近年来程序性坏死研究取得重要进展，受体相互作用蛋白3（Receptor-interacting protein 3，RIP3）被发现是程序性坏死的分子开关。本课题组前期研究发现全脑缺血复灌损伤后神经元死亡特征符合程序性坏死，随之我们发现缺血损伤诱导RIP3在神经元核内表达显著升高，有别于肿瘤坏死因子诱导的程序性坏死，提示RIP3可能是调控神经元缺血性程序性坏死的关键分子，且靶位在细胞核内，该结果尚未见报道。本项目拟利用基因敲除小鼠，应用RNA干扰、基因转染、拮抗剂干预等方法；采用免疫共沉淀、免疫荧光双标、电镜等技术；从整体动物和细胞水平研究神经元缺血性程序性坏死时RIP3的变化特征及RIP3表达增高、核定位对神经元死亡的影响，阐明RIP3调控神经元程序性坏死的下游信号通路。该项目的实施有助于揭示RIP3在神经元缺血性程序性坏死中的作用及调控机制，有望为脑血管病治疗寻找新的有效干预靶点。

The studies about programmed necrosis made great progress recently, receptor-interacting protein 3 (RIP3) was found to be the molecular switch of programmed necrosis. Our recent studies showed that the changes of hippocampal cornu ammonis 1 neuronal death induced by global cerebral ischemia/reperfusion injury were programmed necrosis. And our following studies showed that the expression of RIP3 after ischemia/reperfusion injury increased greatly in neuronal nuclei, which has not been reported and was different with programmed necrosis induced by tumor necrosis factor. These suggested RIP3 may be the key regulator of neuronal programmed necrosis after global cerebral ischemia/reperfusion injury, and the regulated target may be in the nuclei. In this project, from the whole animal and cellular level, we try to study the changes of RIP3, and the effect of increased expression and nuclear location of RIP3 on neuronal death; and try to clarify the downstream signaling pathway of neuronal programmed necrosis regulated by RIP3. Molecular biology techniques and tools of RIP3 knock-out mouse, RNA interfering, gene transfection and expression, drug inhibition, co-immunoprecipitation, double immunofluorescent labeling and electron microscopy would be employed in this study. Implementation of this project is help to reveal the role and signal transduction mechanisms of RIP3 in neuronal ischemic programmed necrosis. This would provide a theoretical reference for cerebrovascular disease research, and find effective intervention target for cerebrovascular disease treatment.