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To elucidate the molecular mechanism on anti-arrhythmic function of Dingxin Recipe (DXR). Methods: Cardiac myocytes isolated by trypsin method were cultured and labeled with various fluorescence stains. DXR contained serum induced changes of cellular calcium concentration 〔Ca2+〕i, cell membrane potential (MP) and mitochondria membrane potential (MMP) were determined with laser confocal microscopy. Results: Hypoxia caused increase of 〔Ca2+〕i and MMP, and decrease of MP, while DXR contained serum lowered the 〔Ca2+〕i in normal or hypoxic myocytes, improved hypoxia induced MP reduction and maintain MMP at baseline level under hypoxia circumstances. Conclusion: Through inhibiting the hypoxia caused 〔Ca2+〕i and MMP elevation and MP lowering, DXR displays its action in protecting myocardial cells to prevent and treat arrhythmia.