Endothelin-1 induced sustained contractions of tracheal smooth muscle involve an activation of protein kinase C.

ABSTRACT

Endothelin-1, a potent vasoconstrictor peptide produces concentration dependent contractions in lamb tracheal smooth muscle. These contractions are not inhibited by low doses (up to 20 μM) of trifluoroperazine and W-7, the calmodulin/myosin light chain kinase (MLCK) inhibitors. At higher concentrations (200 μM), a delayed and poor reversal of isometric tensions results. These relaxations are coupled with a partial dephosphorylation of regulatory myosin light chain (MLC). Preincubation of fiber strips in MLCK inhibitors (200 μM) results in a delayed and attenuated contractile response but without a dephosphorylation of MLC. H-7, a putative protein kinase C antagonist (25–100 μM) abolishes endothelin-1 induced contractile effects rapidly (50% relaxation within 1–3 min). Moreover, such relaxations are accompanied by complete dephosphorylation of MLC. Phorbol 12, 13-dibutyrate, an exogenous activator of protein kinase C potentiates the endothelin induced contractions. Inactive phorbol ester, 4α-phorbol ester does not elicit any contractile response in the muscle. The down regulation of protein kinase C, on the other hand suppresses such potentiated contractile responses. These results suggest that endothelin-1 induced contractile tensions in tracheal smooth muscle are mediated by a mechanism that involves an activation of enzyme protein kinase C.