Studies have shown that low alcohol is associated with beneficial cardiovascular effects, while high alcohol is proven to be harmful. Previously we have shown that the survival pathway, PI3K, is essential for cardiac inotropy. Therefore, we hypothesized that ethanol may mediate its inotropic effects partially via PI3K pathway. Contractility and [Ca2+]i measurements were performed on adult rat myocytes exposed to low (LA: 5 mM) and high (HA: 100 mM) doses of ethanol (2hr) in the presence or absence of PI3K inhibitor (LY 294002; 1 uM) or PI3K agonist, IGF‐1 (10 uM). LA increased cellular and sarcomeric contraction associated with increase in the velocity of contraction. Relaxation and calcium sequestration was also improved by LA. Inhibition of PI3K negated the contractile effects of LA, but enhanced the relaxation ones. HA decreased the strength of contraction and increased speed of relaxation and calcium sequestration. IGF‐1 reduced the negative contractile effects of HA. On the molecular level, HA increased Akt mRNA expression, however, no changes in protein levels were observed. Our results suggest that the regulation of PI3K activity is crucial for the positive LA‐as well as the negative HA‐induced inotropic effects. Furthermore, the translation of PI3K downstream target, Akt, is decreased with HA, which partly explains the detrimental inotropic effect of alcohol on the heart. Funded by NIH/NIAAA and NIH/NIGMS.