Flow-mediated regulation of endothelin receptors in cocultured vascular smooth muscle cells: an endothelium-dependent effect

Journal of Vascular Research
Eileen M RedmondJ V Sitzmann

Abstract

The objective of this study was to determine the effect of pulsatile flow on endothelin (ET) receptor expression in vascular smooth muscle cells (VSMC) cocultured with endothelial cells (EC). Using a perfused transcapillary coculture system which permits the chronic exposure of cultured EC and VSMC to physiological shear stresses, cocultures were exposed to stepwise increases in flow up to (1) 2 ml/min (low flow: 0.5 dyn/cm2) or (2) 44 ml/min (high flow: 15 dyn/cm2) and maintained for 72 h before SMC and EC were harvested separately. There was a significant increase in [125I]-ET-1 binding in cocultured VSMC exposed to high flow as compared to low flow (Bmax: 75 +/- 22 vs. 152 +/- 10 fmol [125I]ET-1 bound/mg protein) in the absence of any change in the affinity (KD) of ET-1 for its receptor. ET-1 peptide mRNA levels were significantly decreased in EC exposed to high flow. The increase in [125I]ET-1 binding was associated with an increase in ET-A and ET-B receptor mRNA levels and was EC dependent as [125I]ET-1 binding in monocultured VSMC was the same, regardless of flow conditions. However, the amount of [125I]ET-1 binding on VSMC cultured in the absence of EC was significantly greater than that on cocultured VSMC. High flow cau...Continue Reading

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