Effects of Different Doses and Time-Dependency of Busulfan on Testes Parameters and Spermatogenesis in a Rat Model: A Quantitative Stereological Study

The current study aimed at evaluating testis parameters and spermatogenesis changes in male rats administrated by different busulfan doses and time to construct a subfertile animal model by stereological methods. In the present study, 150 male Wistar rats randomly divided into 5 groups. All experimental groups were treated by different concentrations of busulfan (0.0, 2.5, 5, 10, and 15 mg/kg). Rats were sacrificed 1, 15, and 30 days after busulfan treatment. The tissue processing was done for stereological study and the results were analyzed by the one-way ANOVA followed by the Duncan test. The most stereological parameters such as testes weight and volume, tubules volume density, interstitial tissue (P<0.05), and germinal epithelium (P<0.01) were significantly reduced by busulfan treatment. Also, at different busulfan doses, the number of spermatogenic cells including spermatogonia (P<0.05), spermatocyte, round and elongated spermatid, and the Sertoli and Leydig cells (P<0.01) significantly decreased, compared with those of the control group. The decline was more obvious in higher busulfan doses and time (from the day 15 to 30) (P<0.05). Most of testicular stereological parameters reduced during 15 days onwards after busulfan treatment in a dose-dependent manner.