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Articles in PresS, published online ahead of print May 7, 2002
Am J Physiol Heart Circ Physiol, 10.1152/ajpheart.00141.2002
Submitted on February 21, 2002
Accepted on May 6, 2002
1 Neurosurgery, University of Mississippi Medical Center, Jackson, MS, USA
* To whom correspondence should be addressed. E-mail: johnzhang3910{at}yahoo.com.
This study was undertaken to demonstrate the role of RhoA/Rho kinase pathway in ET-1 induced contraction of the rabbit basilar artery. Isometric tension and Western blot were used to examine ET-1 induced contraction and RhoA activation. The upstream effect on ET-1-induced RhoA activity was determined by using ETA and ETB receptor antagonists, and protein kinase C (PKC), tyrosine kinase and PI-3 kinase inhibitors. The downstream effect of ET-1-induced contraction and RhoA activity was studied in the presence of Rho kinase inhibitor Y-27632. The effect of Rho kinase inhibitor on ET-1 induced MLC phosphorylation was investigated by using urea/glycerol/PAGE immunoblotting. We found (1) ET-1 increased RhoA activity (membrane binding RhoA) in a concentration-dependent manner. (2) ETA but not ETB receptor antagonist abolished the effect of ET-1 on RhoA activation. (3) PI-3 kinase inhibitor, but not PKC and tyrosine kinase inhibitors, reduced ET-1-induced RhoA activation. (4) Rho kinase inhibitor Y-27632 (10 µM) inhibited ET-1-induced contraction. (5) ET-1 increased the level of myosin light chain (MLC) phosphorylation. Rho kinase inhibitor Y-27632 reduced the effect of ET-1 on MLC phosphorylation. This study demonstrated that RhoA/Rho kinase activation is involved in ET-1-induced contraction in the rabbit basilar artery. PI-3 kinase and MLC might be the upstream and downstream factors of RhoA activation.
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