Chunlan Sun,Liping Wang,Binjie Zhou,Xialin Jiang,Haiyan Li,Zhangguo Liu
Uncontrolled breeding and improper management of pet cats and dogs have led to the spread of zoonotic Microsporum canis ( M. canis ) infection, which poses challenges to clinical medication. Therefore, there is an urgent need to explore and discover potential fungicides against M. canis . In this study, licochalcone A (LicoA) was prepared by a green and efficient deep eutectic solvent method based on ultrasonic assisted extraction (DES-UAE). The DES-UAE process with choline chloride and propionic acid as solvent was optimized by response surface method with the extraction amount of LicoA as the objective. Following separation and purification, the purity of LicoA reached 99.79 %. Additionally, LicoA demonstrated significant antifungal activity against M. canis with MIC of 4 μg/mL. LicoA inhibited the synthesis of ergosterol in the fungal membrane, destroy the integrity of the cell membrane, lead to the increase of membrane permeability, the release of intracellular electrolytes, the leakage of nucleic acids and proteins, and affect the normal growth of hyphae. ATPase activity was reduced, and mitochondrial membrane potential (MMP) was reduced, resulting in increased levels of ROS, which eventually induced apoptosis in M. canis . Further transcriptome analysis revealed that LicoA treatment affected “cell integrity”, “energy metabolism”, and “oxidative stress” metabolic pathways in M. canis , including cell wall synthesis, ergosterol synthesis, tricarboxylic acid cycle, electron transport chain, and ribosome metabolism, leading to fungal cell apoptosis. These studies establish an efficient method for LicoA extraction and provide a basis for its use as a novel antifungal agent for the treatment of dermatophytosis.
In this work, an eco-friendly ultrasound-assisted deep eutectic solvent (DES) extraction method combined with UPLC-Q-TOF-MS was developed for the efficient enrichment of flavonoids from Begonia fimbristipula . Among 45 screened DESs, betaine-propionic acid (1:1) was selected as the optimal system. The maximum total flavonoid content reached 82.45 mg rutin/g dry weight under optimized conditions (solid-liquid ratio 52 mL/g, 30 min, 57 °C, 240 W). Mechanistic analysis demonstrated that the DES system disrupted plant cell walls and formed strong hydrogen-bond interactions with flavonoids, thereby improving extraction efficiency. Subsequently, UPLC-Q-TOF-MS analysis was performed to profile the phytochemical composition of the purified extract. Ten major phenolic and flavonoid components were identified in the purified flavonoid extract (Bf-FE), clarifying its material basis of bioactivity. In vitro assays showed that Bf-FE exerted potent antifungal activity against Microsporum canis . Mechanistically, Bf-FE destabilized fungal membrane integrity, inhibited ergosterol biosynthesis, impaired mitochondrial function, and induced excessive ROS accumulation, which was further confirmed by transcriptomic analysis. In vivo results validated the antifungal efficacy and biosafety of Bf-FE, which reduced fungal load and modulated epidermal barrier protein expression. This study systematically elucidates the DES-based green extraction mechanism and multi-target molecular antifungal mechanism of B. fimbristipula flavonoids, providing a theoretical basis for developing these flavonoids as natural antifungal lead compounds.
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