The present study was aimed to investigate the effect of hypoxic training on mitochondrial antioxidants and activities of respiratory chain complexes in mitochondria of skeletal muscle in rats. Forty healthy male Wistar rats were randomized to 5 groups (n=8): living low-training low (LoLo), living high-training high (HiHi), living high-training low (HiLo), living low-training high (LoHi), and living high-exercise high-training low (HiHiLo). All the animals were subjected to 5-week training in normoxic (atmospheric pressure=632 mmHg, altitude of about 1 500 m) or hypoxic environment (atmospheric pressure=493 mmHg, simulated altitude of about 3 500 m). Before exhaustive running, the animals stayed in normoxia for 3 d. Skeletal muscles were prepared immediately after exhaustive running. Muscle mitochondria were extracted by differential centrifugation. Spectrophotometric analysis was used to evaluate activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), malondialdehyde (MDA) level and respiratory chain complex (C) I-III activities in muscle homogenate and mitochondria. Results showed that SOD, GSH-Px, CAT activities and MDA level in skeletal muscle homogenate in HiHi and HiHiLo groups were significantly increased (P<0.05 or P<0.01) compared with those in LoLo group. Muscle mitochondrial MDA level in HiHi and HiHiLo groups was significantly lower (P<0.01), while activities of SOD, GSH-Px and CAT were remarkably higher (P<0.01) than those in LoLo group. Meanwhile, C I-III activities in HiHi and HiHiLo groups were increased significantly (P<0.01), and C II activity in HiLo group also was increased remarkably (P<0.01) compared with those in LoLo group. These results suggest that HiHiLo might be an ideal hypoxic training mode.