【摘要】 目的探究不同生长阶段的烟曲霉对果蝇自噬水平的影响。方法用沙保弱平板培养烟曲霉,收获静息孢子;用沙保弱液体培养基震荡培养烟曲霉,在不同时间收获膨胀孢子、菌丝。以3种形态的烟曲霉分组处理转基因果蝇,组织切片观察烟曲霉形态,Rt-PCR(逆转录PCR)检测自噬相关蛋白LC3BⅡ、Beclin-1mRNA的转录水平,Western blot检测LC3BⅡ、Beclin-1的表达量。结果组织切片观察菌的形态与注入时的形态相符,Rt-PCR检测LC3BⅡ、Beclin-1mRNA的转录水平升高,同时,感染膨胀烟曲霉孢子及菌丝组的果蝇LC3BⅡ、Beclin-1蛋白的表达水平升高。结论烟曲霉膨胀孢子及菌丝能显著提高果蝇自噬水平,而静息孢子不能引起果蝇自噬应答。更多还原

【Abstract】 Objective To explore the effects of Aspergillus fumigatus at different growing stages on the level of autophagy in Drosophila melanogaster.Methods Aspergillus fumigatus was cultured with Sabouraud plate to harvest resting spores,and with Sabouraud liquid medium by shaking culture to collect swollen spores and hyphae at different times.Transgenic Drosophila melanogaster were treated with Aspergillus fumigatus in three morphologic statuses respectively.Morphologic observation of Aspergillus fumigatus was conducted with tissue sections.The transcription of autophagy-related protein LC3 BII and Beclin-1mRNA were observed with Rt-PCR(reverse transcription PCR),and Western blot was used to detect the expression of LC3 BII and Beclin-1.Results The morphological characteristics of the bacteria were consistent with those at the time of injection.The transcription levels of LC3 BII and Beclin-1mRNA increased,so did the expression levels of LC3 BII and Beclin-1mRNA in Drosophila melanogaster infected by the swollen spores and hyphae of Aspergillus fumigatus.Conclusion The swollen spores and hyphae of Aspergillus fumigatus can significantly improve the autophagy level in Drosophila melanogaster,while resting spores cannot cause autophagic response in Drosophila melanogaster.更多还原