青蒿琥酯上调巨噬细胞抗菌性自噬作用的机制研究

项目名称： 青蒿琥酯上调巨噬细胞抗菌性自噬作用的机制研究

项目编号： No.81202566

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 药物学、药理学

项目作者： 刘鑫

作者单位： 中国人民解放军第三军医大学

项目金额： 23万元

中文摘要： 抗菌性自噬是机体应对病原体感染的重要途径，寻找调节药物并阐明作用机制具有重要意义。本课题组前期研究发现青蒿琥酯（AS）可上调小鼠巨噬细胞的抗菌性自噬，但机制不明。结合AS的药理特性（诱导NOS释放和上调Ca2+水平）和自噬发生的分子调控机制（mTOR和Beclin1的活化），本项目提出AS通过活性氧和/或Ca2+途径激活关键分子mTOR和Beclin1，进而上调抗菌性自噬的假设。为证实该假设，首先检测AS作用后巨噬细胞ROS水平和ROS通路重要蛋白表达、Ca2+浓度和Ca2+通路重要蛋白活化以及mTOR以及beclin1的活化，进而改变ROS和Ca2+水平，观察对AS诱导mTOR和beclin1的活化以及杀菌性自噬作用的影响。最终明确ROS途径和Ca2+途径在AS上调巨噬细胞杀菌性自噬功能中的调控机制，为基于机体自身免疫功能的抗菌策略研究提供新的思路。

中文关键词： 青蒿琥酯；抗菌性自噬；巨噬细胞；ROS；钙离子

英文摘要： Antibacterial autophagy is an essential host immune pathway for combating infection. It is important to find regulatory agents of autophagy with elucidated mechanisms. We have found previously that Artesunate (AS) could upregulate antibacterial autophagy in murine macrophages, while the mechanism is unclear. We supposed in this study that AS exert such a function via induction of reactive oxygen species (ROS) and / or increase of intracellular calcium levels, which activate mTOR and Beclin1, two key regulatory molecules in autophagy. We designed the following experiments to testify the hypothesis. First, levels of ROS and its regulatory factors (NOX2), calcium concentrations its regulatory factors and activation of mTOR and beclin in macrophages after AS incubation will be detected. Then we wll adjust the ROS levels the Ca2+ levels separately to observe whether the antibacterial autophagy is affected. The regulatory mechanism of AS induced antibacterial autophagy will be elucidated with results obtained from the above experiments, which may provide useful data for the development of novel anti bacteria stratery based on host immune functions.

英文关键词： Artesunate；antibacterial autophagy；macrophages；ROS；calcium