维生素D对烟曲霉感染的肺巨噬细胞 自噬机制的初步研究

目的探究维生素D在鼠肺细胞感染烟曲霉后对细胞自噬的影响。方法用一定量的烟曲霉活化孢子感染细胞后,一组细胞加入维生素D(维生素D组),一组细胞加入生理盐水,感染一定时间后用溶酶体探针检测自噬相关分子的表达;收集各组细胞并裂解细胞,离心取上清,用Western-blot法检测上清液中的LC3BII、Dectin-1及ROS的表达水平。结果活化的孢子感染肺巨噬细胞后,维生素D组自噬体与溶酶体共定位减少、吞噬孢子的速率在减少以及ROS水平降低,其对应的胞内Dectin-1、ROS、LC3BII减少且差异有统计学意义。结论烟曲霉感染肺巨噬细胞后,维生素D可通过减弱细胞自噬体与溶酶体的融合并下调自噬信号通路蛋白的表达以达到抵抗烟曲霉感染的作用。

The effect of vitamin D on the autophagy of pulmonary macrophages infected by Aspergillus fumigatus

Abstract: Objective To explore the effect of vitamin D on the regulation of autophagy in pulmonary macrophages infected by Aspergillus fumigatus. Methods After infected with a certain amount of activated Aspergillus fumigatus spores, the pulmonary macrophages were treated with vitamin D (vitamin D group) or with normal saline (control group) respectively for a certain period of time, then the autophagy-related molecules were detected by using lysosome probe. The pulmonary macrophages were collected and lysed; the expression levels of Dectin-1, ROS and LC3B II were detected by using Western-blot. Results After the pulmonary macrophages were infected by activated Aspergillus fumigatus spores, the co-localization of autophagosomes and lysosomes in the vitamin D group decreased, and the corresponding intracellular Dectin-1, ROS and LC3B II reduced with statistical significance. Conclusion After Aspergillus fumigatus infected pulmonary macrophages, vitamin D can inhibit the role of Aspergillus fumigatus by reducing the fusion of autophagosomes and lysosomes and down-regulating the expression of autophagy signaling pathway protein.