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Background: To investigate the effect of hydrogen peroxide (H 2 S) on myocardial clock gene Bmal1 in ischemic cardiomyocytes., Materials & Methods: Quantitative PCR (qPCR) was used to detect the expression of Bmal1 at the mRNA level in H9C2 rat cardiomyocytes. The protein expressions of Bax and Bcl-2, PI3K/Akt, caspase-3 were measured by western blotting. The levels of reactive oxygen species (ROS) were determined by ELISA., Results: The expression level of clock gene Bmal1 demonstrated a clock rhythm of periodic oscillation within 24 h. Compared with the control group, H 2 S treatment maintained the rhythm of the clock gene in ischemic cardiomyocytes and increased the transcription and expression levels of Bmal1. H 2 S increased cell survival by activating PI3K/Akt signaling pathway, inhibiting mitochondrial apoptosis signaling, and reducing intracellular oxidative stress. PI3K/Akt and Bmal1 were demonstrated to be involved in H 2 S protection of cardiomyocyte ischemia. Knockout of Bmal1 gene affects the degree of phosphorylation of Akt and Erk proteins, and the level of ROS production, resulting in a decrease in the protective effects of H 2 S., Conclusion: The expression level of Bmal1 has effects on the function of cardiomyocytes such as ROS production. The potential mechanism by which H 2 S regulates clock genes may be related to the effect of clock genes on protein phosphorylation levels in ischemic cardiomyocytes., (Copyright © 2019 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)