Protective Effects of Resveratrol against Chronic Immobilization Stress on Testis

  • Bitgul G
  • Tekmen I
  • Keles D
  • et al.
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Abstract

Objective . The aim of this study was to investigate protective effects of resveratrol, a strong antioxidant, against possible negative effects of chronic immobilization stress on testes of male rats histochemically, immunohistochemically, ultrastructurally, and biochemically. Material and Methods . Male Wistar rats were divided into 4 groups ( n = 7 ). Group I, control group (C), was not exposed to stress. Group II, stress group (S), was exposed to chronic immobilization stress. In Group III, low dose resveratrol + stress group (LRS), rats were given 10 mg/kg/day resveratrol just before the stress application. In Group IV, high dose resveratrol + stress group (HRS), rats were given 20 mg/kg/day resveratrol just before the stress application. For chronic immobilization stress application animals were put in the plastic tubes (6 cm in diameter, 15 cm in length) during 32 days for 6 hours. All animals were sacrificed 18 hours after the last stress application. Results . Histochemical and ultrastructural investigations showed that in stress group there was germ cell deprivation in seminiferous tubules and increase of connective tissue on interstitial area. No significant changes were seen in low and high dose resveratrol groups. After immunohistochemical investigations, TUNEL (+) and Active Caspase-3 (+) cells were increased in seminiferous tubules of stress group compared with those control group, but they were decreased in low and high dose resveratrol groups. According to biochemically results, MDA, GSH, and testosterone levels in stress group showed no significant difference when compared with those of the other groups. Conclusion . The chronic immobilization stress increases oxidative stress and apoptosis and causes histological tissue damages; resveratrol can minimize the histological damage in testes significantly.

Figures

  • Figure 1: (a) C group. Normalmorphology of seminiferous tubules (right red arrow) and inside lining smooth seminiferous epithelium (right green arrow) (hematoxylin-eosin ×20). (b) S group. In the seminiferous tubules there are separations between germ cells and the basement membrane (right green arrow) and gaps (right red arrow), and because of pouring degenerated germ cells debris is observed in the lumen of tubules (right black arrow) (hematoxylin-eosin ×20). (c) LRS group. Damages seen in stress group are greatly reduced (right red arrow) and in the lumen of seminiferous tubules mature spermiums are observed. (Hematoxylin-eosin ×20). (d) HRS group. Morphology of the seminiferous tubules is similar to the control group (right red arrow). (Hematoxylin-Eosin ×20).
  • Figure 2: (a) C group. The basal membrane of the seminiferous tubule (right red arrow) has normal thickness (PAS ×100). (b) S group. In interstitial area connective tissue is increased (red bordeed star); diameter of the seminiferous tubule is reduced and basement membrane of seminiferous tubule (right red arrow) has become thicker than that of S group (PAS×100). (c) LRS group. Basementmembrane thickness (right green arrow) and diameter of seminiferous tubules are similar in the C group (PAS ×100). (d) HRS group. Basement membrane thickness (right green arrow) and diameter of seminiferous tubules are similar in the C group (PAS ×100).
  • Figure 3: (a) C group. The normal structure of the tunica albuginea (right green arrow) is observed (Masson’s Trichrome ×20). (b) S group. The increase in connective tissue of the tunica albuginea (right green arrow) is observed (Masson’s Trichrome ×20). (c) LRS group. The normal structure of connective tissue is observed in interstitial area (right yellow arrow) and tunica albuginea (right green arrow) (Masson’s Trichrome ×20). (d) HRS group. The normal structure of connective tissue is observed in interstitial area (right yellow arrow) and tunica albuginea (right green arrow) (Masson’s Trichrome ×20).
  • Figure 4: (a) C group. TUNEL (+) cells (right yellow arrow) are observed (TUNEL ×20). (b) S group TUNEL (+) cells (right yellow arrow) are observed. (TUNEL ×20). (c) LRS group. TUNEL (+) cells (right yellow arrow) are observed (TUNEL ×20). (d) HRS group. TUNEL (+) cells (right yellow arrow) are observed (TUNEL ×20).
  • Figure 5: (a) C group. Active caspase-3 (+) cells are (right pink arrow) observed (Active Caspase-3 ×40). (b) S group. Active caspase-3 (+) cells are (right pink arrow) observed (Active Caspase-3 ×40). (c) LRS group. Active caspase-3 (+) cells are (right pink arrow) observed (Active Caspase-3 ×40). (d) HRS group. Active caspase-3 (+) cells are (right pink arrow) observed (Active Caspase-3 ×40).
  • Figure 6: (a) C group. Spermatogonium (stg) and sertoli cells (se) sitting on properly structured basementmembrane (BM) and above primer spermatoyte (st1) are observed (Uranyl acetate-lead citrate ×1250). (b) S group. The basement membrane (BM) is thicker and the gaps (red bordeed star) between germ cells are observed (Uranyl acetate-lead citrate×1250) (c) LRS group.The seminiferous tubule epithelium is similar to the control group (Uranyl acetate-lead citrate ×1250). (d) HRS group. The seminiferous tubule epithelium is similar to the control group and cell-cell connection (right red borded arrow) is observed. (Uranyl acetate-lead citrate ×1250).
  • Figure 7: (a) Changes in rat weights. #: significant differences versus control. ∗, ∗∗, ∗ ∗ ∗: significant differences versus first weights. (b) Johnson’s testicular biopsy score. ∗: significant decrease versus control. ∗∗: significant increase versus stress. (c) Changes in diameter of seminiferous tubules. ∗: significant decrease versus control. ∗∗: significant increase versus stress. (d) Changes in thickness of basement membrane. ∗: significant increase versus control. (e) Number of TUNEL (+) cells. ∗: significant increase versus control. ∗∗: significant decrease versus stress. (f) Number of Active Caspase-3 (+) cells. ∗: significant increase versus control. ∗∗: significant decrease versus stress.

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APA

Bitgul, G., Tekmen, I., Keles, D., & Oktay, G. (2013). Protective Effects of Resveratrol against Chronic Immobilization Stress on Testis. ISRN Urology, 2013, 1–10. https://doi.org/10.1155/2013/278720

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