Matrine Exerts Antifungal and Antibiofilm Effects on Candida albicans via the Oxidative Stress-MAPK-Metabolism Axis

Aim of the study Matrine is a major alkaloid component of the traditional Chinese medicinal plant Sophora flavescens . Although it has been reported to inhibit fungal growth in vitro, its underlying mechanisms of action and potential molecular targets in Candida albicans remain largely unclear. This study aimed to elucidate the antifungal mechanisms of matrine against C. albicans using flow cytometry, transcriptomics, and untargeted metabolomics, thereby providing a theoretical basis for further mechanistic and translational research. Materials and methods The ΔOD values were measured with a microplate reader, and the half-maximal inhibitory concentrations (IC ₅₀ ) values were calculated by nonlinear regression. The inhibitory effects of matrine were evaluated through time-kill assays, Annexin V-FITC/PI staining, DCFH-DA staining for reactive oxygen species (ROS), cell-cycle analysis, transmission electron microscopy (TEM), and fluorescence-activated cell sorting (FACS). RNA-seq and untargeted metabolomics were employed to explore the underlying antifungal mechanisms, and key pathways and differentially expressed genes were further validated by quantitative real-time PCR (qRT-PCR). Results Nonlinear fitting showed IC ₅₀ values of 57.79-535.60 μg/mL (P = 0.0407), indicating a significant concentration-dependent inhibition. Annexin V-FITC/PI assays showed significant differences in apoptosis between the control and matrine-treated groups at concentrations of 128, 256, and 512 μg/mL (all P < 0.05). TEM demonstrated dose-dependent ultrastructural injuries, including cytoplasmic separation, compromised cell-wall architecture, and severe cell-wall disruption and dissolution at concentrations of 512 and 1,024 μg/mL. RNA-seq identified 2,894 differentially expressed genes, including 1,145 upregulated and 1,749 downregulated genes. The upregulation of genes encoding oxidative-stress-related enzymes ( GPX3 , SOD1 , and CTT1 ) and the unfolded protein response-related transcription factor HAC1 , coupled with marked alterations in lipid metabolites, strongly suggested that matrine induced profound oxidative stress. Mechanistically, matrine-induced endomembrane injury and lipotoxicity were linked to the dysregulation of genes involved in cell-wall and membrane biosynthesis, including CHT1 and FKS2 . Conclusion This study demonstrates that matrine exerts potent antifungal effects against C. albicans through an oxidative stress-MAPK-metabolism axis. The proposed mechanism involves the accumulation of ROS, disruption of lipid and redox homeostasis, inhibition of chitin synthase-related cell-wall remodeling ( CHT1 ), and modulation of β-1,3-glucan synthase activity ( FKS2 ). These cascading effects lead to impaired cell-wall integrity, disrupted polarized growth, and suppressed hyphal development, providing a mechanistic basis for further development of matrine as a candidate antifungal agent for candidiasis.