Effects and mechanisms of antimicrobial peptide MAF-1A derivatives anti-Candida albicans biofilm in vitro
DENG Si-bo1,2, HUANG Min-hui1,2, ZHANG Ying-chun1, LI Cai-duo1,2, WU Jian-wei1, CHEN Zheng-hong1,2, WANG Tao1,2
1.School of Basic Medical Sciences, Guizhou Medical University, Guiyang 550025,China; 2.Key Laboratory of Medical microbiology and parasitology of Education Department of Guizhou, Guiyang 550025, China
Abstract：The purpose of this study was to investigate the effect and the underlying mechanisms of antimicrobial peptide MAF-1A derivatives against Candida albicans biofilms in vitro. In this research, MIC and MBC of MAF-1A derivatives against C. albicans was determined with microdilution method;the morphological changes of C. albicans biofilm were observed by scanning electron microscopy (SEM) and inverted microscope; the effects of MAF-1A derivatives on biofilm activity at different stages and 80% inhibitory concentration on biofilm (SMIC80) were determined by XTT assay. Flow cytometry, laser confocal microscope and qRT-PCR were used to investigate the mechanism for its anti-C. albicans biofilm activities. The MIC, MFC and SMIC80 of MAF-1A derivatives against C. albicans were lower than those of template peptides, which were 62.5 μg/mL, 125 μg/mL and 62.5~125 μg/mL, respectively. MAF-1A derivatives could significantly inhibit C. albicans adhesion and hyphal development in a dose-dependent manner. The expression of biofilm-related genes (ALS3, HWP1, SUN42, UME6) in C. albicans decreased after treatment with MAF-1A derivatives (tUME6=12.42,P<0.001;tALS3=12.20,P<0.001;tSUN41=7.206,P<0.001;tHWP1=22.52,P<0.001). MAF-1A derivatives can significantly reduce the activity of both the forming and mature biofilms(t250=3.680,P<0.05;t500=4.153,P<0.05;t1000=4.934, P<0.05;t250=0.5335,P<0.05;t500=1.504,P<0.05;t1000=6.431,P<0.05). MAF-1A derivatives with a concentration of 62.5μg/mL could not only directly destroy the mature biofilm structure of C. albicans, but also significantly increase the cell apoptosis and ROS content, and significantly decrease the mitochondrial membrane potential. This study suggests that MAF-1A derivatives have remarkably inhibitory effects on C. albicans biofilm, the mechanisms may be associated with the inhibiting C. albicans adhesion and hyphal development, biofilm damage, promoting cell apoptosis.
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