Antioxidant effects of isorhamnetin 3,7-di-O-beta-D-glucopyranoside isolated from mustard leaf (Brassica juncea) in rats with streptozotocin-induced diabetes

To investigate the effects of isorhamnetin 3,7-di-O-beta-D-glucopyranoside (isorhamnetin diglucoside), a major flavonoid compound of mustard leaf, on oxidative stress due to diabetes mellitus, in vivo and in vitro studies were carried out. Oral administration of isorhamnetin diglucoside (10 or 20 mg/kg of body weight/day for 10 days) to rats with streptozotocin-induced diabetes significantly reduced serum levels of glucose and 5-(hydroxymethyl)furfural (5-HMF), which is glycosylated with hemoglobin and is an indicator of oxidative stress. After intraperitoneal administration, isorhamnetin diglucoside did not show these activities. In addition, after oral administration, the thiobarbituric acid-reactive substance levels of serum, and liver and kidney mitochondria declined significantly compared with the control group in a dose-dependent manner, whereas after intraperitoneal administration these levels fell only slightly. On the basis of the oral and intraperitoneal results, it was hypothesized that isorhamnetin diglucoside was converted to its metabolite in vivo, and its conversion to its aglycone, isorhamnetin, by beta-glucosidase was confirmed; isorhamnetin acted as an antioxidant. Moreover, it was observed that isorhamnetin diglucoside had no effect on the 1,1-diphenyl-2-picrylhydrazyl radical, whereas isorhamnetin showed a potent antioxidant effect in vitro. In addition, intraperitoneal administration of isorhamnetin reduced serum glucose and 5-HMF levels. Furthermore, lipid peroxidation in blood, liver, and kidney associated with diabetes mellitus declined after the administration of isorhamnetin. These results suggest that isorhamnetin diglucoside is metabolized in vivo by intestinal bacteria to isorhamnetin and that isorhamnetin plays an important role as an antioxidant.