GDNF基因修饰的的羊水干细胞干预肾间质纤维化的机制研究

慢性缺氧和小管间质性损伤是引起肾间质纤维化的主要因素，而肾小管周围毛细血管（PTC)损伤是引起肾脏缺氧和小管间质损伤的重要原因。申请者前期研究发现PTC损伤可以引起多种肾病模型肾间质纤维化，羊水干细胞 (AFS)干预可以改善PTC损伤和继发低氧，减轻肾间质纤维化程度。本项目拟在原有研究特色的基础上，应用单侧输尿管梗阻（UUO）小鼠模型，在整体、细胞和分子水平研究，重点探索胶质细胞源性神经营养因子（GDNF）基因修饰的AFS(GDNF-AFS)改善肾间质纤维化的机制。通过MAPKs通路蛋白的检测，了解低氧介导肾间质纤维化的信号通路。通过体外实验，观察GDNF-AFS向血管内皮样细胞和肾小管上皮细胞分化的能力。通过观察肾脏移植GDNF-AFS对UUO小鼠模型血管新生的影响和肾小管损伤的修复情况，研究GDNF-AFS干预肾间质纤维化的机制及可行性，为临床治疗肾脏纤维化提供新的思路。

Chronic hypoxia and tubulointerstitial injury is a major cause of renal interstitial fibrosis, and peritubular capillary (PTC) injury plays a critical role in renal hypoxia and tubulointerstitial injury. In our previous study, we found PTC injury was the important causes of several models of nephropathy; however, amniotic fluid derived stem cells (AFS) transplantation ameliorated PTC injury and hypoxia, and alleviated the progression of renal interstitial fibrosis. On the basis of previous studies, our project will focus on the protective mechanisms of glial cell line derived neurotrophic factor (GDNF) gene modified AFS(GDNF-AFS) on renal interstitial fibrosis in a murine model of unilateral ureteral obstruction (UUO). We will investigate the signal pathway of hypoxia-mediated renal interstitial fibrosis by examining the MAPKs pathway. We will observe the effect of GDNF-AFS on the differentiation of the endothelial-like cells and tubular epithelial cells by in vitro experiments. We will examine the effect of GDNF-AFS transplantation on angiogenesis and the restoration of tubular epithelial cells and investigate the mechanisms and the role of GDNF-AFS on the development of renal interstitial fibrosis. These results will provide insights on GDNF-AFS as a new cell source for the clinical treatment of renal fibrosis.