Apoptosis in testicular tissue of rats after vasectomy: evaluation of eNOS, iNOS immunoreactivities and the effects of ozone therapy

Abstract

Objective: We aimed to investigate the changes in endothelial nitric oxide synthase (eNOS) and inducible nitric oxide synthase (iNOS) expression and apoptotic index in rat testicular tissue, as well as serum and seminal plasma sex hormone levels after vasectomy, and the effect of ozone therapy (OT).

Material and methods: Adult male Wistar rats were used (n=6 per group). Control (G1), sham for 4 weeks (G2) or 6 weeks (G3), orchiectomy at the 4(th) (G4) or 6(th) (G5) week after left vasectomy, orchiectomy at the 4(th) (G6) or 6(th) (G7) week after bilateral vasectomy, orchiectomy after 6 weeks OT following left (G8) or bilateral (G9) vasectomy, orchiectomy after 6 weeks OT (G10).

Results: In the left testes, while there were increases in eNOS and iNOS immunoreactivity and apoptotic indexes in G4 and G5, no changes were observed in contralateral testis. These values increased in G6 and G7, while OT inhibited these parameters in the left testis of G8 and both testes of G9. Sex hormone levels did not show any changes after vasectomy and ozone therapy.

Conclusion: While OT was found to be protective against some parameters mentioned above under stress conditions, it seemed to cause some harmful effects when used in healthy conditions.

Keywords: Apoptosis; eNOS; iNOS; ozone therapy; vasectomy.

Figure 1.

a–i. eNOS and iNOS immunoreactivity and apoptotic changes in testes. eNOS antigen immunopositive reactions in the early and late round spermatids (arrows) and Sertoli cells (arrowheads). ABC indirect immunoperoxidase test, rabbit anti-eNOS antibody, Mayers hematoxylin counterstaining, Bar=180 μm. (a) Control Group 1; (b) Bilateral vasectomy 6 weeks, Group 7; (c) Bilateral vasectomy + ozone therapy Group 9)]. iNOS expressions in the spermatids, spermatogonia (arrows) and in Sertoli cells (arrowheads). Rabbit anti-iNOS antibody, ABC indirect immunoperoxidase test, Mayers hematoxylin counterstaining, Bar=180 μm; (d) Control Group 1; (e) Bilateral vasectomy 6 weeks Group 7; (f) Ozone therapy Group 10. Apoptotic cells in seminiferous tubule epithelia. TUNEL reaction, no counter staining, Bar=180 μm; (g) Control Group1; (h) Bilateral vasectomy 6 weeks Group 7; (i): Ozone therapy Group 10

Figure 2.

Left testis apoptotic index values in study groups (Means)* control vs. G5, G6, G7, G10, **G10 vs. G1, G2, G3