缺血-再灌可诱导神经元出现程序性坏死，RIP1/RIP3介导的分子通路在其中发挥重要作用，RIP1磷酸化是启动该通路的核心要素之一。RIP1具有多个丝/苏氨酸磷酸化位点，但调控RIP1磷酸化的上游分子及作用具体位点仍不完全明确。Pin1可介导神经元死亡，其作用主要通过识别和调控底物蛋白丝/脯氨酸或苏/脯氨酸位点磷酸化实现。我们前期研究显示缺血-再灌诱导视网膜神经元Pin1/RIP1上调，两者间存在结合，Pin1可促进RIP1磷酸化；另外前期研究也显示CaMKII可调控Pin1，故我们推测CaMKII-Pin1-RIP1通路在缺血-再灌致神经元程序性坏死中发挥重要作用。我们拟通过分子生物学和细胞生物学等技术，首先探索Pin1识别和调控RIP1磷酸化位点及作用，再阐明CaMKII对于Pin1磷酸化及下游调控模式。本项目对揭示缺血-再灌致神经元程序性坏死的分子机制及具体调控靶点具有重要科学意义。

Ischemia-reperfusion (I/R) could induce neuronal necroptosis, which is tightly regulated by RIP1/RIP3 pathway. RIP1 phosphorylation is one of the key elements in this necrotic pathway. There are several serine/threonine phosphorylation sites in the RIP1. However, the RIP1 phosphorylation sites and its up-regulators are still not clearly investigated. Pin1 could mediate neuronal death by recognizing and regulating the phosphorylation of serine/proline or threonine/proline sites on substrate proteins. Our previous studies showed that I/R could increase the expression of Pin1/RIP1 in retinal neurons. Pin1 also could bind and promote the RIP1 phosphorylation. In addition, our studies showed that CaMKII could regulate the Pin1 activity. Taken together, we speculate that the CaMKII-Pin1-RIP1 pathway plays an important role in neuronal necroptosis induced by IR. In this study, we first aim to explore the phosphorylation sites of RIP1 recognized and regulated by Pin1 through different kinds of molecular and cellular biotechnologies. Then, we will clarify the regulatory role of CaMKII on Pin1 phosphorylation and down-regulatory mechanisms. This study will provide an important scientific significance for revealing the molecular mechanism and specific regulatory targets of neuronal necroptosis induced by I/R.