Protective effect of hesperidin on methotrexate-induced intestinal epithelial damage in rats

Aim: Methotrexate, a folic acid antagonist is widely used for the treatment of a variety of tumors and inflammatory diseases. Methotrexate affects normal tissues which have a high rate of proliferation, including the hematopoietic cells of bone marrow and the gastrointestinal mucosal cells. Reactive oxygen species play an important role in the pathogenesis of methotrexate-induced intestinal damage. Hesperidine is an antioxidant molecule. This study was performed to investigate the effectivity of hesperidin in the prevention of MTX-induced intestinal injury in rats.Materials and Methods: Seventy-eight male Wistar albino rats, 16 weeks old, weighing between 275-420 g, were divided into four groups: (1) rats receiving only saline (n=19), (2) rats receiving only hesperidin (n=19), (3) rats receiving only methotrexate (n=19), and (4) rats receiving methotrexate plus hesperidin treatment (n=21). A single dose of methotrexate (20 mg/kg) was administered to the rats intraperitoneally. In hesperidin treated group, hesperidin was administered by gavage for 5 days. For methotrexate plus hesperidin treated groups, hesperidin was administered by gavage for 5 days after methotrexate treatment. The rats were sacrificed on the 2nd, 4th and 6th day after methotrexate treatment. Tissue samples from the jejunum were taken for histopathological and immunohistochemical analysis.Result: On the 4th day sample reviews showed that villi and cryptas injuries; cellular infiltration; goblet cell depletion and high total intestinal injury score were seen more often compared to the control group (p=0.003, p=0.002, p=0.001, p=0.002, p=0.003). Crypt injury in the methotrexate plus hesperidine group was less than the methotrexate group and was found statistically significant (p=0.002). Staining with inducible nitric oxide synthase in the methotrexate group was significantly increased when compared to the control group (p=0.031). Inducible nitric oxide synthase and interleukin 8 values in the methotrexate plus hesperidine group were less than those in the methotrexate group, and there was significant difference (p=0.019, p=0.036). On the 6th day sample reviews, reepithelisation was seen in methotrexate and methotrexate plus hesperidine groups. Ki-67 proliferation index in the methotrexate group was found less than that in the methotrexate plus hesperidine and control group on 6th day. Myeloperoxidase activity in the methotrexate group was more than other groups, but this was statistically insignificant.Conclusion: Our results confirmed that administration of hesperidin decreased the methotrexate-induced injury to the small intestine. Hesperidin can be used in clinical practice with methotrexate treatment in order to prevent intestinal epithelial injury