Does Crocin Have Beneficial Effects Against Doxorubicin-Induced Testicular Damage in Rats?: Herbal Medicine on Male Fertility

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Poster Session 2
Melike Ozgul Onal (3), Sara Asaad Abdulkareem Aljumaily (1), Gurkan Yigitturk (3), Volkan YASAR (3), Yasemin Bicer (1), Eyup Altinoz (1), Mehmet Demir (2), Hulya Elbe (3), Feral Ozturk (3)
1. Karabuk University, Faculty of Medicine, Department of Biochemistry, Karabuk, TURKEY
2. Karabuk University, Faculty of Medicine, Department of Physiology, Karabuk, TURKEY
3. Mugla Sitki Kocman University, Faculty of Medicine, Department of Histology and Embryology, Mugla, TURKEY

Doxorubicin, crocin, testis, vimentin.

Abstract text

We aimed to evaluate the effects of crocin against doxorubicin-induced testicular damage in rats. MDA (malondialdehyde), GSH (glutathione), SOD (superoxide dismutase), CAT (Catalase), TAS (total antioxidant status) and TOS (total oxidant status) analyses were performed. The measurements of seminiferous tubule diameters were calculated and the testicular damage was evaluated. 

Doxorubicin (DOX) is a wide-spectrum antibiotic used for chemotherapy. The side effects of it have been reported specifically in some tissues such as ovaries, testes and intestinal mucosa. Doxorubicin-induced damage is related with oxidative stress, DNA damage, and apoptosis. These mechanisms effect the spermatogenesis by reducing the testosterone levels, sperm count and sperm motility. Anticancer treatments are crucial for male patients. Crocin is one of the carotenoids that has both anti-inflammatory and antioxidant activities. Crocin is used for the treatments of dysentery, measles, enlargement of the liver and gallbladder, urological infections, asthma and cardiovascular disorders. Vimentin is an intermediate filament, which is found in Sertoli cells, peritubular‐myoid cells and Leydig cells. Its altered distribution in Sertoli cells is associated with impaired spermatogenesis.

40 Wistar albino rats were randomly divided into four groups. Group 1:Control (saline solution 1 ml/kg/24h, i.p. for 15 days), Group 2:Crocin (40 mg/kg/24h, i.p. for 15 days), Group 3:DOX (2 mg/kg/48h i.p. in six injection, cumulative dose 12 mg/kg), and Group 4:DOX+Crocin (DOX 2 mg/kg/48h i.p. in six injection and crocin 40 mg/kg/24h ip for 15 days).  Testis tissues were removed and stained with Hematoxylin-Eosin. The diameters of seminiferous tubules were measured and the damage was evaluated. The mean histopathological damage score (MHDS) was calculated according to the atrophy, edema, vacuolization and disorganization of seminiferous tubules. For this analysis, each slide was semiquantitatively graded as follows: absent (0), mild (1), moderate (2), and severe (3). The maximum damage score was 12. In addition, vimentin expression of Sertoli cells was evaluated by immunohistochemistry and H-Score was calculated. Levels of MDA and GSH, CAT, and SOD activities were determined in tissue. TAS and TOS were calculated. All data are expressed as arithmetic mean±SE. Kruskal-Wallis and Mann-Whitney U tests were used for comparison of data. p<0.05 was regarded as significant. DOX treatment caused significant increases in oxidant status (MDA and TOS) as well as significant decreases in antioxidant systems (GSH, SOD, CAT and TAS) (p<0.05). Administration of crocin with DOX significantly ameliorated all biochemical parameters. In DOX group, we detected atrophy of seminiferous tubules in many areas. The damaged tubules exhibited disorganization, vacuolization, and edema. The MHDSs of control group and DOX group were 0.20±0.13 and 4.60±0.45, respectively. There was a significant difference between these two groups (p<0.05). Crocin administration decreased the histopathologic score. Immunoexpression level of vimentin and seminiferous tubule diameter in DOX+Crocin group significantly increased compared to DOX group, whereas tubule damage significantly decreased (p<0.05, for all). 

Our results reveal that crocin has beneficial effects on DOX-induced testicular damage. Taken together, present study demonstrated that crocin has potential protective effects against DOX-induced testicular damage by modulating oxidant and antioxidant systems, and reorganization of vimentin in Sertoli cells.


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