Published online before print December 16, 2002, doi: 10.1161/01.HYP.0000047240.33861.6B
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(Hypertension. 2003;41:801.)
© 2003 American Heart Association, Inc.
Scientific Contributions |
20-HETE–Induced Contraction of Small Coronary Arteries Depends on the Activation of Rho-Kinase
From Institut für Kardiovaskuläre Physiologie, Klinikum der J.W.G.-Universität, Frankfurt am Main, Germany.
Correspondence to Ingrid Fleming, PhD, Institut für Kardiovaskuläre Physiologie, Klinikum der J.W.G.-Universität, Theodor-Stern-Kai 7, D-60590 Frankfurt am Main, Germany. E-mail fleming{at}em.uni-frankfurt.de
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Abstract |
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20-HETE is a potent constrictor of small blood vessels and has been suggested to play a crucial role in the generation of myogenic tone and the development of hypertension. In the present study, we investigated the mechanisms by which exogenously applied 20-HETE modulates vascular tone in small porcine coronary arteries. In organ chamber experiments, 20-HETE elicited a concentration-dependent contraction of small porcine coronary artery rings that was partially inhibited by the cyclooxygenase inhibitor diclofenac, the thromboxane and endoperoxide receptor antagonist SQ29548, and the thromboxane A2 synthase inhibitor furegrelate. Removal of endothelium attenuated the response to 20-HETE, whereas preconstriction of endothelium-denuded vessels to 25% of the maximum response with KCl markedly enhanced the response to 20-HETE. This 20-HETE–induced contraction was not associated with a significant increase in the intracellular concentration of Ca2+. 20-HETE–induced contraction was also observed in ß-escin–permeabilized arteries precontracted with a submaximal concentration of Ca2+ and was abolished by the Rho-kinase inhibitor Y27632, but was insensitive to the PKC inhibitor RO 31-8220. 20-HETE elicited the phosphorylation of the myosin light chain (MLC20) in coronary artery rings, an effect that was sensitive to Y27632 and mimicked by the thromboxane analog U46619. These data suggest that in small porcine coronary arteries, 20-HETE can induce contraction by 2 mechanisms, one endothelium-dependent involving the cyclooxygenase-dependent generation of vasoconstrictor prostanoids, and the other endothelium-independent. The latter response is associated with the activation of Rho-kinase, phosphorylation of MLC20, and sensitization of the contractile apparatus to Ca2+.
Key Words: 20-HETE • kinase • myosin • calcium • arteries
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Introduction |
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As a potent vasoconstrictor, 20-HETE, an
-hydroxylation product of arachidonic acid catalyzed by cytochrome P450 (CYP) 4A enzymes, is an essential component of the signal transduction cascade activated by several hormonal systems that have central roles in blood pressure regulation (for reviews, see Capdevila and Falck1 and Roman2).
Initially, 20-HETE was reported to affect vascular smooth muscle tone indirectly as a result of its metabolism by cyclooxygenase in endothelial cells, inasmuch as indomethacin and the endoperoxide/thromboxane receptor antagonist SQ29548 inhibited endothelium-dependent contraction.3–5 The inhibitory effect of SQ29548, the appearance of labile metabolites with a half-life of 
6 minutes, and the production of 20-hydroxy-PGF2
by SnCl2 reduction indicated that the vasoconstrictor metabolites of 20-HETE are the labile endoperoxides of 20-HETE, 20-hydroxy-PGG2, and 20-hydroxy-PGH2.3,5
Although the effects of 20-HETE in large arteries and in the aorta are at least partially cyclooxygenase-dependent, data gathered over the past 5 years have convincingly demonstrated that 20-HETE is generated endogenously within vascular smooth muscle cells from small resistance arteries. 20-HETE is synthesized in response to an increase in [Ca2+]i elicited by stimuli such as an increase in transmural pressure6–11 and the vasoconstrictor agonists endothelin-112–16 and angiotensin II.17–21 Once formed, 20-HETE is thought to increase smooth muscle tone by inhibiting large conductance Ca2+-activated K+ (BK) channels,22,23 inducing depolarization and further increasing [Ca2+]i,7 an effect that has been attributed to the activation of L-type Ca2+ channels10,24 and/or the activation of PKC and inhibition of the Na-K-ATPase.25–27
Because 20-HETE is thought to mediate myogenic tone in resistance-sized arteries, and a Rho-kinase–dependent mechanism contributes to cerebral vascular tone in vivo,28 we speculated that 20-HETE may elicit contraction, not only by increasing [Ca2+]i but also by increasing the sensitivity of contractile proteins to Ca2+ via a Rho-kinase–dependent mechanism. The aim of the present study was therefore to determine the mechanisms by which exogenously applied 20-HETE modulates vascular tone in small porcine coronary arteries.
