Relaxing and
`Contracting Factors
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`The Endothelium
`Relaxing and Contracting Factors
`Edited by Paul M. Vanhoutte, 1988
`
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`Relaxing
`and
`Contracting
`Factors
`
`Biological and Clinical Research
`
`Edited by
`
`Paul M. Vanhoutte
`
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`Library of Congress Cataloging in Publication Data
`
`Main entry under title:
`
`Relaxing and Contracting Factors.
`
`(Contemporary biomedicine) (The Endothelium)
`Includes bibliographies and index.
`1. Endothelium. 2. Blood-Circulation-Deregulation.
`3. Vasodilators. 4. Blood vessels-Dilatation. 4. Blood
`vessels-Contraction. I. Vanhoutte, Paul M. II. Series.
`m. Series: Endothelium.
`[DNLM: 1. Endothelium-physiol(cid:173)
`ogy. 2. V asoconstrictor Agents-secretion. 3. Vasodi-
`lator Agents-secretion. QV 150 R382]
`QP88.45.R45 1988
`612'.18
`87-22541
`ISBN-13: 978-1-4612-8939-5
`e-ISBN-13: 978-1-4612-4588-9
`DOI: 10.1007 /978-1-4612-4588-9
`
`© 1988 The Humana Press Inc.
`Softcover reprint of the hardcover 1st edition 1988
`Crescent Manor
`PO Box 2148
`Clifton, NJ 07015
`
`All rights reserved
`
`No part of this book may be reproduced, stored in a retrieval
`system, or transmitted in any form or by any means, electronic,
`mechanical, photocopying, microfilming, recording, or otherwise
`without written permission from the Publisher.
`
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`Preface
`
`It is an exciting task to be the editor of the first monograph covering
`a new area of the biomedical sciences. Since the first report in 1980 by
`Robert Furchgott and colleagues (see Chapter 1) of the evidence of
`endothelium-dependent relaxation in isolated arteries, there are ever(cid:173)
`increasing numbers of vascular physiologists and pharmacologists who are
`scraping away the endothelium to look into its role in cardiovascular con(cid:173)
`trol. And the more one looks, the more one discovers. Not only is the
`list of substances that can induce endothelium-dependent relaxations im(cid:173)
`pressively long, but these intriguing cells can also secrete vasoconstrictor
`substances. The ability of the endothelium to modulate the degree of con(cid:173)
`traction of the underlying smooth muscle is an ancestral property of the
`blood vessel wall, illustrating the logic of nature, since the endothelial
`cells are located in the best possible strategic location to continuously
`monitor the properties ( chemical or physical) of the blood. And more and
`more data emerge suggesting that in several cardiovascular diseases per(cid:173)
`turbations in endothelium-dependent responses are one of the early signs
`of the abnormal process. Thus, the importance of endothelium-dependent
`responses, triggered by the intellectual curiosity of one of the pioneers
`of vascular physiology and pharmacology, is now recognized not only by
`basic scientists, but also by all concerned with the cardiovascular diseases.
`The.purpose of this monograph is to provide them with a reference work,
`so that they know where to start.
`As editor, I recognized the overlapping of information among the
`authors. Because it is crucial, however, at such an early stage of this par(cid:173)
`ticular quest for knowledge to confirm, reconfirm, and extend the original
`observations, and with each author having a different approach, the out(cid:173)
`come is a richer knowledge of this growing field.
`I would like to thank the authors for their exciting chapters. My
`secretaries, Mrs. J. Beckman and Mrs. C. Camrud, deserve my gratitude
`for helping me to coach the effort. And my special thanks goes to Mr.
`Lanigan and his staff, at Humana Press, who masterly converted the edited
`chapters into a publication of the highest technical quality.
