Aim To explore whether inhibiting P53 and increasing the expression of glucose transporter 4 (GLUT4) in cardiomyocytes can improve glucose metabolism disorder and reduce apoptosis induced by high glucose (HG) combined with ischemia-hypoxia(IH). Methods The cardiomyocyte HG＋IH model was induced in vitro. The experimental groups were the control group, HG group, IH group, HG＋IH group, HG＋IH＋P53 inhibitor group (HG＋IH＋Pifithrin-α group), HG＋IH＋P53 inhibitor＋GLUT4 inhibitor group (HG＋IH＋Pifithrin-α＋Fasentin group). Cell viability was detected by the CCK-8 method, lactate dehydrogenase (LDH), glycolytic key enzyme activity, and ATP content were detected by the kit, protein expression of P53, GLUT4, Bax/Bcl-2 and Caspase-3 were detected by Western blot, and cardiomyocyte apoptosis was detected by flow cytometry. Results ①In the HG+IH cardiomyocyte model in vitro, compared with the control group, the expression of P53 increased by 75%, the expression of GLUT4 decreased by 16%, the content of ATP decreased by 51%, the cell viability decreased by 45%, the LDH activity increased by 3.6 times, the expression of Caspase-3 and Bax/Bcl-2 increased by 54% and 77% respectively, and the apoptosis rate increased (all P＜0.05). ②After inhibiting the expression of P53 in cardiomyocytes, compared with the HG＋IH group, the expression of GLUT4 in cardiomyocytes increased by 34%, the content of ATP increased by 60%, the cell viability increased by 50%, the LDH activity decreased by 13%, the expression of Caspase-3 and Bax/Bcl-2 decreased by 31% and 53% respectively, and the apoptosis rate decreased in HG＋IH＋Pifithrin-α group (all P＜0.05). ③After inhibiting GLUT4, compared with the HG＋IH＋Pifithrin-α group, the expression of GLUT4 in cardiomyocytes decreased by 22%, the content of ATP decreased by 39%, the cell viability decreased by 25%, the LDH activity increased by 21%, the expression of Caspase-3 and Bax/Bcl-2 increased by 43% and 89% respectively, and the apoptosis rate increased (all P＜0.05). Conclusions In the cardiomyocyte model of high glucose combined with ischemia-hypoxia, inhibiting P53 can increase the expression of GLUT4, improve the glucose metabolism disorder of cardiomyocytes induced by high glucose combined with ischemia-hypoxia, and reduce apoptosis.