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Methods |
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Vascular Reactivity Studies
Second branches of coronary arteries (internal diameter, 300 to 500 µm) were dissected from the hearts of freshly slaughtered pigs, cleaned of adventitial adipose and connective tissue, and cut into 4-mm-long segments. In some experiments, the endothelium was removed by intraluminal perfusion with CHAPS (0.5% in Tyrode’s solution) for 30 seconds.
Coronary artery rings were mounted on stainless-steel triangles connected to a force transducer (Hugo Sachs Elektronik-Harvard Apparatus) and a rigid support for measurement of isometric force in organ baths containing Tyrode’s solution of the following composition (in mmol/L): NaCl 132, KCl 4, CaCl2 1.6, MgCl2 0.98, NaHCO3 23.8, NaH2PO4 0.36, glucose 10, and Ca-Titriplex 0.05. The solution was gassed with 20% O2, 5% CO2, and 75% N2 to give a pO2 of 140 mm Hg and pH 7.4 at 37°C. Passive tension was gradually adjusted over a 60-minute period to 1 g, thereafter arterial rings were repeatedly exposed to a modified Tyrode’s solution rich in KCl (80 mmol/L) until stable contractions were obtained. The presence of functional endothelium was assessed in all preparations by the ability of bradykinin (1 µmol/L) to induce relaxation of vessels precontracted with U46619 (0.1 µmol/L), and vessels exhibiting <80% relaxation were discarded.
Assay of Thromboxane A2 Production by Porcine Coronary Endothelial Cells
Porcine coronary endothelial cells were isolated as described29 and seeded onto 48-well culture plates. Six hours after seeding, the cell supernatant was replaced with Tyrode’s solution (150 mL), and the cells were stimulated with solvent (0.1% ethanol), 20-HETE (1 µmol/L), or ionomycin (0.1 µmol/L). After a further 10 minutes, the supernatant was removed, and thromboxane B2 (TXB2, a stable metabolite of thromboxane A2 [TXA2]) levels were assayed with a commercially available kit (PerkinElmer Life Sciences).
Measurement of [Ca2+]i
Contraction and [Ca2+]i were measured simultaneously. Contraction was measured as described above, and changes in [Ca2+]i were determined by measuring changes in fura-2 fluorescence. For the measurement of [Ca2+]i, arterial rings were mounted between 2 tungsten wires (30 µm diameter) connected to a rigid support attached to a micromanipulator and a force transducer, superfused with modified Tyrode’s solution, and placed on the stage of an inverted microscope (Diaphot-TMB; Nikon). Vessels were loaded with fura-2 by incubating with modified Tyrode’s solution containing fura-2/AM (5 µmol/L) and pluronic F-127 (0.025% wt/vol) at room temperature for 120 minutes. [Ca2+]i was determined fluorometrically by continuous rapid alternating excitation from dual monochromators set at 340 and 380 nm (Deltascan, Photon Technology) as described.30 At the end of each experiment, the vessels were superfused with a buffer containing CaCl2 (5 mmol/L), ionomycin (1 µmol/L), and KCl (80 mmol/L). After a stable 340:380 ratio was achieved, the buffer was changed to a Ca2+-free one. The background fluorescence was calculated after the addition of MnCl2 (10 mmol/L) and subtracted from the original signals. Data are presented as changes in the 340/380 ratio. Using this method, the fluorescence (340/380 nm) was 1.4±0.05 under basal conditions and 3.4±1.3 in response to ionomycin, CaCl2 and KCl.
