The goal of this research was to reveal the protective effect and mechanism of fibroblast growth factor-1 (FGF1) on oxidative stress injury of H9c2 cells induced by hydrogen peroxide (H₂O₂). The effects of various concentrations of H₂O₂ and FGF1 on the activity of H9c2 cells were analyzed by Real Time Cell Analysis (RTCA). The content of ROS, calcium ion, mitochondrial membrane potential and apoptosis were detected by fluorescence probe, the mRNA expression of Bcl-2, Bax and Caspase-3 were detected by real-time PCR to evaluate whether FGF1 has ability to resist the apoptosis of cardiomyocytes caused by oxidative damage. The results showed that the proliferation of H9c2 cells could be inhibited after being treated with 200 μM H₂O₂ for 12 h, and 100 μg/ml FGF1 could increase the proliferation rate of H9c2 cells, mitochondrial membrane potential and the mRNA expression of Bcl-2, and reduce the ROS accumulation, the level of apoptosis, the content of intracellular calcium and the mRNA expression of Bax and Caspase-3 caused by H₂O₂. The results showed that FGF1 could regulate oxidative stress by improving mitochondrial function and inhibit the H₂O₂-induced apoptosis in H9c2 cells.