Sea buckthorn seed oil triggers mitochondrial dysfunction and apoptosis in <i>Aspergillus niger</i> by activating the AGE-RAGE-like signaling pathway

<h4>Background</h4>Fungal infections, in particular those caused by <i>Aspergillus niger</i> (<i>A. niger</i>), pose a severe threat to human health, and the situation has been getting worse because of increasing resistance against the frontline triazole antifungals. Sea buckthorn (<i>Hippophae rhamnoides</i> L.) seed oil (SBT oil) exhibits potent antifungal activity against <i>A. niger</i>, principally by disrupting mitochondrial function. Its general mode of action is, nevertheless, unclear.<h4>Methods</h4>We employed an integrated approach combining network pharmacology, transcriptomics, molecular docking, molecular dynamics (MD) simulations, and experimental validation to systematically investigate the antifungal mechanism of SBT oil against <i>A. niger</i>.<h4>Results</h4>Network pharmacology identified seven key bioactive compounds and 26 potential targets, with enrichment analysis highlighting the advanced glycation end-products (AGE)-receptor for AGEs (RAGE) signaling pathway as a crucial mechanism. Subsequent transcriptomic and experimental validation in <i>A. niger</i> revealed that SBT oil disrupts a functionally analogous stress response pathway, leading to mitochondrial dysfunction and apoptosis. Transcriptomics revealed 580 differentially expressed genes (DEGs), indicating widespread disruption of metabolic networks. Molecular docking and MD simulations confirmed the stable binding of key compounds to core targets, MAPK3 and BCL2. <i>In vitro</i> experiments showed that SBT oil increases endogenous AGEs, leading to mitochondrial dysfunction, reduced mitochondrial membrane potential (∆Ψm), elevated ROS and MDA levels, inhibited SDH and ATPase activities, and modulation of MAPK3 and METACASP expression within the AGE-RAGE-like pathway in <i>A. niger</i>.<h4>Conclusion</h4>This study provides strong evidence that SBT seed oil exerts its antifungal effects against <i>A. niger</i> primarily through amplification of AGE-RAGE-like signaling, offering a solid foundation for its development as a natural antifungal agent.