VEGF165基因转染的内皮祖细胞移植对大鼠肾脏缺血再灌注后的保护作用

评估VEGF165基因转染的内源性内皮祖细胞(EPCs)移植治疗肾脏缺血再灌注损伤(IRI)的作用，进一步完善EPCs对肾脏IRI产生保护作用的旁分泌机制。利用重组腺病毒载体Ad-VEGF165感染EPCs，并进行转染效率鉴定。并进一步移植治疗大鼠肾脏IRI，术后24h和72h分别评估血清肌酐及尿素氮水平；组织病理学检查评估各组大鼠术后肾损伤程度；Western blot检测大鼠肾脏IRI中血管内皮生长因子 (VEGF)、血管生成素-1 (Ang-1)及血管生成素-2 (Ang-2) 表达情况。结果发现利用重组腺病毒载体Ad-VEGF165转染的EPCs移植治疗后，大鼠肾脏功能明显改善，术后72h检测大鼠患肾，其内VEGF、Ang-1以及Ang-2表达水平均明显提升。因此，VEGF165基因转染的EPCs移植治疗可以有效的治疗大鼠肾脏IRI，其作用机制可能与EPCs归巢后旁分泌过量VEGF并进一步促进Ang-1、Ang-2等血管新生因子表达，从而促使肾脏血管新生有关。

VEGF mediates endothelial progenitor cells homing in the rat model of renal ischemia and reperfusion injury

The aim of this study was to assess whether vascular endothelial growth factor (VEGF) mediated the mobilization and recruitment of endothelial progenitor cells (EPCs) to protect kidneys from ischemia/reperfusion injury (IRI) in male rats. Recombinant adenovirus encoding VEGF165 gene (Ad-VEGF165) was used to infect EPCs, and the transfection efficiency was identified. Male Sprague-Dawley rats were injected EPCs suspension liquid (1×106) which was transfected by Ad-VEGF165 from the femoral vein of rats after the operation method of IRI-operated group. At 24 h and 72 h after reperfusion, serum samples were respectively collected for renal function. Besides, kidney tissues were harvested to observe renal morphology changes. Subsequently, vascular endothelial growth factor (VEGF), angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2) expression levels in the kidneys were measured using quantitative real-time PCR and western blot analysis at the indicated time points after reperfusion. The results found that the renal function was significantly improved, after EPCs transplantation with Ad-VEGF165. At 72 hours after reperfusion, expression levels of VEGF, Ang-1 and Ang-2 in the kidneys of EPCs-treated rats which was transfected by Ad-VEGF165 were markedly increased compared to rats subjected to IRI. Therefore, the present work suggested that EPCs transplantation with Ad-VEGF165 could effectively treat kidney IRI in rats. In addition, VEGF might play important roles in the protective effect of homing of EPCs on renal acute IRI, to further promote the expression of Ang-1, Ang-2 and other angiogenic factors.