Glutathione (GSH) is an ubiquitous thiol-containing tripeptide, which plays important roles in cellular protection from oxidative stress. In our search for a dietary source that can increase GSH levels, we discovered that a 24 h treatment of HepG2 cells with rice bran protein hydrolysate (RBPH), prepared by Umamizyme G-catalyzed hydrolysis, increased the GSH content in a dose-dependent manner. RBPH elevated the expression levels of γ-glutamylcysteine synthetase (γ-GCS), which constitutes the rate-limiting enzyme of GSH synthesis, and of another two enzymes, hemeoxygenase-1 (HO-1) and reduced nicotinamide adenine dinucleotide (phosphate): quinone oxidoreductase 1 (NQO1). This induction was preceded by the accumulation of nuclear factor erythroid 2-related factor 2 (Nrf2) inside the nucleus, which is a key transcription factor for the expression of the γ-GCS, HO-1, and NQO1. Pre-treatment of cells with RBPH produced a significant protective effect against cytotoxicity caused by H 2 O 2 or ethanol. These results indicate that RBPH exerts a protective effect against oxidative stress by modulating GSH levels and anti-oxidative enzyme expression via the Nrf2 pathway.Key words rice bran; anti-oxidative; glutathione; nuclear factor erythroid 2-related factor 2 Rice is the most important cereal food in Japan. Rice bran (RB), which constitutes approximately 10% of the grain, is a major by-product of rice milling. RB is rich in protein, lipids, dietary fibers, and vitamins.1,2) Recently, RB was recognized as a functional ingredient, containing such antioxidants as tocopherols, tocotorienols, and γ-oryzanol.3,4) Moreover, enzymatically produced RB protein hydrolysates (RBPHs) were found to possess various biological functions with potential medical applications. In a previous study, we demonstrated that RBPH produced with Umamizyme G, a commercial protease from Aspergillus oryzae, from defatted RB protein exhibited the inhibitory activity of dipeptidylpetidase-IV (DPP-IV) that is a key regulator involved in the prevention and treatment of type 2 diabetes.5) RBPHs produced with other peptidases were shown to have an anti-proliferative effect on cancer cells, 6) and the ability to reduce micellar cholesterol levels.
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