染腺病毒(Ad)携带人血管内皮细胞生长因子165(VEGF-165)和人血管形成素-1(Ang-1)基因后引起的细胞动员效应。方法制作大鼠下肢血管闭塞模型,肌肉内注射Ad5VEGF-165和/或Ad5Ang-1,以Western法检测生长因子蛋白表达、免疫组化检测基因导入后在缺血肌肉引起的效应。结果(1)局部转基因肌肉组织高表达人VEGF-165蛋白和人Ang1蛋白;(2)与对照组相比,接收两因子转染后的骨骼肌纤维间溴化脱氧尿嘧啶(BrdU)阳性的浸润细胞显著增多,呈协同效应。有细胞同时表达内皮细胞、骨骼肌细胞、平滑肌细胞标志性抗原。部分细胞表达CD117+并分化为新生微血管的内皮细胞。结论(1)两因子能促进CD117+骨髓干细胞动员到缺血组织部位并分化为内皮细胞参与血管新生;(2)两因子可能协同动员召集能在缺血部位聚集并分化为其它类型细胞的干/祖细胞;(3)两因子可能同时作为血管生成因子和细胞动员剂用于缺血性疾病的干细胞移植治疗中,以增强缺血损伤修复效果和节省干细胞用量。
Objective To study the mobilization effect of stem cells of vascular endothelial growth factor 165 and angiopoietin-1(Ad-VEGF -165,Ad-Ang-1) in a rat model of hindlimb ischemia.Methods Ad-VEGF -165 and Ad-Ang-1 were intramuscularly transferred into the rat ischemic hindlimb and their protein expression was examined by Western blotting.Histochemical examination was performed to illustrate the effects on rat ischemic muscles after gene transferring.Results (1)Western blotting showed a high expression of VEGF -165 and Ang-1 in the ischemic hindlimb muscle transferred with Ad 5-VEGF 165 and Ad 5-Ang-1 VEGF -165 respectively.(2)Compared with the control groups,a singnificant increase in the number of infiltrating cells,most of which were positive for BrdU, was seen between the muscle fibers received VEGF -165 and/or Ang-1 injection,and the two cytokines had a synergic effect.Some of the cells were positive for the markers of endothelial cells,smooth muscle cells and skeletal muscle cells.Some were positive for CD117,the marker of bone marrow stem cells,and diferentiated to endothelial cells which were found incorporating to the new-formed microvasculars.Conclusion (1)VEGF -165 and Ang-1 can enhance the mobilization of CD117 + marrow stem cells to ischemic tissues where they differentiate to endothelial cells incorporating to the neowasularization.(2)Effects of gene therapy of VEGF -165 and Ang-1 were not limited to angiogenesis but may involve the mobilization and recruitment of marrow stem cell.(3)VEGF -165 and Ang-1 may be employed in the stem cell transplant therapy of ischemic diseases serving simultaniously as angiogenetic factors and cell mobilization factors,thus contribute to ischemic trauma rehabilitation and the saving of stem cells.
出处
《江苏医药》
CAS
CSCD
北大核心
2005年第5期360-362,F003,共4页
Jiangsu Medical Journal
基金
江苏省科委自然科学基金(BK2001118)
江苏省卫生厅科技项目(H2004)