`Paul M. Vanhoutte
`
`V
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`Contents
`
`Preface ................................. v
`List of Contributors . . . . . . . . . . . . . . . . . . . . . xxiii
`
`CHAPTER 1
`Endothelium-Dependent Relaxation in Systemic Arteries
`Robert F. Furchgott
`1. Historical Perspective . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
`1.1. The Discovery of Acetylcholine-Induced Endothelium(cid:173)
`Dependent Relaxation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
`1.2. Other Early Findings, Conclusions, and Speculation . . . . 5
`2. Recent Developments ................................. · 10
`2.1. The Role of Cyclic GMP in Endothelium-Dependent
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
`Relaxation
`2.2. Additional Agents Producing Endothelium-Dependent
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
`Relaxation
`2. 3. Inhibitors of Endothelium-Dependent Relaxation . . . . . . . 17
`2.4. Further Characterization of EDRF . . . . . . . . . . . . . . . . . . 18
`References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
`
`CHAPTER 2
`Endothelium-Dependent Contractions in Veins and Arteries
`Paul M. Vanhoutte
`1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
`• • • • • . • • • • • . . • 27
`2. Metabolite(s) of Arachidonic Acid (EDCF1)
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
`2.1. Veins
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
`2.2. Cerebral Arteries
`3. Hypoxia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
`4. Conclusion
`38
`References
`
`vii
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`viii
`
`Contents
`
`CHAPTER 3
`The Nature of Endothelium-Derived Relaxing Factor
`T. M. Griffith and A. H. Henderson
`1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
`2. Physical Properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
`3. Chemical Properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
`3 .1. Experimental Approach . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
`3.2. Inhibition of Antioxidants . . . . . . . . . . . . . . . . . . . . . . . . . 46
`3.3. Metabolism of Arachidonate and EDRF . . . . . . . . . . . . . . 48
`4. Mechanisms of Production . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
`4 .1. Stimulated Release . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
`4.2. Basal Release
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
`5. Endogenous Inactivation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
`5.1. Hemoglobin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
`5.2. Haptoglobin-Hemoglobin Complex . . . . . . . . . . . . . . . . . . 53
`6. Mode of Action and Inferences for the Identity of EDRF . . . 54
`7. Modulation of Vasomotor Responses by Basal Release of EDRF 55
`7 .1. Pharmacological Implications . . . . . . . . . . . . . . . . . . . . . . . 55
`7 .2. Physiological Implications . . . . . . . . . . . . . . . . . . . . . . . . . 56
`7. 3. Pathophysiological Implications . . . . . . . . . . . . . . . . . . . . . 57
`8. EDRF In Vivo . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
`9. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
`References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
`
`CHAPTER 4
`Metabolism of Arachidonic Acid and Release of
`Endothelium-Derived Relaxing Factors
`Roger A. Johns and Michael J. Peach
`1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
`2. Endothelial Cell Metabolism of Arachidonic Acid . . . . . . . . . 66
`2.1. Cyclooxygenase Pathways . . . . . . . . . . . . . . . . . . . . . . . . . 66
`2.2. Lipoxygenase Pathways ..... •...................... 68
`2.3. Cytochrome P-450 Pathways . . . . . . . . . . . . . . . . . . . . . . . 68
`3. Evidence Relating Arachidonate Metabolites to EDRF . . . . . . 70
`3 .1. Inhibition of Arachidonic Acid Metabolism . . . . . . . . . . . 70
`72
`3.2. Involvement of Phospholipase and Calcium Dependence
`3.3. Exogenous Arachidonic Acid Studies . . . . . . . . . . . . . . . . 73
`3.4. Melittin and Endogenous Arachidonate Studies . . . . . . . . 76
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`Contents
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`ix
`
`3 .5. Antioxidants and Nonspecific Radical Scavengers . . . . . . 77
`3. 6. Electron Spin Resonance Spectroscopy . . . . . . . . . . . . . . . 79
`4. Evidence Against Arachidonate Metabolite Hypothesis . . . . . 80