Permeabilization of the Small Porcine Coronary Arteries
Arteries were permeabilized using ß-escin as described,31 with minor modifications. Briefly, arteries were incubated at room temperature for 45 minutes in PIPES solution (20 mmol/L PIPES, 10 mmol/L creatine phosphate, 5.2 mmol/L Na2ATP, 5.1 mmol/L MgCl2, 87 mmol/L KCl, 1 µmol/L leupeptin, 1 µmol/L ionomycin, 1 µmol/L calmodulin, and 4 mmol/LEGTA at pH 7.4) supplemented with ß-escin (50 µmol/L). Arteries were then mounted in the organ chamber containing PIPES buffer (37°C). Passive tension was adjusted to 1 g as described above. After a short equilibration period, the buffer was replaced with one of a similar composition containing 2 mmol/L EGTA and contraction elicited with CaCl2 (10 µmol/L). After washing, the arteries were re-equilibrated in the PIPES buffer, and a submaximal concentration of Ca2+ (1 µmol/L) was added. When the contraction reached a plateau, 20-HETE (1 µmol/L) and GTP (10 µmol/L) were also applied.
Immunoblotting
Endothelium-intact rings of porcine coronary artery were incubated for 30 minutes at 37°C in modified Tyrode’s solution and stimulated as described in Results. Vessels were then frozen in liquid nitrogen, ground to a powder, and suspended in trichloroacetic acid (10% w/v). After 30 minutes at 4°C, the suspension was centrifuged (4°C) at 14 000g for 30 minutes, the supernatant was discarded, and the pellet washed 3 times with water-saturated diethyl ether. Air-dried samples were solubilized in Triton X-100 lysis buffer, and proteins in the supernatant (50 µg) were heated with sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) sample buffer, separated by SDS-PAGE (12%), and transferred to a nitrocellulose membrane as described.32 Membranes were cut at the 32-kDa marker, and ß-actin and myosin light chain 20 (MLC20) were detected by selective monoclonal antibodies (Sigma) and visualized by enhanced chemiluminescence using a commercially available kit (Amersham).
Statistical Analysis
Contractions are expressed in grams of tension or as a percentage of the maximal contraction (% max) obtained using 80 mmol/L KCl. Data are expressed as mean±SEM, and statistical evaluation was performed with the Student t test for unpaired data or 1-way ANOVA followed by a Bonferroni t test, when appropriate. Values of P<0.05 were considered statistically significant.
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Effect of 20-HETE on the Tone of Endothelium-Intact Rings of Small Porcine Coronary Artery
Application of 20-HETE (0.1 to 1 µmol/L) to endothelium-intact rings of small porcine coronary arteries resulted in a concentration-dependent contraction (Figure 1). The kinetics of the response depended on the concentration applied, as the increase in tone observed after the application of lower concentrations of 20-HETE developed slowly, whereas the contraction observed after application of the highest concentration was rapid.
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The 20-HETE–induced contraction was significantly attenuated by the cyclooxygenase inhibitor diclofenac (10 mmol/L; Figures 1B and 1C), as well as the TXA2 synthase inhibitor furegrelate (10 µmol/L) and the TXA2/PGH2 receptor antagonist SQ-29548 (10 µmol/L; Figure 1C).
To assess whether 20-HETE elicited the generation of TXA2 by porcine coronary endothelial cells, we determined the release of TXB2 from freshly isolated cells stimulated with 20-HETE. 20-HETE (1 µmol/L) elicited a small increase in TXB2 levels; levels were 2.92±0.09, 4.26±0.27, and 8.37±0.72 pg TXB2/mg protein after stimulation with solvent, 20-HETE, and ionomycin, respectively (P<0.01, n=6).
Effect of 20-HETE on the Tone of Endothelium-Denuded Rings of Small Porcine Coronary Artery
Endothelial denudation markedly attenuated the contractile response to 20-HETE, so that an increase in tone was only observed with the highest concentration used (Figure 2). However, a larger concentration-dependent increase in tone was observed when 20-HETE was applied to rings that were precontracted with KCl to 25% of the maximal contraction to 80 mmol/L KCl (Figure 2). The 20-HETE–induced increase in tone was slightly attenuated by the PKC inhibitor RO 31-8220 (300 nmol/L), but abolished by the Rho-kinase inhibitor Y27632 (1 µmol/L; Figure 2). On its own, Y27632 did not affect the contraction elicited by either 25 or 80 mmol/L KCl (data not shown). In the absence of a functional endothelium, diclofenac and furegrelate did not significantly inhibit the contractile response to 20-HETE (data not shown), whereas SQ-29548 attenuated the 20-HETE (1 µmol/L)–induced contraction by 56.4±0.3% (P<0.01, n=4 to 6).
