神经损伤是脑缺血再灌注（I/R）损伤的重要组成部分。我们预实验结果提示: I/R后神经元自噬流破坏，Ykt6与SEC22b可能参与其中。为此，我们提出假说：Ykt6与SEC22b参与神经元自噬流，I/R后二者表达失衡，导致神经元自噬流紊乱，加剧脑损伤。为了验证该假说，我们拟利用大鼠I/R模型，应用电镜和western blot等检测神经元自噬流以及Ykt6与SEC22b的水平；应用基因调控和转基因，评估调控Ykt6与SEC22b表达的关键因子以及二者的表达变化对神经元自噬流完整性和神经损伤的影响；利用自噬流抑制剂及自噬缺失小鼠阻断自噬，明确Ykt6与SEC22b是否通过调控自噬流发挥作用；利用免疫荧光染色等技术，明确Ykt6与SEC22b调控神经元自噬流的机制。本研究纵向从细胞、组织和活体三个层面，横向从超微结构，分子生物学和行为学三个层面研究，将为改善脑I/R 预后提供新思路和理论依据。

Neuronal injury is an important part of cerebral ischemia-reperfusion (I/R) injury. Our preliminary experimental results showed that I/R induced autophagic flux damage and that Ykt6 and SEC22b were probably involved in the process. Thus, we proposed the hypothesis: Ykt6 and SEC22b mediate the integrity of autophagic flux in neurons. After I/R, the expression of Ykt6 and SEC22b is disturbed, which promotes autophagic flux damage, and finally brain injury. To test the hypothesis, we use experimental I/R model in rats. First, we discover the effect of I/R on autophagic flux and the expression of Ykt6 and SEC22b in neurons of the penumbral tissue. Second, gene transfection, RNAi and transgenic mice are used to regulate the target gene expression, and then to evaluate the role of Ykt6 and SEC22b in I/R-induced autophagic flux damage and brain injury. Third, autophagy-blocking agent and autophagy-deficient mice are used to test whether Ykt6 and SEC22b exert their effects via regulating autophagy. And finally, by double immunofluorescence, we demonstrate the mechanisms underlying the effect of Ykt6 and SEC22b on I/R-induced autophagic flux damage and brain injury. The longitudinal studies from three levels of cell, tissue and in vivo, horizontal researches from three levels of ultrastructure, molecular biology and behavior in this study will provide us new ideas and theoretical basis to improve the prognosis of ischemic stroke patients.