`4.1. Nonspecific Action of Many Blockers . . . . . . . . . . . . . . . 80
`4.2. Other Fatty Acids Cause Endothelium-Dependent
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81
`Relaxation
`4.3. Possible Hydrophilic Nature of EDRF . . . . . . . . . . . . . . . 82
`4.4. Exogenous Application of Arachidonate Metabolites . . . 82
`5. Future Studies of Endothelium-Derived Relaxing Factor(s) . . 82
`References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83
`
`CHAPTER 5
`Modulation of the Release and Biological Activity of
`Endothelium-Derived Relaxing Factor by Oxygen(cid:173)
`Derived Free Radicals
`Gabor M. Rubanyl and Paul M. Vanhoutte
`1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91
`2. Direct Actions on Vascular Smooth Muscle . . . . . . . . . . . . . . 93
`3. Adrenergic Neurotransmission . . . . . . . . . . . . . . . . . . . . . . . . . 94
`4. Endothelium-Dependent Relaxations . . . . . . . . . . . . . . . . . . . . . 94
`4.1. Endothelium-Derived Relaxing Factor(s) Is Not Likely
`To Be an Oxygen-Derived Free Radical . . . . . . . . . . . . . 94
`4.2. Hydrogen Peroxide Triggers the Release of Endothelium(cid:173)
`Derived Relaxing Factor(s) . . . . . . . . . . . . . . . . . . . . . . . . 96
`4.3. Hydroxyl Radical Facilitates and Superoxide Anion Inhibits
`Endothelium-Dependent Relaxations to Acetylcholine . . . 97
`4.4. Superoxide Anions Inactivate Endothelium-Derived
`Relaxing Factors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98
`5. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101
`References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102
`
`CHAPTER 6
`Endothelial Cells in Culture and Production of Endothelium(cid:173)
`Derived Relaxing Factor
`T. M. Cocks, J. A. Angus, J. H. Campbell, and G. R. Campbell
`1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 107
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`X
`
`Contents
`
`2. Isolation of EDRF . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 107
`2 .1. Endothelial Cells in Culture . . . . . . . . . . . . . . . . . . . . . . . 108
`2.2. Culture of Endothelial Cells on Microcarrier Beads . . . 114
`2.3. Release of EDRF from Endothelial Cells Grown on
`Microcarrier Beads . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114
`3. Properties of EDRF ................................. 119
`3.1. Half-Life In Vitro: Its Estimation by the Analysis of
`Concentration-Relaxation Curves . . . . . . . . . . . . . . . . . . 119
`3.2. Other Properties of EDRF Released from Cultured
`Endothelial Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126
`4. Whole Artery as a Source of EDRF-A Comparison with
`Cells in Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
`5. Concluding Remarks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131
`References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133
`
`CHAPTER 7
`Endothelial Cells in Culture and Production of Endothelium(cid:173)
`Derived Constricting Factors
`Robert F. Highsmith, David Alchholz, Oliver FltzGerald,
`Richard Paul, Gabor M. Rubanyl, and Kristine Hickey
`
`1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137
`2. Evidence for the Presence of Constricting Factors in
`Vascular Endothelium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 138
`3. Endothelium-Derived Constricting Factor(s) Produced by
`Endothelial Cells in Culture . . . . . . . . . . . . . . . . . . . . . . . . . . 139
`3.1. Methodology for Studying EDCF . . . . . . . . . . . . . . . . . . 139
`3.2. Endothelial Cell Source, Culture Conditions, and Time
`Course of Production of EDCF . . . . . . . . . . . . . . . . . . . . 142
`3.3. Physiological Response to EDCF . . . . . . . . . . . . . . . . . . 143
`3.4. Biochemical and Pharmacological Characterization of
`EDCF......................................... 146
`3.5. Potential Mechanism(s) of Action of EDCF on Vascular
`Smooth Muscle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 149
`4. Effect of Hypoxia on EDCF Release . . . . . . . . . . . . . . . . . . . 150
`5. Concluding Remarks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 156
`References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 157
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`
`CHAPTER 8
`Basal Release of Endothelium-Derived Relaxing Factor
`WIiiiam Martin
`1. Scope . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159