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Effect of 20-HETE on Vascular Smooth Muscle Cell [Ca2]i
To determine whether or not the 20-HETE–induced increase in tone was associated with an increase in smooth muscle [Ca2+]i, small porcine coronary arteries were loaded with the Ca2+-sensitive fluorescent indicator, fura-2. The application of KCl to coronary artery rings elicited a contraction which was associated with an increase in [Ca2+]i. Although the subsequent addition of 20-HETE further increased contraction, there was no significant corresponding increase in [Ca2+]i (Figure 3).
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Because 20-HETE is reported to elicit contraction by inhibiting BK channels,22,23 depolarizing vascular smooth muscle cells, and opening L-type Ca2+ channels,24 we determined the effect of 20-HETE on the vascular tone of porcine coronary arteries precontracted with KCl in the absence and presence of the BK blocker iberiotoxin and the Ca2+ antagonist, nifedipine. Iberiotoxin (100 nmol/L) did not affect either the basal tone or the 20-HETE-induced contraction of porcine coronary arteries precontracted with KCl (contractions were 45.2±11.9 versus 43.3±8.5% max in the absence and presence of iberiotoxin, respectively; n=4, P=0.77). Significantly higher concentrations of KCl were required to precontract nifedipine-treated coronary artery rings to 25% of the maximal contraction (30 mmol/L versus 80 mmol/L KCl in solvent and nifedipine-treated rings, respectively). The presence of nifedipine, however, did not significantly affect the maximal increase in tone that developed in response to the addition of 20-HETE (contractions were 50.0±6.8 versus 41.5±9.04% max in the absence and presence of nifedipine, respectively; n=6, P=0.46).
Effect of 20-HETE on the Tone of Permeabilized Arteries
Because the ability of 20-HETE to increase the tone of small porcine coronary arteries was markedly enhanced by preconstriction with KCl, but independent of a further increase in [Ca2+]i, we assessed the ability of 20-HETE to increase tone in ß-escin–permeabilized arteries. After permeabilization, arteries were contracted using a submaximal concentration of Ca2+ (1 µmol/L), and once a stable contraction level was reached, 20-HETE and GTP were added to the vessel chamber. The addition of GTP (10 µmol/L) alone was without effect (data not shown), whereas the addition of 20-HETE and GTP elicited a rapid contraction (Figure 4). This contractile response was not significantly affected by RO 31-8220 but was abolished by Y27632 (Figure 4). Identical results were obtained in endothelium-intact and endothelium-denuded arterial rings.
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contraction %) elicited by the application of 20-HETE. Experiments were performed in the absence (CTL) and presence of RO 31-8220 (RO-31; 300 nmol/L) and Y27632 (Y27; 10 µmol/L), each preincubated with the vessel for 30 minutes before contraction with calcium. Results are presented as mean±SEM of data obtained in 5 separate experiments; **P<0.01 vs control (CTL).
Effect of 20-HETE on MLC20 Phosphorylation
Because Y27632 inhibited the contraction to 20-HETE and Ca2+ sensitization in several different arteries has been linked to phosphorylation of MLC20, we determined the effects of 20-HETE on the phosphorylation of MLC20. In extracts of unstimulated porcine coronary artery, MLC20 was detected as a single band (Figure 5). Stimulation of coronary arteries with 20-HETE (1 µmol/L) induced a shift in the mobility of this protein, which was not observed in extracts from arteries treated with 20-HETE and Y27632. A similar phenomenon was observed in coronary arteries stimulated with the known Ca2+-sensitizing agent U46619 (Figure 5).
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Discussion |
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The results of the present investigation indicate that in small porcine coronary arteries, 20-HETE enhances vascular tone by a mechanism involving the activation of Rho-kinase, phosphorylation of MLC20, and the sensitization of the contractile apparatus to Ca2+. When the vascular endothelium was intact, the contractile response appeared to be dependent on the metabolism of 20-HETE by cyclooxygenase, whereas the latter enzyme was not involved in the contractile response to 20-HETE in endothelium-denuded arteries.
The metabolism of 20-HETE in endothelial cells by cyclooxygenase has been appreciated for a number of years, and although we found that 20-HETE elicited the generation of TXB2 from freshly isolated endothelial cells, it is most likely that the endothelium-dependent contractile response obtained can be attributed to the labile endoperoxides, 20-hydroxy-PGG2 and 20-hydroxy-PGH2.3–5
Although 20-HETE is reported to be a potent constrictor of isolated endothelium-denuded renal skeletal muscle and cerebral arteries, contraction of endothelium-denuded small porcine coronary arteries to 20-HETE was only observed when high concentrations were used. The contractile response was, however, markedly potentiated when vessels were precontracted to 25% of the maximal KCl-inducible tone, suggesting that 20-HETE may potentiate contraction by sensitizing the contractile proteins to the increase in [Ca2+]i induced by KCl.
