缝隙连接传递活性氧通过PIAS3负调控STAT3在肝移植急性肾损伤修复中的作用研究

Kidney has strong potential to repair, but acute kidney injury (AKI)following orthotropic liver transplantation (OLT)delays or can't repair, its mechanism is unclear.STAT3 is an important nuclear transcription factor to start and promote repair process; our previous study have found that low STAT3 expression in renal tissue after OLT，which suggested some factors restricting renal repair. The level of Reactive oxygen species (ROS) determine its biological efficiency and ROS activation PIAS3 is an important factor restricting the role of STAT3.However, the regulatory mechanism of ROS level is undefined. Gap junction (GJ) is intercellular membrane channel that mediate direct communication between neighboring cells and our previous study have found that inhibition of the GJ reduced kidney ROS levels following OLT. Thus, we assume GJ mediate ROS inhibiting AKI repairs following OLT through negative regulation of STAT3 via PIAS3.In vivo and in vitro studies, various methods, such as specific inhibitor, gene silencing and Cx32 gene knockout mice, will be applied to change the function of GJ, and indicators including renal function and pathological, PIAS3/STAT3 signal related protein and ROS transmitting will be detected to verify the hypothesis. The proposed study will not only provide new mechanistic insight in AKI repair, but also provide experimental and practical guidance for the development of effective therapeutic strategies combating against AKI following OLT.

肾脏虽然具有强大修复潜能，但肝移植急性肾损伤（AKI）普遍修复延迟或不能修复，其机制不明。STAT3是促进损伤修复的重要核转录因子，我们前期研究发现肝移植肾组织STAT3低表达，提示存在制约修复的因素；活性氧（ROS）水平决定其生物效能，最新研究发现ROS通过PIAS3抑制STAT3作用，但调控ROS水平的机制仍未明确。缝隙连接（GJ）是细胞间直接联系通道，我们前期研究发现抑制GJ减少肝移植肾组织ROS水平。据此，我们设想GJ传递的ROS通过PIAS3负调控STAT3信号抑制肝移植AKI的修复。本课题拟用特异性抑制剂、基因沉默和质粒转染、基因敲除鼠等技术改变GJ功能，通过肝移植动物实验和体外细胞学研究，检测肾功能和病理、PIAS3/STAT3信号相关蛋白以及ROS传递等指标以验证假说，进一步阐明肝移植AKI修复机制，为肝移植AKI早期修复治疗提供理论及实验依据。