Aims Heart failure with preserved ejection fraction (HFpEF) is an increasingly prevalent disease. Physical exercise has been shown to alter disease progression in HFpEF. We examined cardiomyocyte Ca 2+ homeostasis and left ventricular function in a metabolic HFpEF model in sedentary and trained rats following 8 weeks of moderate-intensity continuous training (MICT) or high-intensity interval training (HIIT). Methods and results Left ventricular in vivo function (echocardiography) and cardiomyocyte Ca 2+ transients (CaTs) (Fluo-4, confocal) were compared in ZSF-1 obese (metabolic syndrome, HFpEF) and ZSF-1 lean (control) 21-and 28-week-old rats. At 21 weeks, cardiomyocytes from HFpEF rats showed prolonged Ca 2+ reuptake in cytosolic and nuclear CaTs and impaired Ca 2+ release kinetics in nuclear CaTs. At 28 weeks, HFpEF cardiomyocytes had depressed CaT amplitudes, decreased sarcoplasmic reticulum (SR) Ca 2+ content, increased SR Ca 2+ leak, and elevated diastolic [Ca 2+ ] following increased pacing rate (5 Hz). In trained HFpEF rats (HIIT or MICT), cardiomyocyte SR Ca 2+ leak was significantly reduced. While HIIT had no effects on the CaTs (1-5 Hz), MICT accelerated early Ca 2+ release, reduced the amplitude, and prolonged the CaT without increasing diastolic [Ca 2+ ] or cytosolic Ca 2+ load at basal or increased pacing rate (1-5 Hz). MICT lowered pro-arrhythmogenic Ca 2+ sparks and attenuated Ca 2+ -wave propagation in cardiomyocytes. MICT was associated with increased stroke volume in HFpEF. Conclusions In this metabolic rat model of HFpEF at an advanced stage, Ca 2+ release was impaired under baseline conditions. HIIT and MICT differentially affected Ca 2+ homeostasis with positive effects of MICT on stroke volume, end-diastolic volume, and cellular arrhythmogenicity.