20-HETE is reported to induce vascular smooth muscle contraction by inhibiting BK channels,22,23 thus depolarizing the membrane and increasing Ca2+ entry via L-type Ca2+ channels.10,24 However, in the present investigation, we found no evidence linking 20-HETE to this mechanism, as the contractile response to 20-HETE was not affected by the BK channel blocker iberiotoxin. 20-HETE had also no direct effect on Ca2+ signaling as, although we observed a clear increase in smooth muscle [Ca2+]i after the stimulation of arteries with KCl, the subsequent application of 20-HETE did not further increase [Ca2+]i. Moreover, blocking Ca2+ entry via L-type Ca2+ channels with nifedipine failed to affect the contractile response to 20-HETE. In the small porcine coronary artery, therefore, 20-HETE induces contraction chiefly by increasing vascular smooth muscle sensitivity to Ca2+. The best evidence for a 20-HETE–induced Ca2+ sensitization came from experiments using ß-escin–permeabilized arteries, in which [Ca2+]i was clamped at a constant value. Because, under the experimental conditions used, contraction was unrelated to a change in [Ca2+]i, the 20-HETE–induced contraction can be attributed to an increase in the sensitivity of the contractile apparatus to Ca2+. In general, small porcine coronary arteries respond poorly to Ca2+-elevating agonists such as norepinephrine and phenylephrine but are sensitive to the TXA2 analog U46619. Indeed, the plateau phase of the U46619-induced contraction is achieved mainly by enhancing the sensitivity of the contractile apparatus to a relatively small increase in [Ca2+]i by a mechanism involving the activation of Rho-kinase.33
As PKC-mediated and Rho-kinase–mediated Ca2+ sensitization mechanisms are operative in small coronary arteries,33 we determined the effects of PKC and Rho-kinase inhibitors on the 20-HETE–induced increase in coronary artery tone. The PKC inhibitor, RO 31-8220, did not significantly affect the 20-HETE–induced contraction of endothelium-denuded arteries, whereas the response was abolished by the Rho-kinase inhibitor Y27632. A similar abolition of the 20-HETE–induced contraction in the presence of Y27632 was also observed in ß-escin–permeabilized arteries.
Although the intracellular mechanisms underlying the phenomenon of calcium sensitization have not been fully elucidated, numerous studies have shown that it is accompanied by the phosphorylation of MLC20.34 The extent of MLC20 phosphorylation is determined by the activity of the myosin light chain kinase and the myosin light-chain phosphatase, such that an increase in the activity of the MLC kinase and/or a decrease in the activity of the phosphatase increases MLC20 phosphorylation (for reviews, see Somlyo and Somlyo34 and Fukata35). PKC and Rho-kinase can both sensitize vascular smooth muscle to Ca2+, although the molecular mechanisms eventually leading to enhanced MLC20 phosphorylation are distinct; Rho-kinase phosphorylates and inhibits myosin light chain phosphatase, leading to an increased MLC20 phosphorylation and contraction,36,37 whereas PKC phosphorylates CPI-17, a myosin light chain phosphatase inhibitor.38 In the present study, we observed that 20-HETE induced the phosphorylation of MLC20 (assessed as a mobility shift in Western blotting experiments) in small porcine coronary arteries. This response, like the potentiation of contraction, was abolished in arteries preincubated with the Rho-kinase inhibitor. A similar effect was also observed in arteries treated with U46619, which also sensitizes small porcine coronary arteries to Ca2+ via the activation of Rho-kinase.33 Taken together, the results of the present investigation indicate that 20-HETE is able to sensitize small porcine coronary artery smooth muscle cells to Ca2+ by activating Rho-kinase and enhancing the phosphorylation of MLC20.
