论文 mTOR通路在游离SiO2粉尘诱导肺泡巨噬细胞自噬中的机制研究.doc

学 号：201095110119 毕业论文 GRADUATE THESIS 设计题目:mTOR通路在游离SiO2粉尘诱导肺泡巨噬细胞自噬 中的机制研究 2015年06月04日 摘 要 目的：通过体外培养大鼠肺泡巨噬细胞，使用mTOR抑制剂雷帕霉素抑制mTOR信号通路，游离SiO2粉尘诱导自噬，观察巨噬细胞自噬水平的差异，进一步明确在细胞水平上该信号通路在游离SiO2粉尘诱导肺泡巨噬细胞自噬活动中的机制。 方法：选取SPF级健康雄性Wistar大鼠。麻醉后，采用支气管肺泡灌洗法，离心获得肺泡巨噬细胞，贴壁培养纯化。将肺泡巨噬细胞分为对照组，矽尘组，矽尘+雷帕霉素组。运用Western blotting测定mTOR蛋白LC3蛋白表达水平共聚焦荧光显微镜观察巨噬细胞自噬水平。Objectives:By culturing rat alveolar macrophages in vitro，Using the mTOR inhibitors rapamycin inhibit mTOR signaling pathways，Free SiO2 dust induce alveolar macrophages autophagy. Then observe the differences in the levels of macrophage autophagy, further clear the manifestation of the signaling pathways on the cell level in free SiO2 dust induced alveolar macrophages activity. Methods:Select SPF health male Wistar rats. After anesthesia, using the method of bronchoalveolar lavage, centrifuging to get alveolar macrophages, adherent culture and purification. The alveolar macrophage is divided into control group, silicon dust group, silicon dust + rapamycin group. Using Westem blotting determinate mTOR protein and LC3 protein expression levels of each group. Using confocal fluorescence microscopy?obsive the level of macrophages autophagy. Results: Compared with control group, the mTOR protein expression level of silicon dust and silica dust+rapamycin group decreased obviously, LC3 protein expression level increased significantly. Compared with silicon dust group, the mTOR protein expression of silicon dust+rapamycin group decreased obviously, LC3 protein expression level increased significantly. Conclusions: Free SiO2 dust can induce alveolar macrophages autophagy occurs. When mTOR signaling pathways has been inhibited, the occurrence of autophagy increased significantly, showed that mTOR pathway manifests inhibition effect in the process of free SiO2 dust inducing alveolar macrophages autophagy. Keywords macrophages; autophagy; mTOR signaling pathways 目 录 摘