闽楠精油对两种癣菌抑制活性和抑制机理研究

This study was to clarify the inhibition of PWEOs and reveal inhibition mechanism of dermatophytes. Use culture plates containing PWEOs in vitro tested PWEOs inhibition effect of dermatophytes and the effect of PWEOs on hyphal structure was observed by microscopy; The changes of cell membrane permeability and the degree of lipid peroxidation were reflected by measuring cell physiological indexes. ROS and MMP probe detection of intracellular ROS and MMP change; Time-killing kinetic method was used to monitor the physiological changes of the cells. Finally, WGCNA analysis was used to identify and verify the key genes. The main components of PWEOs are monoterpenes and sesquiterpenoids, and had strong antifungal activity, The minimum inhibitory concentration (MIC) against both dermatophytes (Microsporum gypsum and Epidermophyton floccosum) were 1.5‰. Microscopic observation showed that PWEOs destroy the structure of hyphae. The cell membrane permeability test indicated that the cell membrane of dermatophytes was damaged by PWEOs. Cellular malondialdehyde (MDA) content was positively correlated with the concentration of PWEOs, suggesting that lipid peroxidation of dermatophytes was caused by PWEOs. Fluorescence microscopy images showed excessive production of reactive oxygen species (ROS) and disruption of mitochondrial membrane potential (MMP) in dermatophytes after PWEOs treatment. The Time-kill kinetics of M. gypsum showed significant differences in protein extravasation, extracellular conductivity and intracellular MDA content after three hours of treatment with 1/3MIC PWEOs compared with the control. Five hub genes were identified by weighted gene co-expression network analysis (WGCNA), of which Long chain fatty acid CoA ligase 1 (ACSL1) was significantly up-regulated expressed, Meiotically up-regulated 72 (MUG72) and GDP-mannose transporters gene 1 (GMT1) were significantly down-regulated expressed after PWEOs treatment, which affected the growth and reproduction of M. gypsum. PWEOs significantly inhibited the growth of dermatophytes by damaging hyphal structure, increasing membrane permeability, inducing excessive accumulation of ROS leading to lipid peroxidation, destroying mitochondrial membrane potential, up-regulating ACSL1 and down-regulating MUG72 and GMT1. These results suggest that PWEOs can be used as natural antifungal agents for sustainable applications.