Lipopolysaccharide (LPS) is a bacterial wall endotoxin producing many pathophysiological conditions including myocardial inflammation leading to cardiotoxicity. Arachidonic acid is a polyunsaturated fatty acid that can be metabolized to cardioprotective epoxyeicosatrienoic acids (EETs) by cytochrome P450 epoxygenases. These metabolites are subsequently hydrolyzed to less bioactive dihydroxyeicosatrienoic acids (DHET) by soluble epoxide hydrolase (sEH). EETs are known to trigger a wide range of pathways protecting cellular structures, reducing cell death and promoting anti-inflammatory reactions in various cell types. We have recently demonstrated that EETs protect rat neonatal cardiomyocytes against LPS-induced cytotoxicity. Increased exposure to LPS as an environmental toxin can have a negative impact on the heart resulting in cardiovascular complications. The goal of this study is to investigate whether inhibition of sEH, genetically or pharmacologically, will influence mitochondrial function following LPS exposure. Age matched 2 month old sEH null (KO) and littermate wild-type (WT) mice were injected with LPS (10mg/kg) then sacrificed after 6 or 24 hrs. Hearts and blood were collected to assess inflammatory response and mitochondrial function. Mitochondrial function was evaluated by measuring the levels of ATP in the myocardium and respiratory activity of isolated cardiac mitochondria using a Clark-type electrode. The levels of glucose and inflammatory cytokines in blood were also assessed. Our data demonstrated that LPS-triggered a massive inflammatory response beginning at 6h and until 24h in WT mice. This coincided with pronounced hypoglycemic response and compromised mitochondrial function in the WT mice. In contrast, sEH-KO mice were protected against LPS-induced cardiotoxicity. These animals did not develop hypoglycemia and no loss in body weight was detected. The levels of inflammatory markers (TNFα, MCP-1) were only modestly elevated and significantly lower than WT mice treated with LPS. Mitochondrial function was preserved in hearts from sEH KO mice based on higher respiratory control ratios compared to WT. Deletion soluble epoxide hydrolase provides protective effects against LPS-induced cardiotoxicity maintaining mitochondrial function. Our data suggest that inhibition of sEH to elevate endogenous levels of EETs is a potential therapeutic approach to limiting LPS-induced cardiotoxicity.