Effective therapies to reduce ischemia/reperfusion and hypoxia/reoxygenation injury are currently lacking. Borneol has a clear protective effect on ischemia-reperfusion injury, and its protective effect on myocardial hypoxia/reoxygenation and its mechanism have remained to be elucidated. This study investigated the protective effects of borneol on H9C2 cardiomyocytes during hypoxia/reoxygenation injury and to explore the underlying molecular mechanisms. H9C2 rat cardiomyocytes were cultured, the MIRI model was established by hypoxia for 5h and reoxygenation for 24h. The cells were divided into control group, hypoxia/reoxygenation group (H/R group), (low-, medium-, high-) dose borneol group. Borneol administration groups were pre-treated for 12h with different doses of borneol prior to hypoxia/reoxygenation, 24h after reoxygenation, the cells or culture medium were collected for testing. The results showed that borneol has no cytotoxicity and can enhance the viability of cardiomyocytes, significantly increased the SOD content, significantly decreased LDH leakage in cell culture medium, and reduced evidently the apoptosis rate, down-regulated the expression of Bax and Caspase-3. In conclusion, this study demonstrated that borneol has a definite protective effect on hypoxia/reoxygenation-induced injury of rat cardiomyocytes, and its protective mechanism can be related to the inhibition of apoptosis by reducing the expression of Bax and Caspase-3.