Evaluation of protective effects of resveratrol in rat ovarium against repeated doses of methotrexate

Methotrexate (MTX), a folic acid antagonist, is a chemotherapeutic agent used in the treatment of many diseases in the clinic. Studies have shown that MTX has a cytotoxic effect when used in high doses, and the necessity of preventing the toxicity it causes in organs and tissues has emerged. In our study, we aimed to investigate the protective effects of Resveratrol (RES), which is considered a powerful antioxidant, against MTX-induced ovarian damage in female rats. In our study, a total of 42 female Wistar Albino rats (240-360 g) were used, 7 groups (Group I- Control, Group II- MTX (1st day), GroupIII- MTX (1st and 3rd day), Group IV- MTX (1st, 3rd and 5th day), GroupV- MTX (1st day) + RES, GroupVI- MTX (1st and 3rd day) + RES, GroupVII- MTX (1st, 3rd and 5th day) + RES) have been created. While administering a single dose of MTX 15 mg/kg intraperitoneally to all groups except the control group on the first day; V., VI., and VII. Resveratrol was given to the groups by oral gavage at a dose of 20 mg/kg 1 hour before MTX administration. III., IV., VI. and VII. groups on the 3rd day, IV. and VII. on the 5th day, repeated doses of MTX were administered to the groups. On the 8th day of the experiment, tissue and blood samples were taken from the rats under anesthesia and they were sacrificed. Ovarian damage was evaluated under light microscope by performing hematoxylin-eosin (H-E) staining and TUNEL staining on ovarian tissues of rats. Ovarian tissue samples taken in our study were homogenized and TAS and TOS activities were determined spectrophotometrically. In addition, DNA damage in blood was evaluated by Comet Assay method. Since there was only one animal left in Groups IV and VI and none in Group VII, these groups could not be evaluated. As a result of biochemical analyzes, TOS levels increased in MTX groups, group II and III, compared to the control group, while TOS level decreased significantly in group V. However, the TAS level increased significantly in group V compared to groups II and III. Many findings were improved in group V compared to the MTX groups that were damaged as a result of staining with hematoxylin-eosin. Maximum positive staining in TUNEL image; observed in equal amounts in groups II and III, then in group V. In addition, the comet score showing DNA damage analysis increased significantly in groups II and III, while it decreased significantly in group V. In line with the results of this study, it was observed that resveratrol may provide protective effects against methotrexate-induced ovarian damage due to its antioxidant properties.