1,25-Dihydroxyvitamin D deficiency accelerates male reproductive senescence in aging mice and 1,25(OH)2D3 alleviates oxidative stress via NF-κB/SOD

1,25(OH)₂D₃ has been demonstrated to exert direct actions on male reproductive system in humans or in Animals. With age, renal synthesis of 1,25(OH)₂D₃ declines significantly, and vitamin D supplementation has been found to alleviate the manifestations of male reproductive aging. Therefore, the relationship between 1,25(OH)₂D₃ and male reproductive aging needs further study. To determine whether 1,25(OH)₂D₃ deficiency accelerates male reproductive senescence in aging mice, wild-type and 1α(OH)ase^-/- male mice fed a rescue diet after weaning, and the reproductive phenotypes were evaluated at 12-18 mo of age. We demonstrated that 1,25(OH)₂D₃ deficiency accelerated male reproductive senescence, representing lower fertility efficiency and gonadal hormone levels, reducing cell proliferation, and increasing cell Apoptosis, cellular senescence, and the senescence-associated secretory phenotype (SASP). We confirmed that the increased oxidative stress and DNA damage detected in 1α(OH)ase^-/- mice resulted in accelerated reproductive senescence in reproductive system, since exogenous antioxidant pyrroloquinoline quinone (PQQ) supplementation could largely rescue reproductive aging phenotype. We further validated the antioxidant effect of 1,25(OH)₂D₃ in aging wild-type mice and senescent Leydig cells by treated 18-mo-old wild-type male mice or TM3 cells with 1,25(OH)₂D₃ or vehicle. We assessed the differential gene expression between grouped senescent TM3 cells using RNA-Seq and verified 1,25(OH)₂D₃ exerted an antioxidant role by acting NF-κB/SOD. This study suggests that 1,25(OH)₂D₃ deficiency accelerates male reproductive senescence in aging mice by increasing oxidative stress and 1,25(OH)₂D₃ plays a role in alleviating oxidative stress via NF-κB/SOD signaling pathway.NEW & NOTEWORTHY Based on this studies, we propose that 1,25(OH)₂D₃ can delay male reproductive aging, and we also propose that 1,25(OH)₂D₃ regulates NF-κB to exert antioxidant effect. Therefore, by targeting a fundamental aging mechanism, 1,25(OH)₂D₃ may be an effective agent in maintaining fertility and postponing male reproductive senescence.