Pomegranate (P. granatum) fruit juice protects against iron-induced oxidative testicular injury via amelioration of oxidative imbalance and modulation of metabolic indices linked to male infertility

Male infertility is a challenging clinical condition that threatens global reproductive health. Majority of the issues found in testicular dysfunction are associated with oxidative stress. The present study investigated the protective effect of pomegranate fruit extract against Fe²⁺- mediated oxidative testicular injury using ex vivo experimental models. Testicular oxidative injury was induced by incubating testes tissues in Krebs buffer and FeSO₄.7H₂0 solution, and treated by co-incubating with different concentrations (30–240 μg/mL) of pomegranate fruit extract for 90 min (37 °C), with gallic acid serving as the standard antioxidant. Oxidative injury induction led to significant (p < 0.05) elevation of testicular malondialdehyde and LDL-cholesterol concentration, acetylcholinesterase, ATPase, fructose-1,6-bisphosphatase, glycogen phosphorylase and lipase activities. These were concomitantly accompanied by depleted levels of reduced glutathione, glycogen, total cholesterol, triglyceride and HDL-cholesterol and activities of superoxide dismutase, catalase, ENTPDase and 5′Nucleotidase. It also led to distorted testicular histological architecture. Treatment with the fruit extract markedly reversed these levels and activities and preserved testicular morphology. Octadecenoic acid, 1,2,3-propanetriyl ester, (E,E,E)-; squalene and cholesterol, while concomitantly generating 6-Octadecenoic acid, (Z)-; (2Z)-2-Octenoic acid; methyl tetradecanoate; methyl 14-methylpentadecanoate; octadecanoic acid, 2-[(1-oxohexadecyl)oxy]ethyl ester and cholestan-3-ol, 2-methylene-, (3.beta.,5.alpha.)-. This was accompanied by the deactivation of alpha linolenic acid and linoleic acid metabolism, and steroidogenesis. Treatment with pomegranate depleted the oxidative-generated metabolites, while concomitantly generating arachidonic acid, cholesterol margranate and ethyl lithocholate, and reactivating the inactivated pathways. The results indicate the protective effect of pomegranate fruit juice against oxidative testicular injury.