Background: One of the major mechanisms underlying myocardial injury during ischemia and hypoxia is oxidative stress. Under hypoxic conditions, excessive production of reactive oxygen species (ROS) disrupts the cellular antioxidant defense system, enhances lipid peroxidation, and ultimately leads to apoptosis and cellular damage. Superoxide dismutase (SOD) is a key intracellular antioxidant enzyme that neutralizes oxidative stress, whereas malondialdehyde (MDA), an end product of lipid peroxidation, reflects the extent of membrane damage. Therefore, investigating antioxidant compounds in hypoxia-induced cardiomyocyte injury is of significant importance. Pinocembrin, a bioactive compound derived from Dalbergia odorifera T.Chen, exhibits multiple pharmacological activities, including antioxidant and anti-apoptotic effects. Aim: To evaluate the effects of pinocembrin on oxidative stress in hypoxia-induced H9C2 cardiomyocytes. Materials and Methods: H9C2 cardiomyocytes were divided into five groups: a control group, a model group, and three pinocembrin-treated groups (low, medium, and high doses). Except for the control group, cells were cultured under hypoxic conditions (5% CO2, 1% O2, 37 °C) in glucose- and serum-free DMEM to establish a hypoxia-induced injury model. Cells were then treated with pinocembrin for 4 hours. Oxidative stress levels were assessed by measuring ROS accumulation, SOD activity, and MDA content. Result: Compared with the control group, the model group showed a significant increase in ROS production, a decrease in SOD activity, and a marked elevation in MDA levels, indicating severe oxidative stress under hypoxic conditions. In contrast, the pinocembrin-treated groups exhibited attenuation of hypoxia-induced oxidative stress. Specifically, ROS accumulation decreased, SOD activity was restored, and MDA levels were significantly reduced. These findings suggest that pinocembrin exerts protective effects by enhancing the cellular antioxidant defense system and inhibiting lipid peroxidation. Conclusion Pinocembrin reduces oxidative stress and protects against cellular injury in hypoxia-induced H9C2 cardiomyocytes. These results indicate that pinocembrin may serve as a potential therapeutic agent for ischemiaand hypoxia-related cardiac disorders and provide a valuable foundation for further research.