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Vitamin D: An effective way to combat methotrexate-induced testis injury

Alper Yalcin, Hasan Aydin, Ahmet Turk, Mevlut Dogukan, Nadire Eser, Muhittin Onderci, Fatih Uckardes, Atilla Yoldas, Erkan Yilmaz, Hikmet Keles.

Methotrexate (MTX) is a frequently used anticancer drug in the treatment of several diseases. However, MTX therapy causes significant cytotoxicities in testicular tissue. In this experimental study, the therapeutic utility of vitamin D (VD) on MTX induced testicular injury was investigated. Twenty-eight Wistar rats were randomly divided into four equal groups; Control, VD, MTX, and MTX+VD. Following the treatment, the rats were sacrificed and testicular tissues were removed. Testicular tissues were analyzed for routine histopathology and apoptosis. The activities of glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) were evaluated by enzyme-linked immunosorbent assay and malondialdehyde (MDA) levels were measured spectrophotometrically. MTX-treatment group exhibited degeneration of spermatogenic cells, desquamation of the epithelial cells into the lumen of the tubules, lack of spermatozoa in tubule lumen, edema at interstitial area, a statistically significant decrease in both height and diameter of the tubules (p < 0.05). These values were also concordant with the Johnsenís testicular biopsy score (JTBS). In addition, a significantly increased caspase-3 immunoreactivity was observed. Serum and testicular tissue SOD, and GSH-Px enzyme activities were found to be decreased while MDA increased in the MTX group compared to control group (p < 0.05). In the MTX + VD group, the histological injury was reduced, the caspase-3 positivity and MDA level decreased whereas activities of SOD, and GP-x enzymes were significantly increased compared to those in MTX group (p < 0.05). VD supplement may have a therapeutic utility in reducing the MTX-induced cytotoxicities in testicular tissue.

Key words: Methotrexate, testicular injury, vitamin D, caspase-3, oxidative stress

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