`2. Detection of Basal EDRF Release . . . . . . . . . . . . . . . . . . . . . 159
`2.1. Cascade Bioassay Experiments . . . . . . . . . . . . . . . . . . . . 159
`2.2. Basal EDRF Release and Cyclic GMP . . . . . . . . . . . . . . 160
`2.3. Calcium Dependency . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161
`2.4. Potentiating Action of Phosphodiesterase Inhibitors . . . 161
`3. Endothelium-Dependent Depression of Vasoconstrictor
`Responses
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 164
`3 .1. EDRF Release Elicited by Vasoconstrictor Agents . . . . 165
`3.2. Depression of Alpha-Adrenergic Contractions by
`Spontaneously Released EDRF in the Rat Aorta . . . . . . 166
`3.3. Hemoglobin and Methylene Blue . . . . . . . . . . . . . . . . . . 169
`3.4. Efficacy of Alpha-Adrenergic Agonists . . . . . . . . . . . . . 170
`3.5. Aorta of the Rabbit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 171
`4. Depression of Resting and Stimulated 45C2+ Influx by
`Spontaneously Released EDRF . . . . . . . . . . . . . . . . . . . . . . . . 171
`5. Hypoxia-Induced Endothelium-Dependent Reponses . . . . . . . 171
`6. Differences in Basal EDRF Release . . . . . . . . . . . . . . . . . . . . 172
`7. Basal EDRF Release and Vasospasm . . . . . . . . . . . . . . . . . . . 173
`8. Concluding Remarks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 173
`References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 174
`
`CHAPTER 9
`Calcium Transport Mechanisms in Endothelial Cells
`Regulating the Synthesis and Release of Endothelium(cid:173)
`Derived Relaxing Factor
`Gabor M. Rubanyl, Arnold Schwartz, and Paul M. Vanhoutte
`1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 179
`2. Influx of Extracellular Ca1+ . . . . . . . . . . . . . . . . . . . . . . . . . . . 180
`2 .1. Removal of Extracellular Ca1+ . . . . . . . . . . . . . . . . . . . . . 180
`2.2. Ca1+ Channel Agonists . . . . . . . . . . . . . . . . . . . . . . . . . . . 180
`2. 3. Ca2+ Channel Antagonists . . . . . . . . . . . . . . . . . . . . . . . . 182
`2.4. Na+-ca2+ Exchange . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183
`3. Liberation of Ca1+ from Intracellular Pools . . . . . . . . . . . . . . 184
`4. Concluding Remarks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 184
`References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 186
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`xii
`
`Contents
`
`CHAPTER 10
`Release of Endothelium-Derived Relaxing Factor(s) by
`Physicochemical Stimuli
`Eberhard Bassenge, Rudi Busse, and Ulrlch Pohl
`1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189
`2. EDRF Release and Vascular Response: Asymmetric Behavior
`of the Vascular Wall . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190
`2.1. Bipolar Release . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190
`2.2. Differential Response of Inner and Outer Smooth.Muscle
`to EDRF . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 193
`2. 3. Transmural Endothelial Stimulation . . . . . . . . . . . . . . . . 194
`3. Flow-Rate and Regulation of Vascular Tone . . . . . . . . . . . . . 196
`3 .1. Vasomotion of Arteries in Response to Changes in
`Flow Rate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 196
`3.2. Endothelial Cells as Flow Sensors . . . . . . . . . . . . . . . . . 196
`3.3. Physiological Relevance of Flow-Dependent Dilation . . 199
`3.4. Effect of Long-Term Alterations in Flow Rate on
`Vascular Caliber . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 200
`4. Pulsatile Flow and Release of Endothelium-Derived
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 201
`Vasodilators
`4.1. Prostacyclin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 201
`4.2. Endothelium-Derived Relaxant Factor . . . . . . . . . . . . . . 202
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 203
`5. Oxygen
`5 .1. Hypoxia Vs Anoxia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 203
`5.2. Oxygen Sensitivity of the Vascular Wall . . . . . . . . . . . . 204
`5.3. Oxygen and Release/Transfer of Vasoactive Compounds
`from Endothelial Cells .......................... :. 205
`6. Concluding Remarks
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 210
`References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 211
`
`CHAPTER 11
`Role of Cyclic GMP in Endothelium-Dependent Relaxation