Perspectives
20-HETE is currently characterized as a prohypertensive eicosanoid and has the potential to play a dual role in the regulation of blood pressure by virtue of its ability to induce contraction, as well as to inhibit sodium reabsorption. Several reports have demonstrated that the expression of CYP 4A and the production of 20-HETE are altered in genetic and experimental models of hypertension (for review, see Capdevila and Falck1 and Roman2). It is therefore tempting to speculate that some of the potentially beneficial effects of Rho-kinase inhibitors in hypertension28,39 may be related to the inhibition of 20-HETE–induced Ca2+ sensitization. Moreover, although a direct role for 20-HETE in coronary artery vasospasm has not been described to-date, the fact that Rho-kinase inhibitors effectively reduce vasospasm in porcine models,40,41 as well as in patients with vasospastic angina,42 may be indicative of a role for this eicosanoid in vasospasm.
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Acknowledgments |
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Research described in this article was supported by Philip Morris Inc. We are indebted to Tanja-Maria Mareczek for expert technical assistance.
Received October 4, 2002; first decision October 24, 2002; accepted November 4, 2002.
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References |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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1. Capdevila JH, Falck JR. The CYP P450 arachidonic acid monooxygenases: from cell signaling to blood pressure regulation. Biochem Biophys Res Commun. 2001; 285: 571–576.[CrossRef][Medline] [Order article via Infotrieve]
2. Roman RJ. P-450 metabolites of arachidonic acid in the control of cardiovascular function. Physiol Rev. 2002; 82: 131–185.
3. Schwartzman ML, Falck JR, Yadagiri P, Escalante B. Metabolism of 20-hydroxyeicosatetraenoic acid by cyclooxygenase: formation and identification of novel endothelium-dependent vasoconstrictor metabolites. J Biol Chem. 1989; 264: 11658–11662.
4. Escalante B, Sessa WC, Falck JR, Yadagiri P, Schwartzman M. Vasoactivity of 20-hydroxyeicosatetraenoic acid is dependent on metabolism by cyclooxygenase. J Pharmacol Exp Ther. 1989; 248: 229–232.
5. Escalante B, Omata K, Sessa W, Lee SG, Falck JR, Schwartzman ML. 20-Hydroxyeicosatetraenoic acid is an endothelium-dependent vasoconstrictor in rabbit arteries. Eur J Pharmacol. 1993; 235: 1–7.[CrossRef][Medline] [Order article via Infotrieve]
6. Kauser K, Clark JE, Masters BS, Ortiz de Montellano PR, Ma YH, Harder DR, Roman RJ. Inhibitors of cytochrome P-450 attenuate the myogenic response of dog renal arcuate arteries. Circ Res. 1991; 68: 1154–1163.
7. Ma YH, Gebremedhin D, Schwartzman ML, Falck JR, Clark JE, Masters BS, Harder DR, Roman RJ. 20-Hydroxyeicosatetraenoic acid is an endogenous vasoconstrictor of canine renal arcuate arteries. Circ Res. 1993; 72: 126–136.
8. Zou AP, Imig JD, Kaldunski M, Ortiz de Montellano PR, Sui Z, Roman RJ. Inhibition of renal vascular 20-HETE production impairs autoregulation of renal blood flow. Am J Physiol. 1994; 266: F275–F282.[Medline] [Order article via Infotrieve]
9. Imig JD, Zou AP, Stec DE, Harder DR, Falck JR, Roman RJ. Formation and actions of 20-hydroxyeicosatetraenoic acid in rat renal arterioles. Am J Physiol. 1996; 270: R217–R227.[Medline] [Order article via Infotrieve]
10. Harder DR, Lange AR, Gebremedhin D, Birks EK, Roman RJ. Cytochrome P450 metabolites of arachidonic acid as intracellular signaling molecules in vascular tissue. J Vasc Res. 1997; 34: 237–243.[Medline] [Order article via Infotrieve]
11. Gebremedhin D, Lange AR, Lowry TF, Taheri MR, Birks EK, Hudetz AG, Narayanan J, Falck JR, Okamoto H, Roman RJ, Nithipatikom K, Campbell WB, Harder DR. Production of 20-HETE and its role in autoregulation of cerebral blood flow. Circ Res. 2000; 87: 60–65.
12. Oyekan AO, McGiff JC. Cytochrome P-450–derived eicosanoids participate in the renal functional effects of ET-1 in the anesthetized rat. Am J Physiol. 1998; 274: R52–R61.[Medline] [Order article via Infotrieve]
13. Imig JD, Pham BT, LeBlanc EA, Reddy KM, Falck JR, Inscho EW. Cytochrome P450 and cyclooxygenase metabolites contribute to the endothelin-1 afferent arteriolar vasoconstrictor and calcium responses. Hypertension. 2000; 35: 307–312.
