Abstract
Hypertension is characterized by abnormal vascular contractility and function. Arteries from deoxycorticosterone acetate (DOCA)-salt hypertensive rats develop spontaneous tone that is not observed in arteries from normotensive rats. Inhibition of phosphoinositide 3-kinase (PI3-kinase) by 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002) reduces spontaneous tone development. The Rho/Rho-kinase pathway has been suggested to play a role in hypertension and may be dependent on PI3-kinase activity. We hypothesized that Rhokinase is involved in spontaneous tone development and that Rho/Rho-kinase is a downstream effector of PI3-kinase. Using endothelium-denuded aortic strips in isolated tissue bath, we demonstrated that (+)-(R)-trans-4-(1-aminoethyl)-N-(4-pyridyl) (Y27632) (1 μM), a Rho-kinase inhibitor, significantly reduced spontaneous tone in the DOCA aorta but that it did not affect sham aorta basal tone (DOCA 63.5 ± 15.9 versus sham 1.2 ± 0.4 total change in percentage of phenylephrine contraction). We examined the interaction between the PI3-kinase and Rho pathways by observing the effects of LY294002 on a Rhokinase effector, myosin phosphatase (MYPT), and Y27632 on a PI3-kinase effector, Akt, using Western blot analysis. Inhibition of PI3-kinase reduced spontaneous tone, but it had no effect on the phosphorylation status of MYPT, indicating that PI3-kinase is not a downstream effector of Rho/Rho-kinase. These data indicate that there is little interaction between the Rho/Rhokinase and PI3-kinase pathways in the DOCA-salt aorta, and the two pathways seem to operate in parallel in supporting spontaneous arterial tone. These data reflect spontaneous tone only and do not rule out the possibility of interaction between these pathways in agonist-stimulated tone.
Footnotes
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This study was supported by American Heart Association Established Investigator Award 0240033N to S.W.W. and Midwest Affiliate American Heart Association Predoctoral Grant (1102077) to C.A.N.
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DOI: 10.1124/jpet.103.062265.
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ABBREVIATIONS: PI3-kinase, phosphoinositide 3-kinase; MYPT, myosin binding subunit of myosin phosphatase; phospho-MYPT, phosphorylated form of myosin binding subunit of myosin phosphatase; ET-1, endothelin-1; ROCKI, Rho-kinase I; ROCKII, Rho-kinase II; EGF, epidermal growth factor; 5-HT, 5-hydroxytryptamine (serotonin); DOCA-salt, deoxycorticosterone acetate salt; LY294002, 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; PSS, physiologic salt solution; PE, phenylephrine; Y27632, (+)-(R)-trans-4-(1-aminoethyl)-N-(4-pyridyl).
- Received October 30, 2003.
- Accepted February 24, 2004.
- The American Society for Pharmacology and Experimental Therapeutics
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