`of Vascular Smooth Muscle
`Robert M. Rapoport and Farid Murad
`1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 219
`2. Elevation of Cyclic GMP Levels in Smooth Muscle by Agents
`that Act on the Endothelium . . . . . . . . . . . . . . . . . . . . . . . . . . 220
`3. Effects of Inhibitors of Guanylate Cyclase on Endothelium(cid:173)
`Dependent Relaxation and Formation of Cyclic GMP . . . . . 222
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`Contents
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`xiii
`
`3 .1. Effects of Free Radical Scavengers and Reducing Agents 222
`3.2. Effects of Cyanide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 225
`4. Effects of Inhibitors of Phosphodiesterase on Endothelium(cid:173)
`Dependent Relaxation and Formation of Cyclic GMP . . . . . 225
`5. Effects of Inhibitors of Phospholipase A2 , Cyclooxygenase, and
`Lipoxygenase on Endothelium-Dependent Relaxation and
`Levels of Cyclic GMP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 226
`6. Effects of Inhibitors of Na+ ,K+ Pump and Membrane Depolar(cid:173)
`izing Agents on Endothelium-Dependent Relaxation and
`Formation of Cyclic GMP . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227
`7. Effects of Nitroglycerin-Induced Desensitization on
`Endothelium-Dependent Relaxation and Formation of
`Cyclic GMP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 228
`8. Effects of Contractile Agents on Cyclic GMP Levels and the
`Role of the Endothelium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 229
`9. Role of the Endothelium and Cyclic GMP in the Regulation
`of Basal Tone . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 231
`10. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 232
`References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 233
`
`CHAPTER 12
`Modulation by the Endothelium of Agonist-lnduced
`Contractions of Vascular Smooth Muscle
`R. C. MIiier, V. Schlnl, and P. Schoeffler
`1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 241
`2. Modulatory Effect of Endothelium on Agonist-lnduced
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 241
`Contractile Responses
`3. Dependence of Endothelium-Modulated Responses on Calcium 249
`4. Calcium Channels and Endothelial Cells . . . . . . . . . . . . . . . . 250
`5. Possible Importance of Na+-ca2+ Exchange for Release
`of EDRF .......................................... 252
`6. Effect of Endothelium on Mobilization of Intracellular
`Calcium for Contraction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 252
`7. Effect of Endothelium on Calcium Uptake by Vascular Tissues 253
`7.1. Basal Uptake . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 253
`7 .2 Stimulated Uptake . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 254
`8. Effect of Endothelium on Efflux of Calcium . . . . . . . . . . . . . 256
`8 .1. Basal Efflux . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 256
`8.2. Efflux of Calcium from Stimulated Tissues . . . . . . . . . . 256
`
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`Contents
`
`9. Effects of Muscarinic Agonists on Ca2• Influx and Efflux in
`Vessels With and Without Endothelium . . . . . . . . . . . . . . . . . 257
`10. Importance of the Modulatory Effect of Endothelium on
`Cellular Ca2
`• Movements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 258
`11. Do Contractile Agonists Stimulate the Release of EDRF or
`Are Modulatory Effects Dependent on Basal Release of EDRF? 258
`References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 260
`
`CHAPTER 13
`Endothelium-Derived Relaxing Factor Relaxes Vascular
`Smooth Muscle by Cyclic GMP-Mediated Effects on
`Calcium Movements
`P. Collins, M. J. Lewis, and A. H. Henderson
`1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 267
`2. Calcium Influx . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 268
`2.1. EDRF . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 268
`2.2. Nitrovasodilators, 8-Bromo-cyclic GMP . . . . . . . . . . . . . 269
`2.3. Cyclic GMP and Calcium Influx . . . . . . . . . . . . . . . . . . . 269
`2.4. Specificity for ''Receptor-Operated Channels''? . . . . . . 271
`3. Calcium Efflux . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 273
`3.1. EDRF . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 273
`3.2. Nitrovasodilators, 8-Bromo-cyclic GMP . . . . . . . . . . . . . 275
