Tumor necrosis factor-alpha augments contraction and cytosolic Ca(2+) sensitivity through phospholipase A(2) in bovine tracheal smooth muscle.

To elucidate the effects of tumor necrosis factor-alpha (TNF-alpha) on tracheal smooth muscle contraction, we simultaneously measured isometric tension and intracellular Ca(2+) ([Ca(2+)](i)) in fura 2-loaded muscle strips. Smooth muscle force generation was evaluated in a high potassium (K(+); 20.0-80.0 mM) solution and with acetylcholine (3 nM-10 microM ). TNF-alpha (1-100 ng/ml) did not directly contract muscle strips. The contractile response to acetylcholine was enhanced after application of 10 ng/ml of TNF-alpha for 30 min but not the response of [Ca(2+)](i). The contractile response and the response of [Ca(2+)](i) to a high K(+) solution were not altered after application of TNF-alpha. The [Ca(2+)](i)-tension curve indicated that TNF-alpha enhanced the responsiveness of tracheal smooth muscle through the acetylcholine-mediated Ca(2+) sensitivity of intracellular contractile elements. The augmentation of the acetylcholine concentration-response curves for muscle tension in the presence of TNF-alpha (10 ng/ml) was inhibited in part after application of manoalide, a phospholipase A(2) inhibitor. We conclude that a low concentration of TNF-alpha enhances smooth muscle responsiveness to acetylcholine by agonist-mediated Ca(2+) sensitivity facilitated by phospholipase A(2).