The effects of curcumin on methotrexate-induced ovarian tissue damage in rats

Methotrexate (MTX) is a folic acid antagonist widely used in the treatment of certain cancers and autoimmune diseases. Increased concentrations and chronic use of MTX can lead to inhibition of DNA synthesis and an elevation of intracellular reactive oxygen species (ROS). In our study, it was aimed to investigate the changes in the folliculogenesis process due to oxidative stress in patients using MTX and the effects of curcumin, a powerful antioxidant, in this process. For this purpose, 32 Wistar albino female rats were randomly selected and divided into 4 groups, one control group and three experimental group. MTX was administered i.p. to rats of groups 3 and 4 at a dose of 0.35 mg/kg/day for 28 days. Group 4 subjects were treated with curcumin at 200 mg/kg/day administered by intragastric gavage for 28 days. At the end of the experiment, sections were taken for histopathological analysis of the ovarian tissue obtained from the sacrificed subjects, and Hematoxylin-Eosin (H-E), Periodic acid-Schiff (PAS) and Masson's triple staining were applied to reveal the general characteristics of the ovarian tissue and follicles. In addition, PARP-1 and p53 expression levels as markers of DNA damage and apoptosis were analyzed by immunohistochemical method. The analyzes of malondialdehyde (MDA), which is an oxidative stress marker, and superoxide dismutase (SOD), catalase (CAT), which are major antioxidant enzymes, were performed using commercial ELISA kits. One way Anova and post hoc Tukey tests were used for parametric data in statistical evaluation. Kruskal-Wallis analysis of variance was used for the analysis of non-parametric data. A value of p<0.05 was considered significant for all evaluations. MTX application significantly increased the number of atretic follicles compared to the control group (p=0.011). Curcumin treatment significantly decreased the number of atretic follicles compared to the MTX group (p=0.043). Curcumin treatment significantly changed the increased MDA levels (p=0.031) and decreased SOD values (p=0.005) compared to the MTX group. MTX significantly increased the expression of p53 and PARP-1 compared to group 1 (p=0.017) and group 2 (p=0.002), and curcumin treatment significantly decreased PARP-1 expression compared to the MTX group (p=0.025). The results of our study showed that MTX has negative effects on folliculogenesis through oxidative stress damage mechanisms and DNA synthesis inhibition, and these negative effects can be treated with curcumin, a powerful antioxidant.