The present study examines the effects of apocynin on oxidative stress and antioxidant enzymes in high-fat diet (HFD) induced obese mice. After 12 weeks on HFD, the C57BL/6J mice that clearly exhibited insulin resistance received apocynin (2.4g/L) in their drinking water for five weeks. The results show that apocynin treatment significantly ameliorated hyperglycemia, hyperinsulinemia and dyslipidemia in HFD mice. Furthermore, the intraperitoneal glucose tolerance test (IPGTT) and homeostasis model assessment of insulin resistance (HOMA-IR) indicate significant improvement of insulin sensitivity in HFD mice after apocynin treatment. Compared to the HFD control mice, serum malondialdehyde (MDA) was significantly lower and serum superoxide dismutase (SOD) was significantly higher in apocynin treated HFD mice, indicating that apocynin suppressed systemic oxidative stress in the treated group. In the liver, apocynin significantly reduced the level of MDA. Accordingly, apocynin treatment strengthened the antioxidative defense system with an increased activity of SOD, glutathione-peroxidase (GSHpx) and content of reduced glutathione (GSH). We also found that hepatic catalase (CAT) activity significantly decreased after apocynin treatment which may indicate that apocynin reduces hydrogen peroxide and oxidative stress in the liver. These results suggest that apocynin may ameliorate insulin resistance by reducing systemic and hepatic oxidative stress in HFD fed mice.