Abstract: Objective To study the possible pathogenesis of infertility caused by di-butyl phthalate (DBP) and investigate the effects of Vitamin C (Vit C) on DBP-induced reproductive function damage in male Sprague Dawley (SD) rats. Methods Sixty male SD rats were divided into 6 groups, including normal control, Vit C (30 mg/kg BW), DBP (100, 500, 1 000 mg/kg BW), and DBP (1 000 mg/kg BW)+Vit C (30 mg/kg BW) groups with ten rats in each group. After the treatment conducted by DBP and Vit C for three weeks, the rats were sacrificed and testicular tissues were collected to calculate organ coefficient and determine sperm motility, concentration, deformity rate. And the index including hematology, and serum chemistry, serum T, LH, FSH concentration were detected. Results Compared with the normal group, the testes of the high-dose DBP group showed obvious atrophy, LDH concentration significantly decreased, the sperm motility and concentration decreased significantly, and the deformity rate of sperm increased, the concentration of T, LH, FSH in serum decreased significantly. Conclusion DBP can cause testicular atrophy in rats, sperm motility, and sperm deformity rate increases. DBP can reduce serum T and LH levels and increase FSH levels. Vit C can improve DBP-induced productive function injury, increase sperm motility and decrease sperm abnormality, elevate the level of serum T, LH, and FSH, and thus promote the reproductive function of the male rat.