Effect of rutin and hesperidin on the expression of Nrf2 gene and the activity of hemoxygenase-1 and NAD(P)H-quinone oxidoreductase at their separate and combined action

AbstractFlavonoids rutin (R) and hesperidin (Hes) have a broad spectrum of the biological activity based on their antiradical properties and ability to increase the activity of antioxidant enzymes, including the activity of heme oxygenase-1 (HO-1) and NAD(P)H-quinone oxidoreductase (QR). It is supposed that the main regulator of the activity of HO-1 and QR is the transcription factor Nrf2. The purpose of the study was to determine the effects of R and Hes on the expression of Nrf2 gene and protein, on the activity and mRNA and protein expression of HO-1 and QR at their separate and combined action. Administration in diet of male Wistar rats (with initial body weight 180–200 g) R (400 mg/kg b.w.) and Hes (400 mg/kg b.w.) during 14 days separately or in combination had no toxic or pro-oxidant effect, which were assessed by the level of liver MDA, hydroperoxides of lipids, reduced and oxidized glutathione. R and, to a lesser extent, Hes caused increased activity of HO-1 and QR. Their combined effect on the activity of HO-1 did not differ from the separate effect of each flavonoid. The combined action of R and Hes on the activity of QR was additive. According to Western blotting, changes in HO-1, Nrf2 and QR protein levels under the action of R and Hes separately or in combination were not statistically significant. The results of real time PCR demonstrated the presence of small, but statistically significant, changes in the level of expression of the genes Nrf2 (Nrf2), HO-1 (Hmox1) and QR (NQO1) both for separate and combined action of R and Hes. Thus, the obtained results showed that high-dose of R and Hes separately and in combination didn’t significantly affect the gene and protein expression of transcription factor Nrf2 and that the increased activity of HO-1 and QR was not associated with the increased expression of Hmox1 and NQO1 genes.

Keywords:rutin, hesperidin, heme oxygenase-1, NAD(P)H-quinone oxidoreductase