When selectively overexpressed in mouse heart, TNF-α effects the development of a cardiomyopathy that closely mimics that seen in human failing hearts. It has been suggested that two intracellular signaling pathways, the Akt protein kinase and the NF-κB transcription factor, mediated TNF-α signaling. The present experiments assessed the effects of TNF-α overexpression on these two target proteins in vivo. We measured cardiac Akt kinase phosphorylation and NF-κB activity in mice overexpressing TNF-α (TNF1.6). Both basal and insulin-stimulated Akt phosphorylation were reduced by almost 70% by TNF-α overexpression. By contrast, NF-κB was robustly activated. These effects were absent when TNF-α receptor 1 (TNFR1) was selectively ablated. Cardiomyocyte-specific overexpression of the dominant-negative inhibitory κB protein transgene and subsequent inhibition of NF-κB activity attenuated the effects of TNF-α on Akt phosphorylation. NF-κB inhibition also significantly improved fractional shortening and diminished ventricular hypertrophy and survival without affecting infiltrative inflammation or cytokine expression. Thus, while overexpression of TNF-α effected a marked Akt inhibition and NF-κB activation in mouse hearts, inhibition of NF-κB offered salutary benefits mediated at least in part through activation of Akt.