脊髓缺血再灌注损伤是脊柱外科术后最严重的并发症之一。缺血预处理可减轻此种损伤，但具体机制不详。我们已证实Bcl-2是调控神经细胞死亡的重要蛋白（Fan J et al: 2010 Neurosci Lett; 2012 Int J Neurosci; 2013 Brain Res; 2013 Neurosci Lett; 2010 中国脊柱脊髓杂志），而缺血预处理可通过影响Bcl-2活性保护神经细胞。最新研究表明：Bcl-2亦是重要的自噬调节蛋白，且调节能力与其多位点磷酸化水平及亚细胞定位密切相关。本项目拟通过建立大鼠脊髓缺血再灌注损伤及脊髓神经体外缺氧缺糖再复氧复糖模型，运用免疫杂交等方法，探讨缺血预处理影响Bcl-2蛋白非结构性环状结构域内丝/苏氨酸磷酸化水平及亚细胞定位调节神经细胞自噬的具体机制。本研究将丰富对确缺血预处理具体保护机制的认识，为临床防治脊髓缺血再灌注损伤提供新思路。

Spinal cord ischemia-reperfusion injury (SCII) is one of the most severe complications after spinal surgery. Ischemic preconditioning could alleviate spinal cord ischemia-reperfusion injury, but the mechanism remains unclear. We proved that Bcl-2 play a role in neurons death（Fan J et al: 2010 Neuroscience Letters; 2012 Int J Neuroscience; 2013 Brain Research; 2013 Neuroscience Letters; 2010 Chinese Journal of Spine and Spinal Cord）, and Ischemic preconditioning could protect neurons by influencing the activity of Bcl-2. Recent studies showed that Bcl-2 plays an important role in autophagy regulation. In addition, the multisite phosphorylation and subcellular localization of Bcl-2 may be the key to regulate autophagy. In this project, we will explore the mechanism of Ischemic preconditioning-mediated the threonine and serine phosphorylation, which located in nonstructured loop domain, and subcellular localization of Bcl-2 regulate spinal nerve cells autophagy, by applying immunoblotting, immunoprecipitation, immunofluorescence in the rat model of SCII and oxygen-glucose-deprivation/ reoxygenation (OGD/R) injury in spinal nerves. If our experiments panned out, we can better understand the pathogenesis of Spinal cord ischemia-reperfusion (SCII), the protection mechanism of Ischemic preconditioning, and provide a new idea for SCII in clinical.