Following dexamethasone (DEX), cardiac energy generation is mainly through utilization of fatty acids (FA), with DEX animals demonstrating an increase in coronary lipoprotein lipase (LPL), an enzyme that hydrolyzes lipoproteins to FA. We examined the mechanisms by which DEX augments cardiac LPL. DEX was injected in rats and hearts removed, or isolated cardiomyocytes incubated with DEX (0-8h), for measurement of LPL activity and Western blotting. Acute DEX induced whole body insulin resistance, likely an outcome of a decrease in insulin signaling in skeletal muscle, but not cardiac tissue. The increase in luminal LPL activity after DEX was preceded by rapid non-genomic alterations which included phosphorylation of AMPK and p38 MAPK, that lead to phosphorylation of Hsp25 and actin cytoskeleton rearrangement, facilitating LPL translocation to the myocyte cell surface. Unlike its effects in vivo, although DEX activated AMPK and p38 MAPK in cardiomyocytes, there was no phosphorylation of Hsp25, nor was there any evidence of F-actin polymerization or an augmentation of LPL activity upto 8h after DEX. Combining DEX with insulin appreciably enhanced cardiomyocyte LPL activity, which closely mirrored a robust elevation in both phosphorylation of Hsp25 and F-actin polymerization. Silencing of p38 MAPK, inhibition of PI3-kinase or pre-incubation with cytochalasin D prevented the increases in LPL activity. Our data suggest that following DEX, it is a novel, rapid non-genomic phosphorylation of stress kinases, which together with insulin, facilitates LPL translocation to the myocyte cell surface.