【摘要】 目的探究在烟曲霉感染时Dectin-1是否内在化表达并介导了巨噬细胞的自噬活化,初步明确其作用机制。方法采用Western blot法和免疫荧光技术,观察经β-1,3-D-葡聚糖酶消化前后的烟曲霉孢子刺激下,RAW264.7细胞内Dectin-1与LC3Ⅱ的表达与定位,通过DCFH-DA探针检测消化β-葡聚糖对活性氧(ROS)生成的影响。结果烟曲霉孢子刺激后RAW264.7细胞的Dectin-1与LC3Ⅱ表达水平显著升高,同时二者呈斑点状聚集并共定位于烟曲霉孢子表面;消化β-葡聚糖后Dectin-1与LC3Ⅱ表达量降低,荧光斑点消失,并且ROS的生成受到抑制。结论烟曲霉感染时Dectin-1提高自身内在化表达并诱导了巨噬细胞自噬功能的活化。更多还原

【Abstract】 Objective To investigate whether internalized Dectin-1participates in Aspergillus fumigatus induced macrophage autophagy in RAW264.7cells and explore the possible mechanism.Methods The expression and distribution of Dectin-1and LC3 II in RAW264.7cells were detected by using Western blot and immunofluorescence staining upon stimulation with Aspergillus fumigatus conidia(MOI 5∶1)before and after pretreating withβ-1,3-D-glucanase,and DCFH-DA probe was used to analyze the effect ofβ-glucan digestion on Aspergillus fumigatus-triggered ROS production.Results After stimulated with conidia of Aspergillus fumigates,the expression levels of Dectin-1and LC3 II in RAW264.7cells showed significant increases,and they aggregated to phagosomes which contained Aspergillus fumigatus conidia.Afterβ-glucan was digested,the expressions of Dectin-1and LC3 II reduced,and the production of ROS was inhibited.LC3 diffused in cytoplasm.Conclusion Dectin-1increases the expression of internalized Dectin-1and induces macrophage autophagy in Aspergillus fumigatus infection.更多还原