Antifertility activity of methanolic bark extract of Aegle marmelos (l.) in male wistar rats

Abstract

Background: Aegle marmelos leaf, seed and fruit from earlier studies is known to affect male fertility in reversible manner. However they had delayed onset and recovery was found to be prolonged. The present study was undertaken with an aim to evaluate the effect of Aegle marmelos bark extract on rats as the extract is found to be a rich source of marmin and fagarine known for reducing male fertility. Three different concentration of methanolic bark extracts of Aegle marmelos (L.) were evaluated for male antifertility activity on albino wistar rats. Methanolic bark extract of Aegle marmelos at the dose of 200, 400, and 600 mg/Kg b.w was administered orally for 60 days. Treatments were stopped thereafter and animals were sacrificed after a recovery period of 30 days. Control animal were administered vehicle (0.5% CMC for 60 days). Lonidamine was used as standard drug to compare the effect of extract.

Results: Methanolic extract causes a dose & duration dependent infertility via reducing reproductive organ weight and serum testosterone levels. Sperm analysis results showed reduction in sperm density, motility, viability and sperm acrosomal integrity without interfering libido and vital organ body weight. Histopathological studies of testes revealed exfoliation of elongated spermatids, nuclear chromatin condensation, degeneration and prominent spaces detected within the germinal epithelium signifying testicular cytotoxicity and necrosis. Time dependent complete infertility was observed in all dose levels. Animals after the withdrawal from treatment, for 30 days showed restoration of the morphological as well as physiological parameters in extract treated rats. Methanolic extract showed lipid lowering activity compared to control, suggestive good candidature of this plant for further studies.

Conclusions: Our studies suggested Aegle marmelos barks methanolic extract as strong candidate for male contraceptive via its ability to produce complete inhibition of pregnancy, rapid restoration of fertility after withdrawal from treatment and its lipid correcting ability proving further beneficial effects.

Figure 1

The Testosterone in rats treated orally with the Methanolic extract of the bark of Aegle marmelos (L.) 200 mg, 400 mg and 600 mg per kg body weight (bw) per day (values are mean ± SEM, n = six animals).

Figure 2

Histology of the testis of a control animal. The seminiferous tubules contain sertoli cells and germ cells of various stages, covering the entire process of spermatogenesis. The lumen contains mature spermatozoa. The interstitium contains distinct leydig cells, scale bar = 10 μm.

Figure 3

Histology of the methanolic extract of the bark of Aegle marmelos (L.) at a dose level 400 mg/kg b.w for 40 days orally treated rat. Shows structural disorganization and mild to modrate exfoliation of elongated spermatids was apparent in some tubules. Large multinucleated cells were also present in a number of tubules, scale bar = 10 μm.

Figure 4

Histology of the methanolic extract of the bark of Aegle marmelos (L.) at a dose level 400 mg/kg b.w for 40 days orally. sertoli cells show vacuolization, whereas the spermatogonia, spermatocytes and spermatids (round as well as elongated) appear normal, with normal nuclear and cytoplasmic characteristics, scale bar = 10 μm.

Figure 5

Histology of the methanolic extract of the bark of Aegle marmelos (L.) at a dose level 400 mg/Kg b.w for 60 days orally. The disruption of spermatogenesis is evident. Vacuolization is evident in the cytoplasm of sertoli cells, spermatocytes and spermatids. The nuclei of spermatocytes and spermatids show pyknosis, scale bar = 10 μm.

Figure 6

Rats withdraw from treated for 30 days after 60 days treatment of methanolic bark extract of Aegle marmelos 400 mg orally with the methanolic extract of the bark of Aegle marmelos (L.) Restoration of spermatogenesis is evident. The seminiferous epithelium is comparable with that of a control animal. The sertoli cells and germ cells show closer association to each other, with normal nuclear and cytoplasmic characteristics. The lumen contains packed spermatozoa, and the leydig cells appear normal, scale bar = 10 μm.