Zinc has a crucial role in maintaining reproductive health and fertility by preserving the structural integrity of sperm, boosting sperm parameters, and upgrading the quantity of reproductive hormones. The main aim of the current study was to evaluate the implications of zinc supplementation on the reproductive hormone and inspect the serum and testicular ions levels to know how zinc intake can affect testosterone levels through the LH-mediated ions pathway. Twenty-one fully-grown Wistar male rats weighing 180–200 g were used in this study. The rats were divided into three groups. The control group received saline orally for 56 days. Male rats in group 2 received 4.9 mg/kg zinc sulfate orally. Group 3 rats received 18.1 mg/kg zinc sulfate. This study showed that Zn treatment reduced animal body weight change without influencing reproductive organ weight. After Zn intake, testosterone, LH, and FSH levels were significantly higher than in the control group. Molecular docking showed a close association between zinc and two important biomolecules, the Steroidogenic Acute Regulatory (StAR) protein and the Hsd17b3 enzyme, which are necessary to the testosterone pathway. Zinc ions increased significantly in testicular and serum samples. Zinc supplements boost sperm count and reduce deformities. Sperm motility and germinal epithelium thickness decreased with high Zn intake. Our data indicate that 4.9 mg/day of zinc is better than 18.1 mg/day, the upper intake limit. Extreme consumption harms reproductive hormones, calcium ions, and sperm motility.