真菌病毒AfPV1对寄主黄曲霉抗压力耐受的影响  

Effects of mycovirus AfPV1 on stress tolerance in host Aspergillus flavus

作　　者：江银辉[1] 杨碧 刘翔 田询 禹文峰[1] 齐晓岚[1] 吴昌学[1] JIANG Yinhui;YANG Bi;LIU Xiang;TIAN Xun;YU Wenfeng;QI Xiaolan;WU Changxue(Key Laboratory of Endemic and Ethnic Diseases,Ministry of Education&Key Laboratory of medical Molecular Biology of Guizhou Province,Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学地方病与少数民族疾病教育部重点实验室&贵州省医学分子生物学重点实验室,贵州贵阳550004

出　　处：《贵州医科大学学报》2022年第10期1147-1152,共6页Journal of Guizhou Medical University

基　　金：国家自然科学基金(32000017,31660012);贵州省科学计划项目(黔科合基础-ZK[2021]重点030);贵州省科技厅区域常见疾病与成体干细胞转化研究平台(黔科中引地[2019]4008);贵州省教育厅项目(KY[2021]313)。

摘　　要：目的探讨真菌病毒Af PV1感染后对寄主黄曲霉抗渗透压、氧化、细胞壁压力、紫外照射、巨噬细胞杀灭以及对细胞黏附的影响。方法将脱病毒菌株(LD-F1)及Af PV1感染菌株(LD-F1-b)黄曲霉孢子接种到含有细胞壁胁迫应激试剂、渗透压胁迫试剂和氧化试剂的YGM培养基、30℃黑暗培养5 d、检测病毒Af PV1对黄曲霉抗压力的影响;通过紫外线照射SDA培养基上的黄曲霉孢子,30℃黑暗培养48 h检测病毒Af PV1对黄曲霉抗紫外照射的影响;黄曲霉孢子与巨噬细胞(RAW264.7)和人肺上皮细胞(A549)共培养,比较黄曲霉孢子死亡率和黏附水平;通过黄曲霉孢子与巨噬细胞(RAW264.7)共培养后收集上清液检测白介素-10(IL-10)、IL-6、IL-1β、干扰素-β(IFN-β)、肿瘤坏死因子-α(TNF-α)、一氧化氮(NO)水平。结果真菌病毒Af PV1减弱了黄曲霉对渗透、氧化和紫外线的抵抗压力,但对细胞壁抗压能力无影响;真菌病毒Af PV1的感染能够降低黄曲霉对人肺上皮细胞A549的黏附能力,同时降低黄曲霉抵抗巨噬细胞杀灭能力;黄曲霉感染后,巨噬细胞产生肿瘤坏死因子-α,真菌病毒Af PV1感染黄曲霉会导致巨噬细胞有增加肿瘤坏死因子-α的产生趋势,但结果不显著。结论真菌病毒Af PV1感染能够降低寄主黄曲霉抗压能力。Objective To investigate the effect of mycovirus AfPV1 infection on host Aspergillus flavus(A.flavus)against osmotic pressure,oxidation,cell wall pressure,ultraviolet irradiation,macrophage killing,and cell adhesion.Methods Conidia of virus-free isolate LD-F1 and AfPV1-infected isolate LD-F1-b were inoculated into yeast-glucose medium(YGM)supplemented with a cell wall stress agent,a hyperosmotic stress mediator and an oxidative stress agent,then incubated at 30℃in darkness for 5 days.The effect of virus AfPV1 on the stress resistance of A.flavus was examined.After A.flavus Conidia cultured on SDA medium were irradiated with ultraviolet light and cultured at 30℃in the dark for 48 h,the effect of virus AfPV1 on the resistance of A.flavus to ultraviolet irradiation was measured.Conidia of A.flavus were co-cultured with macrophages(RAW264.7)and human lung epithelial cells(A549),and colonies were counted to compare the conidial mortality and the conidial adherence levels of A.flavus.After co-culturing conidia of A.flavus with macrophages(RAW264.7),the culture supernatant was collected for enzyme-linked immunosorbent assay(ELISA)to detect the IL-10,IL-6,IL-1β,IFN-β,TNF-α,and NO.Results AfPV1 attenuated the resistance of A.flavus to osmotic,oxidative,and UV stresses,but no effected on its resistance to cell wall stress(CR).AfPV1 infection reduced the adherence of A.flavus to human lung epithelial cell line A549 and decreased the ability of A.flavus to resist macrophage-mediated killing.After A.flavus infection,macrophages produced TNF-α.However,infection of A.flavus by the fungal virus AfPV1 resulted in a tendency to increase TNF-αproduction,but the increase was not statistically significant.Conclusion Mycovirus AfPV1 infection can reduce resistance of the host A.flavus to the stresses.

关 键 词：黄曲霉 真菌病毒 压力耐受 巨噬细胞 细胞黏附 细胞因子 

分 类 号：R379.6[医药卫生—病原生物学]