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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 Candida albicans fungi animal model and explore the fungi on the wax beetles cells, tissues and organs, as well as Candida fungal wax borer model of application value. Methods A certain amount of bacterial liquid was injected into the wax borer body with a syringe and cultured in a 30 ° C incubator for a certain period of time. A small part of the wax borer was removed for dissection and the damage of the fungus mycelium to the intestinal tissue of the wax borer was observed. Take a small part of the wax borer for pathological sectioning, and record the survival and mortality of wax borers in different time periods. Take the broken stem of the wax borer to take blood, isolate the lymphocytes and culture them with DMEM. , And cultured at 37 ℃ for 3 h. ROS and lysosomes were labeled with ROS (oxidative stress level) probe and lysosomal probe respectively to observe the change of autophagy level when the fungi stimulated the cells. Results Fungal mycelium damage to the intestinal tissue of wax borers, pathological sections showed a large number of mycelial growth, the phagocytosis of fungi, ROS increased, lysosomes and bacteria co-localization. Conclusion Candida albicans waxy moth model, the fungus on the wax borer cells, tissues, organs are damaged, waxy cell autophagy levels, and this model is easy to operate, inexpensive, easy to carry out statistical analysis And other advantages.