蜡螟在念珠菌属真菌动物模型中应用价值的探究

The application value of Galleria mellonella in the animal model of Candida

目的构建蜡螟的念珠菌属真菌动物模型并探究真菌对蜡螟的细胞、组织、器官的影响以及念珠菌属真菌蜡螟模型的应用价值。方法用注射器吸取一定量的菌液注入蜡螟体内,在30℃培养箱中培养一定时间后,取出其中的一小部分蜡螟进行解剖,观察真菌菌丝对蜡螟肠道组织的损伤情况,取另一小部分蜡螟做病理切片,同时记录不同时间段蜡螟的生存死亡情况;将空白蜡螟断头取血,分离淋巴细胞后用DMEM培养,用热灭活真菌处理蜡螟淋巴血细胞,37℃培养3 h后,分别用ROS(氧化应激水平)探针和溶酶体探针标记ROS、溶酶体,观察真菌刺激细胞时的自噬水平的变化。结果真菌菌丝对蜡螟的肠道组织产生损伤,病理切片显示大量菌丝生长,细胞吞噬真菌后,ROS升高,溶酶体与菌共定位。结论念珠菌属真菌的蜡螟模型中,真菌对蜡螟的细胞、组织、器官均有损伤,蜡螟的细胞自噬水平升高,并且此模型具有易操作,价格低廉,便于进行统计学分析等优势。

Objective To construct the Galleria mellonella model of Candida species and study the effect of fungi on the cells,tissues and organs of Galleria mellonella,as well as the application value of the model. Methods The Galleria mellonella were injected bacterial suspension and cultivated at,30 ℃ for a certain time. The injury in the intestinal tissue of Galleria mellonellas caused by the fungal hyphae was observed; pathological sections of Galleria mellonellas were made; the mortality and survival of Galleria mellonella at different times were recorded; the lymphocytes of blank control Galleria mellonellas were cultured with DMEM,heat-inactivated and cultured at 37 ℃for 3 hours; ROS（ oxidative stress） probe and lysosomal probe were used to label ROS and lysosomes respectively,and the change in the level of autophagy was observed. Results The intestinal tissue of Galleria mellonella was damaged by the fungal hyphae. Pathological sections showed large amount of fungal hyphae. ROS increased after the phagocytosis,and the co-localization of lysosomes and the Fungi was observed. Conclusion In the Galleria mellonella model of Candida,the fungi damaged the cells,tissue,organs of Galleria mellonella,and the level of autophagy of their cells increased. This model is of low cost and has the advantages of being easy to operate on and convenient for statistical analysis.