Tributyl phosphate (TBP) is an emerging hazardous material that is ubiquitous in the environment. However, whether TBP affects the male reproductive system remains unknown. In this study, we exposed male ICR mice and mouse spermatocyte GC-2 cells to TBP to explore the male reproductive toxicity of TBP. The results revealed that TBP (50 and 100 mg·kg·day) decreased sperm count and motility, damaged testicular structure, and induced apoptosis in mice after 35 days of oral gavage. TBP (10, 50, and 200 μM) inhibited cell viability, induced apoptosis, and reduced the ratio of BCL2/BAX in GC-2 cells after 24 h of exposure. TBP (50 and 200 μM) triggered the buildup of reactive oxygen species and induced a reduction in the mitochondrial membrane potential within GC-2 cells. In addition, TBP led to gut microbiota dysbiosis and metabolic profile alterations and increased the abundance of Prevotellaceae, which are negatively associated with sperm motility. Furthermore, TBP caused inflammation, induced the infiltration of Th17 cells into the mouse small intestine, and stimulated IL-17A signaling in mouse testes. In conclusion, TBP induced male reproductive toxicity via the gut-testis axis, oxidative stress, and apoptosis, offering fresh perspectives on TBP's reproductive hazards in male mammals.
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