Protective Role of Hesperidin Nanoparticles and Hesperidin against Methotrexate-Induced Reproductive System Toxicity in Male Albino Rats

Background: In light of the widespread use of the drug methotrexate in the medical field and the lack of awareness of its side effects and its impact on fertility, it was necessary to search for some treatments that do not contain side effects.

Objective: find effective substances from medical plants to reduce the effect of the drug’s side effects on fertility.

Materials and Methods: The study involved a group of 30 adult male rats, rats were divided into five groups: control group These animals were dosed with Normal Saline ,second group animals that were dosed orally with Methotrexate at 1 mg/kg once a week for two consecutive months ,third group dosed orally with hesperidin (62mg/kg). The fourth group were dosed simultaneously with the hesperidin (62mg/kg) and with the 1 mg/kg Methotrexate. The fifth group were dosed with(62mg/kg) hesperidin nanoparticles and simultaneously with 1 mg/kg methotrexate orally for 60 days. The dosage amount was also chosen based on previous research, and an experiment was conducted on hemolysis and antioxidant activity to determine the safety of the dosage concentration.

Results: The Methotrexate caused a decrease in the level of sex hormones and had a negative effect on the level of sperm count and morphology, while hesperidin had a clear effect in raising the level of LH, FSH and testosterone and also sperm count and reducing abnormalities in the sperm, while on the histological level it was found that Methotrexate displays a distorted structure of the testicular tissue. This is characterized by an enlargement of the spaces between the seminiferous tubules, resulting in increased diameters. Additionally, there is a decrease in the size of the epithelial cell layer, cell necrosis, and a scarcity or absence of sperm. The cohort administered with the active compound hesperidin had a typical configuration of testicular tissue, characterized by the presence of tubules. Sperm has an oval morphology and exhibit a consistent and organized arrangement. The tubules are enveloped by a basement membrane that is composed of sperm-generating cells, including dispersed Sertoli cells with triangular nuclei. In close proximity to the cavity, the cells exhibited a larger size and had black, round nuclei, accompanied by spermatids that displayed a round or rectangular form. The presence of sperm was discovered within the interior spaces of the seminal tubules.

Conclusion: The hesperidin nanoparticles had the best effect on increasing fertility, Hesperidin played a role in reducing the amount of damage caused by the drug Methotrexate on the tissues of the reproductive system. Hesperidin had a role in restoring the level of reproductive hormones to a level similar to normal, while the nanoparticles of hesperidin had a greater role in maintaining the level of hormones in a way similar to the control groups and in protecting the reproductive tissues from the damage caused by methotrexate.