SNAP29通过调控自噬溶酶体形成介导高压氧预处理脑保护效应的分子机制研究

The prevention and treatment of stroke is a difficult problem in the world. The fundamental purpose of the treatment is to promote cells in the ischemic regional to survive. The study shows that cells can degrade intracellular organelles, abnormal or toxic substances, then provide energy and substrate metabolism to cells to promote cell survival through autophagy. The fusion of autophagosome with lysosome is the most important step in autophagy, only the two were completed fused, the enzymes in the lysosomal can work on the material in the autophagosomes. Autophagosomes must rely on SNAP29 to adhere to lysosome ,so SNAP29 is a key protein in autophagy. Our previous study found that the level of autophagy is higher in the ischemic penumbra after hyperbaric oxygen pretreatment, and there is more autolysosome. So, Is Hyperbaric oxygen preconditioning promoted formation of autolysosome through SNAP29 to be neuroprotective? Therefore, We focus on the formation of autolysosome, using specific fluorescence labeled GFP-LC3 transgenic mice, with the help of electron microscopy and immunohistochemical techniques to explore the specific molecular mechanism of how Hyperbaric oxygen preconditioning promoted autophagy ,and then provide a theoretical basis for hyperbaric oxygen preconditioning on the treatment of stroke.

中风的预防及治疗是一项世界难题。中风治疗的根本宗旨是促使缺血区域受损细胞存活。研究表明：自噬可降解细胞内受损细胞器及有毒或异常物质，并向细胞提供能量和代谢底物，促使细胞存活。自噬体与溶酶体融合形成自噬-溶酶体是自噬反应最重要的步骤，只有二者完成融合后，溶酶体内的酶类才能对自噬体内的待降解物质发挥水解作用。自噬体必须借助SNAP29与溶酶体粘附，才能进行融合，因此SNAP29是关乎自噬反应能否正常进行的的关键蛋白。我们前期研究发现高压氧预处理后缺血半暗带区自噬水平升高、融合后的自噬-溶酶体数量增多。那么高压氧预处理是否是通过SNAP29调控自噬-溶酶体融合、调节自噬水平，从而发挥脑保护效应的呢？为此，本项目拟以自噬-溶酶体形成为关注点，应用特异性荧光标记GFP-LC3转基因小鼠，采用免疫荧光、电镜及组化等技术，探索高压氧预处理促进自噬的具体分子机制，为高压氧治疗缺血性脑中风提供理论依据。

中风的预防及治疗是一项世界难题。中风治疗的根本宗旨是促使缺血区域受损细胞存活。研究表明：自噬可降解细胞内受损细胞器及有毒或异常物质，并向细胞提供能量和代谢底物，促使细胞存活。自噬体与溶酶体融合形成自噬-溶酶体是自噬反应最重要的步骤，只有二者完成融合后，溶酶体内的酶类才能对自噬体内的待降解物质发挥水解作用。自噬体必须借助SNAP29与溶酶体粘附，才能进行融合，因此SNAP29是关乎自噬反应能否正常进行的的关键蛋白。本研究结果显示高压氧预处理通过上调自噬水平，发挥神经保护作用；高压氧预处理促进自噬溶酶体融合，上调自噬水平；高压氧预处理通过SNAP29调控自噬体与溶酶体融合，调节自噬水平，发挥神经保护作用。探索高压氧预处理促进自噬的具体分子机制，为高压氧治疗缺血性脑中风提供理论依据。