Integrated multi-omics analysis reveals the bioactive components and mechanism of selenium-enriched rapeseed in enhancing male reproductive function

Male infertility is a worldwide health problem, associated with the impediment of spermatogenesis. As a dual-purpose medicinal-edible oil crop, selenium-enriched rapeseed exhibits male reproductive benefits, yet its bioactive components and underlying mechanisms remain unclear. In this study, the extraction procedure was optimized using response surface methodology (RSM). The crude extracts were further fractionated, and water extract (WE) was identified as the most bioactive fraction by bioassay-guided fractionation method. High performance liquid chromatography-inductively coupled plasma-mass spectrometry (HPLC-ICP-MS) analysis revealed methylselenocysteine (MeSeCys) as the predominant selenocompound within WE. Both MeSeCys and WE exhibited spermatogenic enhancement in vitro and in vivo . Mechanistic studies showed that SLC7A11/LRP8 mediated the transport of MeSeCys, while the spermatogenic activity of MeSeCys and WE was related to selenium-glutathione metabolic pathways. However, RNA-sequencing (RNA-seq), qRT-PCR, and WB analyses uncovered WE’s additional regulation of MAPK/ERK signaling, suggesting the presence of other bioactive components. Ultra-performance liquid-chromatography tandem mass spectrometry (UPLC-MS/MS) analysis showed 340 other constituents in WE. Through network pharmacology, computational screening, and cellular experiments, we identified sulforaphane as a potential bioactive compound in WE and demonstrated its synergistic effect with MeSeCys in promoting cell viability. Our findings would contribute to development of selenium-enriched rapeseed with high nutritive value. • Integrated multi-omics analysis revealed active components in Se-enriched rapeseed. • MeSeCys was the main Se species in water extract derived from Se-enriched rapeseed. • Relative to MeSeCys, water extract was also subject to MAPK/ERK signaling pathway. • First evidence for the existence of sulforaphane in WE for spermatogenic enhancement.