Melatonin Ameliorates 2,4-Dichlorophenoxyacetic Acid Induced Testicular Steroidogenesis Upset in Mice: An <i>In Vivo</i> and <i>In Silico</i> Study

Abstract

2,4-Dichlorophenoxyacetic acid (2,4-D) is used as a selective herbicide and associated with a variety of toxicities in mammals. In contrast, melatonin is an antioxidant that promotes the elimination of free radicals. In the present study, the protective effects of melatonin (10 mg/kg body weight) against 2,4- D (low, mid, and high dose-16.5, 33.0, and 66.0 mg/kg body weight) induced testicular steroidogenesis alteration were examined using in vivo and in silico models. Doses of 2,4-D and melatonin were administered orally for 28 days. The evaluated parameters were body weight, total protein, markers for male reproductive function, and steroidogenesis i.e. testis weight, total lipid, cholesterol, testosterone, 3 beta-hydroxysteroid dehydrogenase, 17 betahydroxysteroid dehydrogenase, total sperm count, sperm motility, and sperm viability along with the histopathology of the testis. The statistical significant value was considered at p&lt;0.05. Molecular docking study was performed for interaction of 2,4-D and melatonin with steroid binding proteins. In vivo results revealed that 2,4-D treatment showed a significant dose-dependent alteration in above all studied parameters. No significant auto-recovery was observed in the withdrawal study, on the contrarily, the altered parameters were normalized and comparable to control when melatonin was given alone and in combination with 2,4-D. In silico results also demonstrated that the binding affinity of melatonin with steroid binding proteins is higher than 2,4-D. Collectively, these in vivo and in silico findings indicated that 2,4-D induced testicular toxicity accompanied by steroidogenesis upset and can be reduced by melatonin significantly by interacting directly and strongly with studied molecular markers.