论文部分内容阅读
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鎓(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。
Objective To investigate whether reactive oxygen species (ROS) participate in the autophagic activation of RAW264.7 cells induced by Candida albicans and elucidate its origin. Methods RAW264.7 cells were cultured in the logarithmic growth phase and treated with 5 kinds of ROS respectively. After the cells were stimulated by Candida albicans, the level of ROS was detected by using dichlorofluorescein diacetate (DCFH-DA) The expression of LC3Ⅱ protein was detected by Western blotting and the expression and localization of LC3 by immunofluorescence. Results After RAW264.7 cells were challenged with Candida albicans, the expression of ROS and LC3Ⅱ in RAW264.7 cells was significantly increased. At the same time, LC3 was spotted and co-localized with Candida albicans. NADPH oxidase (NOX) inhibitor, After treatment with phenyl iodonium (DPI), the expression of ROS and LC3II decreased obviously, and the distribution of LC3 was dispersed in the cells. There was no significant change of ROS levels after other drugs treatment. Conclusion NOX-derived ROS mediates the autophagic activation of RAW264.7 cells under the action of Candida albicans.