活性氧簇在白假丝酵母菌诱导RAW264．7细胞自噬中的作用

目的探究活性氧簇（ROS）是否参与白假丝酵母菌诱导RAW264．7细胞的自噬活化并明确其来源。方法RAW264．7细胞培养至对数生长期并分别以5种ROS生成系统抑制剂处理，白假丝酵母菌刺激细胞后采用二氯荧光素双醋酸盐（DCFH-DA）显示ROS水平，免疫印迹法检测LC3Ⅱ蛋白的表达量，免疫荧光技术观察LC3的表达与定位。结果白假丝酵母菌刺激后RAW264．7细胞的ROS与LC3Ⅱ表达水平显著升高，同时LC3呈斑点状聚集并与白假丝酵母菌共定位；NADPH氧化酶（NOX）抑制剂氯化二亚苯基碘翁（DPI）处理后ROS与LC3Ⅱ表达量明显降低，并且LC3在细胞内弥散分布；其他药物处理后ROS水平无显著变化。结论在白假丝酵母菌作用下NOX来源的ROS介导了RAW264．7细胞的自噬活化。

The role of ROS in Candida albicans induced autophagy of RAW264.7 cells

Objective To investigate whether or not reactive oxygen species （ROS） participates in Candida albicarts induced macrophage autophagy in RAW264.7 cells and to explore the possible source of ROS. Methods RAW264.7 cells in logarithmic phase were treated with 5 kinds of ROS inhibitors and Candida albicans respectively. Intracellular ROS levels were measured with dichiorodihydrofluorescein diacetate （DCFH-DA） dye assay by Fluorescence microscope. The expression of LC311 protein in RAW264.7 cells were detected by Western blot assay. Then the expression and positioning of LC3 were observed by immunofluorescence staining. Results After stimulated with conidia of Candida albicans, RAW264.7 cells showed significant increases of ROS and LC3Ⅱ amount. Interestingly, LC311 was recruited to phagosomes which contained Candida albicans. After the NOX inhibitor DPI blocked the ROS production, the expression of ROS and LC3Ⅱ had no significant increases, and the distribution of LC3 was diffuse in cytoplasm. Conclusion ROS produced by NOX induces RAW264.7 cell autophagy in Candida albicans infection.