Protective effects of pomegranate (Punica granatum) juice on testes against carbon tetrachloride intoxication in rats

Abstract

Background: Pomegranate fruit has been extensively used as a natural medicine in many cultures. The present study was aimed at evaluating the protective effects of pomegranate (Punica granatum) juice against carbon tetrachloride (CCl4)-induced oxidative stress and testes injury in adult Wistar rats.

Methods: Twenty eight Wistar albino male rats were divided equally into 4 groups for the assessment of protective potential of pomegranate juice. Rats of group I (control) received only vehicles and had free access to food and water. Rats of groups II and IV were treated with CCl4 (2 ml/kg bwt) via the intraperitoneal route once a week for ten weeks. The pomegranate juice was supplemented via drinking water 2 weeks before and concurrent with CCl4 treatment to group IV. Group III was supplemented with pomegranate juice for twelve weeks. The protective effects of pomegranate on serum sex hormones, oxidative markers, activities of antioxidant enzymes and histopathology of testes were determined in CCl4-induced reproductive toxicity in rats.

Results: Pomegranate juice showed significant elevation in testosterone, luteinizing hormone (LH) and follicle stimulating hormone (FSH) those depleted by the injection of CCl4. Activity levels of endogenous testesticular antioxidant enzymes; superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione-S-transferase (GST) and glutathione reductase (GR) and glutathione (GSH) contents were increased while lipid peroxidation (LPO) and nitric oxide (NO) were decreased with pomegranate juice. Moreover, degeneration of germ and Leydig cells along with deformities in spermatogenesis induced after CCl4 injections were restored with the treatment of pomegranate juice.

Conclusion: The results clearly demonstrated that pomegranate juice augments the antioxidant defense mechanism against carbon tetrachloride-induced reproductive toxicity and provides evidence that it may have a therapeutic role in free radical mediated diseases.

Figure 1

HPLC-ESI-MS chromatogram of P. granatum juice.

Figure 2

Effect of P. granatum juice on testes weight and relative testes body weight of rats treated with CCl₄. Values are means ± SEM (n = 7). a: significant change at p < 0.05 with respect to the Con group. b: significant change at p < 0.05 with respect to the CCl₄ group.

Figure 3

Effect of P. granatum juice on lipid peroxidation, expressed as malondialdehyde (MDA), and nitric oxide (NO) contents in testes of rats treated with CCl₄. Values are means ± SEM (n = 7). a: significant change at p < 0.05 with respect to the Con group. b: significant change at p < 0.05 with respect to the CCl₄ group.

Figure 4

Effect of P. granatum juice on superoxide dismutase (SOD) and catalase (CAT) activities in testes of rats treated with CCl₄. Values are means ± SEM (n = 7). a: significant change at p < 0.05 with respect to the Con group. b: significant change at p < 0.05 with respect to the CCl₄ group.

Figure 5

Effect of P. granatum juice on serum testosterone, luteinizing hormone and follicle stimulating hormone in rats treated with CCl₄. Values are means ± SEM (n = 7). a: significant change at p < 0.05 with respect to the Con group. b: significant change at p < 0.05 with respect to the CCl₄ group.

Figure 6

Histological changes in the testes of rats. (A) Control testes with normal spermatogenesis. (B) Rats treated with CCl₄ with prominent inflammation, complete swallowing of seminiferous tubules and degenerated germ cells. (C) Rats supplemented with Pom. (D) Rats treated with the Pom and CCl₄. Sections were stained with hematoxylin and eosin (400x).