Activation of multiple signal transduction pathways by endothelin in cultured human vascular smooth muscle cells

Cellular responses to the vasoconstrictor peptide, endothelin, have been investigated in quiescent cultured human vascular smooth muscle cells (hVSMC). Endothelin caused intracellular alkalinization and activation of the protein synthetic enzyme S6‐kinase, but such responses were not associated with any mitogenic effects of endothelin on hVSMC. In myo ‐[ 3 H]inositol‐prelabelled hVSMC endothelin elicited a rapid increase in inositol bis‐and tris‐phosphates and concomitant hydrolysis of polyphosphoinositol lipids. In [ 3 H]arachidonate‐prelabelled hVSMC endothelin promoted production of diacylglycerol, the early kinetics of which parallelled poly‐phosphoinositol lipid hydrolysis. Such phospholipase C activation by endothelin was sustained in hVSMC with accumulation of inositol polyphosphates being markedly protracted and the decay of diacylglycerol slow. Endothelin promoted extracellular release of [ 3 H]arachidonate‐labelled material from hVSMC which derived via deacylation of both phosphatidylinositol and phosphatidylcholine. This process was inhibited by phospholipase A 2 and lipoxygenase inhibitors, but insensitive to phospholipase C and cyclooxygenase inhibitors. Endothelin‐induced activation of phospholipase C and phospholipase A 2 signal transduction pathways (EC 50 ∼ 5–8 nM for both) in hVSMC apparently proceed in an independent parallel manner rather than a sequential one.