14. Hercule HC, Oyekan AO. Cytochrome P450 /-1 hydroxylase-derived eicosanoids contribute to endothelinA and endothelinB receptor–mediated vasoconstriction to endothelin-1 in the rat preglomerular arteriole. J Pharmacol Exp Ther. 2000; 292: 1153–1160.
15. Hercule HC, Oyekan AO. Role of NO and cytochrome P-450–derived eicosanoids in ET-1–induced changes in intrarenal hemodynamics in rats. Am J Physiol. 2000; 279: R2132–R2141.
16. Escalante BA, McGiff JC, Oyekan AO. Role of cytochrome P-450 arachidonate metabolites in endothelin signaling in rat proximal tubule. Am J Physiol. 2002; 282: F144–F150.
17. Chu ZM, Croft KD, Kingsbury DA, Falck JR, Reddy KM, Beilin LJ. Cytochrome P450 metabolites of arachidonic acid may be important mediators in angiotensin II–induced vasoconstriction in the rat mesentery in vivo. Clin Sci (Colch). 2000; 98: 277–282.[Medline] [Order article via Infotrieve]
18. Croft KD, McGiff JC, Sanchez-Mendoza A, Carroll MA. Angiotensin II releases 20-HETE from rat renal microvessels. Am J Physiol. 2000; 279: F544–F551.
19. Sanchez-Mendoza A, Lopez-Sanchez P, Vazquez-Cruz B, Rios A, Martinez-Ayala S, Escalante B. Angiotensin II modulates ion transport in rat proximal tubules through CYP metabolites. Biochem Biophys Res Commun. 2000; 272: 423–430.[CrossRef][Medline] [Order article via Infotrieve]
20. Parmentier JH, Muthalif MM, Nishimoto AT, Malik KU. 20-Hydroxyeicosatetraenoic acid mediates angiotensin II–induced phospholipase D activation in vascular smooth muscle cells. Hypertension. 2001; 37: 623–629.
21. Alonso-Galicia M, Maier KG, Greene AS, Cowley AW, Jr, Roman RJ. Role of 20-hydroxyeicosatetraenoic acid in the renal and vasoconstrictor actions of angiotensin II. Am J Physiol. 2002; 283: R60–R68.
22. Harder DR, Gebremedhin D, Narayanan J, Jefcoat C, Falck JR, Campbell WB, Roman R. Formation and action of a P-450 4A metabolite of arachidonic acid in cat cerebral microvessels. Am J Physiol. 1994; 266: H2098–107.[Medline] [Order article via Infotrieve]
23. Zou AP, Fleming JT, Falck JR, Jacobs ER, Gebremedhin D, Harder DR, Roman RJ. 20-HETE is an endogenous inhibitor of the large-conductance Ca2+-activated K+ channel in renal arterioles. Am J Physiol. 1996; 270: R228–R237.[Medline] [Order article via Infotrieve]
24. Gebremedhin D, Lange AR, Narayanan J, Aebly MR, Jacobs ER, Harder DR. Cat cerebral arterial smooth muscle cells express cytochrome P450 4A2 enzyme and produce the vasoconstrictor 20-HETE which enhances L-type Ca2+ current. J Physiol. 1998; 507: 771–781.
25. Schwartzman M, Ferreri NR, Carroll MA, Songu-Mize E, McGiff JC. Renal cytochrome P450-related arachidonate metabolite inhibits (Na++K+)ATPase. Nature. 1985; 314: 620–622.[CrossRef][Medline] [Order article via Infotrieve]
26. Nowicki S, Chen SL, Aizman O, Cheng XJ, Li D, Nowicki C, Nairn A, Greengard P, Aperia A. 20-Hydroxyeicosa-tetraenoic acid (20 HETE) activates protein kinase C: role in regulation of rat renal Na+,K+-ATPase. J Clin Invest. 1997; 99: 1224–1230.[Medline] [Order article via Infotrieve]
27. Lange A, Gebremedhin D, Narayanan J, Harder DR. 20-Hydroxyeicosatetraenoic acid-induced vasoconstriction and inhibition of potassium current in cerebral vascular smooth muscle is dependent on activation of protein kinase C. J Biol Chem. 1997; 272: 27345–27352.
