The present investigation addressed the impact of the application of olanzapine on the physical and chemical conditions of sperm as a functional unit of the male reproductive system. Data analysis showed a significant variation among the experimental groups in sperm motility, total sperm abnormalities, sperm count by epididymis, and testosterone concentration. Besides, it enlightens the influence of olanzapine on the body weight and relative weight of male reproductive organs such as testis, epididymis, seminal vesicles, and prostate and epididymal fats.
Weight of reproductive organs
The present analysis clarified the significant increase in the relative body weight (290.83 ± 30.13 gm) in olanzapine rats. Also, relative weight of the testis, epididymis, seminal vesicle, and prostate of olanzapine rats expressed significant decline (0.45 ± 0.04 gm), (0.28 ± 0.03 gm), (0.51 ± 0.05 gm), and (0.15 ± 0.02 gm) respectively, in comparison to the control rats. Conversely, the epididymis gland showed more relative adipose tissue (0.97 ± 0.1 gm) than normal epididymis (Table 1).
Table 1
Absolute weight of reproductive organs among the different four rat groups.
Groups | Body | Testis | Epididymal fat | Epididymis | Seminal vesicle | Prostate |
Control | 277.33 ± 20.53 | 1.77 ± 0.04 | 1.89 ± 0.49 | 0.91 ± 0.14 | 1.82 ± 0.09 | 0.66 ± 0.09 |
Olanzapine | 290.83 ± 30.13 | 1.51 ± 0.17 | 2.98 ± 0.62 * | 0.74 ± 0.1 | 1.22 ± 0.31 * | 0.37 ± 0.06 * |
Olanz + ZnSO4 (50mg/kg) | 242.66 ± 25.61 # | 1.56 ± 0.12 | 2.18 ± 0.41 | 0.73 ± 0.08 * | 1.03 ± 0.12 # | 0.38 ± 0.04 # |
Olanz + ZnSO4 (100 mg/kg) | 249.83 ± 22.86 | 1.71 ± 0.22 | 2.04 ± 0.41 # | 0.76 ± 0.01 | 1.17 ± 0.11 # | 0.44 ± 0.04 # |
Zinc sulfate in two doses was found to recover the relative weight of the male reproductive organs to the normal in olanzapine rats. Whereas the seminal vesicle, prostate and body weight were restored (242.66 ± 25.61 gm) by administration of zinc sulfated in olanzapine rats (50µg/kg). Both doses of zinc sulfate signaled non-significant variations in the relative weight of the testis, seminal vesicle, and prostate. While the epididymis of in zinc sulfate-treated rats restored its normal relative weight (0.32 ± 0.03 gm) significantly (Table 2).
Table 2
Relative weight of reproductive organs to the body weight among the different four rat groups.
Groups | Body | Testis | Epididymal fat | Epididymis | Seminal vesicle | Prostate |
Control | 277.33 ± 20.53 | 0.64 ± 0.04 | 0.81 ± 0.06 | 0.39 ± 0.03 | 0.66 ± 0.05 | 0.19 ± 0.01 |
Olanzapine | 290.83 ± 30.13 | 0.45 ± 0.04 * | 0.97 ± 0.1 * | 0.28 ± 0.03 * | 0.51 ± 0.05 * | 0.15 ± 0.02 * |
Olanz + ZnSO4 (50mg/kg) | 242.66 ± 25.61 # | 0.72 ± 0.08 # | 0.82 ± 0.08 # | 0.32 ± 0.03 # | 0.47 ± 0.05 * | 0.18 ± 0.02 # |
Olanz + ZnSO4 (100 mg/kg) | 249.83 ± 22.86 | 0.56 ± 0.05 # | 0.62 ± 0.05 *# | 0.26 ± 0.02 * | 0.46 ± 0.04 * | 0.18 ± 0.02 # |
* Indicates statistically different from the control at p ≤ 0.5, while # indicates statistically different from the Olanzapine group. |
The present investigation addressed the impact of olanzapine on the physical and chemical conditions of sperm as a functional unit of the male reproductive system. Data analysis showed a significant variation among the experimental groups in sperm motility; total sperm abnormalities; sperm count by epididymis, and testosterone concentration.
Here, olanzapine resulted in long oxidative stress with a significant reduction in the vital antioxidant enzymes. Statistics exhibited a significant decline in the levels of antioxidant parameters in olanzapine rats compared to prooxidant levels, which had a considerable increase. Particularly, the SOD activity of olanzapine rats recorded a significant deficiency (40.86 ± 1.89 U/g tissue) in comparison to the control rats. Conversely, the olanzapine rats supplemented with 100µg of zinc sulfate significantly restored the control value (48.80 ± 4.42 U/g tissue) (Fig. 1a).
Catalase significantly decreased in olanzapine rats (12.89 ± 0.48 U/g tissue) compared to the control animals. On the contrary, the higher dose of zinc sulfate of 100µg showed a significant recovery rate of catalase (15.61 ± 0.97U/g tissue) (Fig. 1b).
The vital nonenzymatic glutathione demonstrated a significant decrease in olanzapine rats in comparison to the control one. Besides, the two groups of zinc sulfate significantly improved glutathione concentration in the olanzapine rats (Fig. 1c).
Vitamin C and vitamin E work together to improve the functional quality of sperm, including its motility, counting, and morphology. The concentration of ascorbic acid was significantly reduced (174 ± 13.79µmol/g tissue) in olanzapine rats (Fig. 2a) in comparison with the control animals. Interestingly, either the high or the low zinc sulfate doses significantly restore the ascorbic acid level to the control values. Similarly, vitamin E concentration was significantly decreased by olanzapine as an antischyopatic drug (5.01 ± 0.19 µg/g tissue) in comparison to the control group. On the contrary, the two therapeutic doses of zinc sulfate exhibited a significant recovery (Fig. 2b).
