OBJECTIVES: The present study aimed to clarify the effects of Gypenosides on myocardial ischemia-reperfusion injury. Using rat H9c2 cardiomyocytes as the research object, the model of cardiomyocyte hypoxia and reoxygenation was established to observe the protective effects of Gypenosides on myocardial ischemia-reperfusion injury, revealing the key targets and possible mechanisms for Gypenosides to exert myocardial protection. MATERIAL AND METHODS: A model of cardiomyocyte hypoxia and reoxygenation was prepared. The activity of cardiomyocytes was detected by CCK-8 method. The cardiomyocyte injury was evaluated by LDH assay. The cardiomyocyte apoptosis rate was detected by flow cytometry. The mitochondrial membrane potential of cardiomyocytes was detected by JC-1 staining. Western blot was used to detect the expression of MPTP downstream apoptotic pathways and MPTP opening-related regulatory factors. RESULTS: The cell survival rate of each Gypenosides pretreatment group was significantly higher than that of the hypoxia-reoxygenation group, indicating that Gypenosides could inhibit cell apoptosis and the decrease of mitochondrial membrane potential of hypoxia-reoxygenation cells. The expressions Cytochrome C, APAFl, Caspase-9, and Caspase-3 proteins were significantly lower than those of the hypoxia-reoxygenation group, the expression of Bax was significantly lower than those of the hypoxia-reoxygenation group, while the expression of Bcl2 was significantly higher than those of the hypoxia-reoxygenation group. CONCLUSION: Gypenosides can effectively reduce myocardial ischemia-reperfusion injury in rats. By regulating Bax and Bcl2, Gypenosides can inhibit MPTP opening and the activation of downstream apoptotic pathways, thereby reducing myocardial ischemia-reperfusion injury.