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NEGATIVE REGULATION OF QUINONE REDUCTASE 2 BY RESVERATROL IN CULTURED VASCULAR SMOOTH MUSCLE CELLS

1 Resveratrol, a polyphenol in red wine, has a cardioprotective effect. Resveratrol‐targeting protein (RTP) has been purified using a resveratrol affinity column (RAC) and has been identified as quinone reductase type 2 (NQO2). We hypothesize that NQO2 is the target protein of resveratrol in vascular smooth muscle cells (VSMC) and that resveratrol inhibits proliferation of VSMC through its action on NQO2. In the present study, we investigated the correlation between NQO2 regulation and cell proliferation in VSMC in response to resveratrol treatment. 2 The RTP was purified using RAC and was detected with a NQO2 polyclonal antibody. The VSMC were incubated with resveratrol (1, 10 and 50 µmol/L) for 24, 48 and 72 h. Cell proliferation was detected by cell counting and bromodeoxyuridine (BrdU) assay. A lentiviral vector incorporating NQO2 short interference (si) RNA of short hairpin design was constructed and transduced into VSMC. Real‐time quantitative polymerase chain reactionwas used to measure NQO2 mRNA levels; NQO2 expression was determined by western blot analysis. 3 Using RAC, we extracted a 26 kDa protein from aortic smooth muscle, which was referred to as RTP‐26. Proliferation of VSMC was inhibited by resveratrol in a concentration‐ and time‐dependent manner. The mRNA and protein expression of NQO2 was also repressed by resveratrol in a concentration‐ and time‐dependent manner. A similar pattern of inhibition was observed for cells treated with resveratrol (25 µmol/L) as for cells transduced with a lentiviral vector containing siRNA sequences against NQO2. 4 Collectively, these data indicate that the suppression of VSMC proliferation mediated by resveratrol correlates with NQO2 downregulation.