: Severe ketoacidosis induces heart failure and cardiac arrest, but its mechanism is unknown. Recently, hydroxy -carboxylic acid receptor 2 (HCA 2 ) was found to be a receptor for a ketone body, β -hydroxybutyric acid (BHB), and is coupled with Gi -GTP binding protein. HCA 2 expression was reported in the guinea pig heart. Therefore, using guinea pig cardiac myocytes, we investigated effects of BHB on L -type Ca 2+ current pre -augmented with β -adrenoceptor agonist, isoproterenol under the whole -cell voltage clamp. BHB significantly reduced the Ca 2+ current preaugmented with isoproterenol. The effect of BHB was concentration dependent with IC 50 of 1.1 mM. Nicotinic acid (NA), another ligand for HCA 2 , also exerted an effect on the Ca 2+ current similar to that of BHB. The effects of BHB and NA were reduced by a specific Gi inhibitor, pertussis toxin in the pipette solution. Our results suggest that BHB activates Gi -coupled signal transduction pathway via HCA 2 in guinea pig cardiac myocytes. The HCA 2 -mediated signal transduction may be associated with ketoacidosis -induced cardiac suppression.