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Introduction: Previous studies have demonstrated the beneficial effects of treadmill exercise (EX) on osteoporosis, and of hyperbaric oxygen (HBO) on osteoblast and osteoclast formation in vitro. We investigated the effects of HBO and the combination of HBO and EX on osteoporosis in ovariectomized rats. Methods: Forty 3-month-old female Sprague-Dawley rats were randomly divided into 5 groups (n = 8): a sham control group (Control); an ovariectomy group; an ovariectomy with treadmill exercise treatment group; an ovariectomy with HBO treatment group; and an ovariectomy with HBO treatment combined with treadmill exercise group. The HBO exposures were 203 kPa, 85-90% O2, 90 min and the exercise regimen was 20 m·min-1, 40 min·day1, 5° slope. Both treatments were administered once daily, five days a week for 12 weeks until the rats were sacrificed. Results: All three treatments (HBO, exercise, and both combined) significantly promoted the expression of the osteoblast‐related gene and oxidative metabolism-related gene (PGC-1α). They also exerted significant inhibitory effects on the osteoclast‐related mRNA expression (RANKL) and bone resorption marker CTX‐I. Additionally, exercise and the combination exercise-HBO treatment increased serum superoxide dysmutase (SOD) and sclerostin expression. No significant between-group difference was observed. Conclusions: Hyperbaric oxygen, exercise, and the combination ameliorated bone microarchitecture deterioration and ovariectomy-induced bone loss in rats, and these inhibitory effects may be associated with the increased SOD and up-regulated PGC-1α.