Evaluating the protective effect of co-administrating of vitamin C and royal jelly on Phenylhydrazine-induced hemolytic anemia's derangements on sperm quality and serum parameters in mice Running Title: Role of vitamin C in anemia

Reduction in blood circulation results in pathological alterations in testicular physiologic function and spermatogenesis. Moreover, free iron ions from RBC lyses and consequently enhancement of iron ions in testicles leads to oxidative stress and affects the functions of reproductive system. In this experimental study, Thirty two mature mice weighting 20-25gr were assigned into 4 groups including control received saline normal (0.2mL, IP), control-sham received phenylhydrazine (PHZ) (6 mg/100g, IP), PHZ+vitamin(Vit) C (250mg/kg, IP)+royal jelly(RJ) (100mg/kg, orally) and group that received RJ+VitC. Following 35 days, the blood samples were obtained from hearth and the serum levels of total antioxidant capacity (TAC) and malondialdehyde (MDA) were evaluated. The sperm parameters including; count, motility, viability, chromatin condensation and DNA damage were assessed as well. Observations demonstrated a significant reduction in TAC level in PHZ gruop, which was remarkably up-regulated by administrating antioxidants. Moreover, the PHZ increased the serum level of MDA. Meanwhile, it was diminished in antioxidants-administrated animals. Finally, co-administrating the Vitamin C and royal jelly resulted in remarkable (P<0.05) enhancement in sperm parameters. In conclusion, our data showed that PHZ by inducing oxidative stress impacted the sperm parameters and co-administrating of Vitamin C and royal jelly significantly up-regulated the serum TAC level and promoted the sperm parameters.