28. Chrissobolis S, Sobey CG. Evidence that Rho-kinase activity contributes to cerebral vascular tone in vivo and is enhanced during chronic hypertension: comparison with protein kinase C. Circ Res. 2001; 88: 774–779.
29. Popp R, Bauersachs J, Hecker M, Fleming I, Busse R. A transferable, ß-naphthoflavone–inducible, hyperpolarizing factor is synthesized by native and cultured porcine coronary endothelial cells. J Physiol (Lond). 1996; 497: 699–709.
30. Fleming I, Hecker M, Busse R. Intracellular alkalinization induced by bradykinin sustains activation of the constitutive nitric oxide synthase in endothelial cells. Circ Res. 1994; 74: 1220–1226.
31. Kobayashi S, Kitazawa T, Somlyo AV, Somlyo AP. Cytosolic heparin inhibits muscarinic and alpha-adrenergic Ca2+ release in smooth muscle: physiological role of inositol 1,4,5-trisphosphate in pharmacomechanical coupling. J Biol Chem. 1989; 264: 17997–18004.
32. Fleming I, Bauersachs J, Fisslthaler B, Busse R. Ca2+-independent activation of the endothelial nitric oxide synthase in response to tyrosine phosphatase inhibitors and fluid shear stress. Circ Res. 1998; 82: 686–695.
33. Nobe K, Paul RJ. Distinct pathways of Ca2+ sensitization in porcine coronary artery: effects of Rho-related kinase and protein kinase C inhibition on force and intracellular Ca2+. Circ Res. 2001; 88: 1283–1290.
34. Somlyo AP, Somlyo AV. Signal transduction by G-proteins, Rho-kinase and protein phosphatase to smooth muscle and non-muscle myosin II. J Physiol (Lond). 2000; 522: 177–185.
35. Fukata Y, Amano M, Kaibuchi K. Rho-Rho-kinase pathway in smooth muscle contraction and cytoskeletal reorganization of non-muscle cells. Trends Pharmacol Sci.;. 2001; 22: 32–39.[CrossRef][Medline] [Order article via Infotrieve]
36. Toth A, Kiss E, Gergely P, Walsh MP, Hartshorne DJ, Erdodi F. Phosphorylation of MYPT1 by protein kinase C attenuates interaction with PP1 catalytic subunit and the 20 kDa light chain of myosin. FEBS Lett. 2000; 484: 113–117.[CrossRef][Medline] [Order article via Infotrieve]
37. Fu X, Gong MC, Jia T, Somlyo AV, Somlyo AP. The effects of the Rho-kinase inhibitor Y-27632 on arachidonic acid-, GTP
S-, and phorbol ester–induced Ca2+-sensitization of smooth muscle. FEBS Lett. 1998; 440: 183–187.[CrossRef][Medline]
[Order article via Infotrieve]
38. Eto M, Ohmori T, Suzuki M, Furuya K, Morita F. A novel protein phosphatase-1 inhibitory protein potentiated by protein kinase C: isolation from porcine aorta media and characterization. J Biochem (Tokyo). 1995; 118: 1104–1107.
39. Masumoto A, Hirooka Y, Shimokawa H, Hironaga K, Setoguchi S, Takeshita A. Possible involvement of Rho-kinase in the pathogenesis of hypertension in humans. Hypertension. 2001; 38: 1307–1310.
40. Shimokawa H, Seto M, Katsumata N, Amano M, Kozai T, Yamawaki T, Kuwata K, Kandabashi T, Egashira K, Ikegaki I, Asano T, Kaibuchi K, Takeshita A. Rho-kinase–mediated pathway induces enhanced myosin light chain phosphorylations in a swine model of coronary artery spasm. Cardiovasc Res. 1999; 43: 1029–1039.
41. Kandabashi T, Shimokawa H, Miyata K, Kunihiro I, Kawano Y, Fukata Y, Higo T, Egashira K, Takahashi S, Kaibuchi K, Takeshita A. Inhibition of myosin phosphatase by upregulated Rho-kinase plays a key role for coronary artery spasm in a porcine model with interleukin-1ß. Circulation. 2000; 101: 1319–1323.
42. Masumoto A, Mohri M, Shimokawa H, Urakami L, Usui M, Takeshita A. Suppression of coronary artery spasm by the Rho-kinase inhibitor fasudil in patients with vasospastic angina. Circulation. 2002; 105: 1545–1547.
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