新型抗菌肽Mt-22S3对白念珠菌细胞的作用及机制  

Intracellular mechanism of novel antimicrobial peptide Mt-22S3 against Ccandida albicans

作　　者：李彩多 曾晔 石艳萍 张迎春 吴建伟 陈峥宏 王涛 LI Caiduo;ZENG Ye;SHI Yanping;ZHANG Yingchun;WU Jianwei;CHEN Zhenghong;WANG Tao(School of Basic Medicines,Guizhou Medical University,Guiyang 550025,Guizhou,China;Key Laboratory of Pathogen Biology of Institution of Higher Education Institutions of Guizhou Povince,Guiyang 550025,Guizhou,China)

机构地区：[1]贵州医科大学基础医学院,贵州贵阳550025 [2]贵州省普通高等学校病原生物学特色重点实验室,贵州贵阳550025

出　　处：《贵州医科大学学报》2024年第3期375-380,396,共7页Journal of Guizhou Medical University

基　　金：贵州省科技计划项目(黔科合支撑〔2020〕4Y236);贵州省教育厅滚动支持省属高校科研平台项目(黔教技〔2022〕019)。

摘　　要：目的探讨家蝇抗真菌肽-1A(MAF-1A)突变体-22S3(Mt-22S3)对白念珠菌(C.albicans)细胞的作用及机制。方法取对数生长期的C.albicans菌悬液,分别与0、125、250及500 mg/L Mt-22S3作用12 h,采用荧光显微技术及荧光检测法观测Mt-22S3作用C.albicans后细胞内活性氧(ROS)的变化;以荧光检测法测定Mt-22S3作用C.albicans后细胞内线粒体膜电位水平的变化;通过Annexin V-FITC/PI双染法,观察Mt-22S3对C.albicans细胞凋亡的影响;以琼脂糖凝胶电泳阻滞实验分析Mt-22S3对C.albicans细胞DNA的影响。结果经Mt-22S3作用的C.albicans细胞内ROS增高,且随着Mt-22S3浓度升高,细胞内ROS上升(P<0.05);线粒体膜电位检测发现,经Mt-22S3作用的C.albicans线粒体膜电位下降(P<0.01);Mt-22S3可引起C.albicans细胞凋亡及细胞坏死的发生,且Mt-22S3浓度越高,细胞凋亡及细胞坏死现象越明显;凝胶阻滞实验结果显示,Mt-22S3与C.albicans DNA孵育后产生明显的凝胶阻滞现象。结论Mt-22S3可进入C.albicans细胞内发挥抗菌作用,其机制与诱导细胞内ROS积累、去极化线粒体膜、诱导细胞凋亡和死亡、结合DNA等有关。Objective To investigate the intracellular mechanism of Musca domestica antifungal peptide-1A(MAF-1A)mutant-22S3(Mt-22S3)in Candida albicans(C.albicans).Methods C.albicans cells in logarithmic growth were treated with 0,125,250,and 500 mg/L of Mt-22S3 for 12 hours.Fluorescence microscopy and fluorometric assays assessed changes in intracellular reactive oxygen species(ROS)and mitochondrial membrane potential following treatment.The effects on cell apoptosis were determined using the Annexin V-FITC/PI double staining method,while DNA interactions were analyzed via agarose gel electrophoresis.Results Mt-22S3 treatment resulted in a concentration-dependent increase in intracellular ROS(P<0.05)and a decrease in mitochondrial membrane potential(P<0.01),indicating mitochondrial depolarization.Enhanced apoptosis and necrosis were observed at higher concentrations of Mt-22S3.The gel retardation assay revealed significant DNA binding activity by Mt-22S3,suggesting a mode of antimicrobial action.Conclusion Mt-22S3 enters C.albicans cells,inducing ROS accumulation,mitochondrial depolarization,apoptosis,and DNA binding,demonstrating its potential as an effective antimicrobial strategy.

关 键 词：白念珠菌 活性氧 细胞凋亡 抗菌肽 线粒体膜电位 DNA凝胶阻滞 

分 类 号：R978.5[医药卫生—药品]