AbstractBACKGROUNDRapeseed bee pollen has been recognized as a critical treatment for chronic non‐bacterial prostatitis (CNP) and it also can modulate gut microbiota and improve gut health. This study aimed to explore the anti‐prostatitis effects of rapeseed bee pollen with or without wall‐disruption, and to investigate the connection between this treatment and gut microbiota.RESULTSThe results reveal that rapeseed bee pollen can effectively alleviate chronic non‐bacteria prostatitis by selectively regulating gut microbiota, with higher doses and wall‐disrupted pollen showing greater efficacy. Treatment with a high dose of wall‐disrupted rapeseed bee pollen (WDH, 1.26 g kg⁻¹ body weight) reduced prostate wet weight and prostate index by approximately 32% and 36%, respectively, nearly the levels observed in the control group. Wall‐disrupted rapeseed bee pollen treatment also reduced significantly (p < 0.05) the expression of proinflammatory cytokines (IL‐6, IL‐8, IL‐1β, and TNF‐α), as confirmed by immunofluorescence with laser scanning confocal microscope. Our results show that rapeseed bee pollen can inhibit pathogenic bacteria and enhance probiotics, particularly in the Firmicutes‐to‐Bacteroidetes (F/B) ratio and the abundance of Prevotella (genus).CONCLUSIONThis is the first study to investigate the alleviation of CNP with rapeseed bee pollen through gut microbiota. These results seem to provide better understanding for the development of rapeseed bee pollen as a complementary medicine. © 2023 Society of Chemical Industry.