肾缺血再灌注损伤是缺血性急性肾功能衰竭的重要损伤环节,也是肾移植中影响移植肾早期功能恢复及长期存活的主要因素。PI3K/Akt激酶，ERK激酶及PKCε激酶等，在被激活的情况下可以显著提高组织细胞对缺血再灌注损伤的耐受作用。本课题基于钠钾ATP酶DR区特异性抗体通过诱导钠钾ATP酶活化而激活与细胞存活相关的细胞信号传导通路，深入研究PI3K/Akt，PKC ε及ERKs等蛋白激酶在肾缺血再灌注损伤中的保护作用，以及与组织中ROS水平，中性粒细胞浸润程度，各种血清炎症因子的表达水平的关系。本研究关注缺血性急性肾功能衰竭和肾移植后功能恢复的主要影响因素缺血再灌注损伤，从激活与细胞存活相关激酶的角度出发,提高组织细胞自身对缺血再灌注损伤的耐受，在理论上具有先进性，而且对于提高移植肾早期器官功能恢复效率及肾移植患者长期存活率具有重要理论价值和现实意义。

Renal ischemia reperfusion injury is an important injury part of ischemic acute renal failure, and it's also the main factors to affect the early functional recovery and the long-term survival of transplanted kidney in renal transplantation. Recently, It was reported that activated PI3K/AKT, ERK and PKCε could increase the tolerance of tissue to ischemia/reperfusion. It was also found that NKA (Na+/K+-ATPase) DR region specific antibody could induce cardioprotection against ischemic injury via Src/PI3K/Akt/ERK signal pathway. Based on these mechanism, we proposed that if DR region specific antibody (DRSAb) could also induce protective effect against ischemia/reperfusion injury on kidney. This proposal is therefore designed to: ..1).Comfirm if DRSAb could induce protective effect against ischemia on HK-2 cell line. Specifically, we will make and purify the DRSAb firstly. After that, HK-2 cells will be incubated with DRSAb or control. Cells were then subjected to ischemia. The viability of HK-2 cell will be measured 10 min after reperfusion by MTT assay...2).Investigate the signaling mechanisms underlying DRSAb induced protective effect.Specifically, we will examine the involvement of protein kinase C ε (PKCε), Extracellular signal-regulated kinases (ERK), and phosphoinositide-3 kinase (PI3K/Akt) pathways with western blot...3).Investigate the protect effect of DRSAb in kidney ischemia-reperfusion injury animal models. We will establish kidney ischemia/reperfusion injury animal models and examine the protective effect of DRSAb by detecting the inflammation factors and biochemistry index of serum after ischemia/reperfusion. We will also discuss the relationship among the activated protein kinase including PKCε, ERK, and PI3K/Akt pathways with western blot，the level of inflammation factors，such as TNF-a、IL-6、IL-18 and IL-17, the extent of neutrophile granulocyte infiltration and ROS level in tissue...This study focuses on the main factors of ischemia/reperfusion injury for functional recovery after ischemic acute renal failure and renal transplantation. Based on the point of cell survival kinase activation could improve the cell tolerance to ischemia/reperfusion injury, this project is advanced on theoretically, and has important theoretical and practical significance for improving the initial graft organ recovery efficiency and long-term survival of renal transplant patients.