摘要
烟曲霉是引起侵袭性曲霉病的关键病原体,严重威胁着人类生命健康。钙调蛋白通路在调控烟曲霉毒力方面起着关键作用,成为抗真菌研究的重要靶点。本研究针对FDA批准的活性化合物库进行系统筛选,成功发现calmidazolium chloride(CMZ)对烟曲霉生长具有显著抑制效果。通过测定不同浓度CMZ对烟曲霉生长、分生孢子存活和萌发、次级代谢以及钙离子信号通路基因表达的影响,全面解析了其抗烟曲霉的作用机制。研究结果表明:CMZ表现出强效的杀菌活性,对烟曲霉的致死率高达(98.96±1.47)%,同时能够抑制(85.26±1.51)%的分生孢子萌发,并导致毒力相关次级代谢产物helvolic acid丰度降低了98.02%。在机制上,本研究揭示了CMZ是烟曲霉钙调蛋白激酶转录抑制剂,通过干扰细胞内钙离子浓度来发挥抗真菌作用。此外,CMZ在与两性霉素B联合使用时,能够发挥明显的协同作用,增强两性霉素B的杀菌作用。进一步研究发现,CMZ对不同来源的病原真菌,包括冻土毛霉、黄曲霉和尖孢镰孢菌均有不同程度的抑制作用,抑制率分别为(24.65±0.90)%、(51.99±0.54)%和(49.98±1.62)%,充分显示出广谱的抗真菌活性。因此,本研究全面评价了钙调蛋白激酶转录抑制剂CMZ的广谱抗真菌活性以及与两性霉素B的协同抗真菌作用,为真菌感染防控提供了极具潜力的候选化合物及联合用药方案。
Aspergillus fumigatus,a leading cause of invasive aspergillosis,poses a severe threat to human health.Calmodulin pathway plays a pivotal role in regulating A.fumigatus virulence,making it a critical target for antifungal research.Here,we systematically screened the FDA-approved compound library and identified calmidazolium chloride(CMZ)as a potent inhibitor of A.fumigatus growth.By assessing the effects of CMZ at varying concentrations on hyphal growth,conidial survival and germination,secondary metabolism,and calcium signaling pathway gene expression,the antifungal mechanisms of CMZ against A.fumigatus were comprehensively elucidated.CMZ exhibited robust fungicidal activity,achieving a mortality rate of(98.96±1.47)%against A.fumigatus,and suppressing(85.26±1.51)%of conidial germination and reducing the abundance of the virulence-associated metabolite helvolic acid by 98.02%.Mechanistically,CMZ functioned as a calmodulin kinase transcription inhibitor,disrupting intracellular calcium concentration to exert antifungal effects.Strikingly,CMZ demonstrated synergistic antifungal activity when combined with amphotericin B,significantly enhancing the fungicidal efficacy of the latter.Furthermore,CMZ displayed broad-spectrum antifungal activity,inhibiting Mucor hiemalis,Aspergillus flavus,and Fusarium oxysporum with suppression rates of(24.65±0.90)%,(51.99±0.54)%and(49.98±1.62)%,respectively.Our study highlights CMZ as a potent calmodulin kinase transcription inhibitor with dual antifungal modes-direct suppression of A.fumigatus and synergistic enhancement of amphotericin B,providing a promising candidate compound and a potential combination therapy strategy for combating fungal infections.
作者
雷雨欣
宋自力
王东
尹文兵
LEI Yuxin;SONG Zili;WANG Dong;YIN Wenbing(Traditional Chinese Medicine College,Shenyang Pharmaceutical University,Shenyang 110016,Liaoning,China;State Key Laboratory of Microbial Diversity and Innovative Utilization,Institute of Microbiology,Chinese Academy of Sciences,Beijing 100101,China;Medical School,University of Chinese Academy of Sciences,Beijing 100049,China)
出处
《菌物学报》
北大核心
2025年第8期116-128,共13页
Mycosystema
基金
国家自然科学基金(32470046)
中国科学院基础研究青年团队项目(YSBR-111)。