Due to the vital benefits of Coenzyme Q10 for the fertility and sperm motility of males, it was estimated here. Data revealed that olanzapine significantly reduced the level of Coenzyme 10 (175.16 ± 6.43µmol/g tissue) compared to the control. Either the high or the low zinc sulfate doses significantly restore the Coenzyme 10 level to the control values (Fig. 2c).
For sperm vitality and efficiency of sperm motility, the concentration of phosphatidylcholine must be preserved. Here, we found that olanzapine caused a significant deficiency in the phosphatidylcholine concentration (237.83 ± 20.23 µmol/g tissue) compared to the control. Zinc sulfate, either the high or the low dose, was found to significantly restore the control levels of phosphatidylcholine in olanzapine in comparison to control animals (Fig. 2d).
Taurine is a potential factor responsible for promoting spermatogenesis and improving sperm quality in the testis by enhancing testosterone concentration. Therefore, our study explored the deficiency impact of olanzapine on the concentration of taurine (2.02 ± 0.13µg/g tissue) compared to the control. The 50 &100µg doses of zinc sulfate displayed a significant recovery of taurine in olanzapine rats (Fig. 2e).
Notably, olanzapine as an antipsychotic drug caused the elevation of oxidative stress rate. Consequently, the prooxidant levels were significantly increased due to the elevation rate of oxidative stress. Certainly, malondialdehyde (MDA) is considered the end product of cellular lipid peroxidation, which results from increasing the density of free radicals and is regarded as a marker of oxidative stress. Analysis exhibited the highest level of MDA in olanzapine rats in all experimental groups (47.86 ± 3.26n mol/g tissue) (Fig. 3a). On the contrary, the 50 or 100µg of zinc sulfate was found to partially and non-significantly improve the concentration of the MDA in the olanzapine rats in comparison of the control rats. Similarly, the olanzapine rats treated with 50 or 100µg of zinc sulfate revealed a partial significant recovery of the concentration of the reduced glutathione (GSSG) (Figs. 3b) in comparison to the control animals in a dose-dependent manner. Similarly, the olanzapine rats treated with 50 or 100µg of zinc sulfate revealed a partial non-significant recovery of the concentration of the NO in comparison to the control animals (Figs. 3c).
Olanzapine indirectly influenced ion concentrations such as zinc and iron and energy storage indicator (ATP). The current study displayed a significant decrease in zinc, iron, and ATP levels in olanzapine rats. The data analysis indicates a substantial restoration in zinc, iron, and ATP levels between two administrated groups with zinc sulfate (Fig. 4).
Olanzapine showed a significant decrease (77.33 ± 2.46%) in sperm motility in comparison to the normal rats. Data exhibited a non-significance variation in sperm motility between olanzapine administrated 50µg of zinc sulfate in comparison to the nontreated olanzapine rats. Interestingly, 100 µg of zinc sulfate was found to significantly restore the motility in olanzapine rats to the control value (84.16 ± 5.67%) (Fig. 5a).
Regarding Fig. 5b, the total sperm abnormalities displayed a higher significant increase in olanzapine rats (8.33 ± 0.67%) more than the control ones. Although it was not completely recovered, the total sperm abnormalities were significantly improved by either the 50µg or 100 µg of zinc sulfate in comparison to the nontreated olanzapine rats.
Testosterone concentration significantly decreased (2.27 ± 0.47 ng/ml) in olanzapine rats compared with the other experimental groups. The 100 µg of zinc sulfate significantly recovered the testosterone concentration in olanzapine rats (3.34 ± 0.33 ng/ml) to a value close to the control one (Fig. 5c).
Similarly, the sperm count in the olanzapine rats showed a nonsignificance deficiency (18.83 ± 1.47 x106) in comparison to the control rats. Conversely, the olanzapine rats that were exposed to the zinc sulfate of 50µg or 100 µg exhibited a significance (21.83 ± 1.94), (25.83 ± 1.72), respectively, in comparison to the control rats, in a dose-dependent manner (Fig. 5d).
Histopathological results
Examination of H&E stained-sections of control rats' testis revealed parenchyma was formed of rounded seminiferous tubules surrounded by tunica albuginea. Seminiferous tubules were lined by stratified germinal epithelium (spermatogonia, spermatocytes and spermatids) resting on a regular basement membrane. Most of them had narrow lumina. Leydig cells were found in the narrow interstitial (Fig. 1A & B).
Histopathological examination of testis of Olanzapine group showed disturbed architecture of seminiferous tubules, degenerated and exfoliation germinal epithelium of a seminiferous tubule (spermatogonia, spermatocytes and spermatids) with moderately thickened basement membrane. Note wide interstitial tissue with few Leydig cells and vacuoles (Fig. C & D).
The groups treated with Olz + 50mg/Kg ZnSO4 showed moderate improvement of testicular architecture- seminiferous tubules lined with germinal epithelium; spermatogonia primary spermatocytes, spermatids resting on thin basement membrane. Other seminiferous tubule is seen with more or less normal spermatogenic cell lines. Note mild wide interstitial tissue with few Leydig cells and few vacuoles (Fig. 1E & F).
The rats treated with Olz + 100mg/Kg ZnSO4 showed nearly normal testicular architecture- seminiferous tubules lined with germinal epithelium; (spermatogonia spermatocytes, spermatids) resting on thin basement membrane. Most of them had narrow lumina. Clusters of Leydig cells were found in the narrow interstitial (Fig. 1G & H).