We evaluated the relation of segmental and annular tissue Doppler (TD) velocities to serum levels and myocardial gene expression of tumor necrosis factor-α (TNF-α) and inducible nitric oxide (NOS2) in humans. Seven patients with coronary artery disease underwent echocardiographic examination including TD imaging, along with transmural endomyocardial biopsies (2 biopsies per patient for a total of 14 specimens) at the time of bypass surgery. The specimens were analyzed for the number of mRNA copies of TNF-α and NOS2. In addition, serum levels of TNF-α and nitrite were determined before and after surgery. Normal segments (n = 7) had fewer numbers of mRNA copies of both TNF-α (54.6 vs 6.5, p = 0.002) and NOS2 (3,093 ± 486 vs 661 ± 259, p <0.001) than dysfunctional segments (n = 7). Peak systolic velocity (Sm) (13.3 ± 1.4 vs 4.9 ± 1.6 cm/s, p = 0.002) and early diastolic velocity (Em) (16.5 ± 2.7 vs 8.8 ± 1.3 cm/s, p = 0.02) were significantly higher in normal segments. A significant correlation was present among Em, Sm, and the number of mRNA copies of TNF-α and NOS2 at baseline and during infusion of dobutamine (r range -0.72 to -0.92, p <0.01 for all). Likewise, significant relations were present between the changes in serum cytokine levels and changes in the mitral annulus diastolic velocity, E/Ea ratio, and end-diastolic wall stress (r range 0.75 to 0.88, p ≤0.05 for all). In conclusion, Sm and Em are strongly dependent on the regional levels of cytokine gene expression, which are known to have negative inotropic and lusitropic effects when overexpressed.