`3.3. Implications of Efflux Data . . . . . . . . . . . . . . . . . . . . . . . 275
`4. Biochemical Mechanisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 277
`5. Concluding Remarks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 278
`References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 279
`
`CHAPTER 14
`Heterogeneity in Endothelium-Dependent Relaxations:
`Acute, Chronic, and Evolutionary Modulations
`Virginia M. MIiier and Paul M. Vanhoutte
`1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 285
`2. Hormones
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 286
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`2 .1. Acute Effects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 286
`2.2. Chronic Effects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 289
`3. Innervation
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 292
`4. Blood Flow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 292
`4 .1. Acute Conditions
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 295
`4.2. Chronic Conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 296
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 298
`5. Oxygen
`5 .1. Acute Effects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 298
`5.2. Chronic Effects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 299
`6. Concluding Remarks
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 300
`References . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . 301
`
`CHAPTER 15
`Endothelium-Dependent Regulation of Resting Levels of
`Cyclic GMP and Cyclic AMP and Tension in Pulmonary
`Arteries and Veins
`Louis J. lgnarro, Russell E. Byrns, and Keith S. Wood
`1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 309
`2. Resting Levels of Cyclic GMP and Cyclic AMP . . . . . . . . . 311
`3. Endothelium-Dependent Regulation of Vascular Cyclic GMP
`Levels
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 314
`3.1. Methylene Blue . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 314
`3.2. MB 22948 . . . . . . . . . . . . . . . . . . . . . . . . . . . . ... . . . . . . . 314
`4. Dependence of Activity of EDRF on the Diameter of the
`Blood Vessels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 316
`4.1. Artery
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 316
`4.2. Vein . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 319
`5. Relationship Between Resting Cyclic Nucleotide Levels and
`Endothelial Integrity in Arteries and Veins . . . . . . . . . . . . . . 320
`6. Some Unique Properties of Putative EDRF . . . . . . . . . . . . . . 322
`6.1. Direct Activation of Soluble Guanylate Cyclase . . . . . . 322
`6.2. Stability of EDRF . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 322
`6.3. Chemical Reactivity of EDRF . . . . . . . . . . . . . . . . . . . . . 325
`6.4. Sites of Action of Agents Affecting EDRF Activity . . . 325
`7. Conclusions and Future Direction . . . . . . . . . . . . . . . . . . . . . . 326
`References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 329
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`CHAPTER 16
`Endothelium-Dependent Responses of Cerebral Arteries
`Z. S. Katuslc and J. T. Shepherd
`1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 333
`2. Endothelium-Dependent Relaxations . . . . . . . . . . . . . . . . . . . . 333
`2.1. Acetylcholine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 333
`2.2. Bradykinin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 334
`2.3. Thrombin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 334
`2.4. Vasopressin and Oxytocin . . . . . . . . . . . . . . . . . . . . . . . . 334
`3. Endothelium-Dependent Contractions . . . . . . . . . . . . . . . . . . . 339
`3 .1. Arachidonic Acid, Calcium lonophore A23187, and
`Acetylcholine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 339
`3.2. Anoxia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 339
`3. 3. Potassium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 340
`3.4. Stretch
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 341
`References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 343
`
`CHAPTER 17
`Endothelium, Blood Flow, and Vascular Responses in
`Large Coronary and Iliac Arteries of the Conscious Dog
`Mark A. Young and Stephen F. Vatner
`1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34 7
`2. Direct, Endothelium-Independent Vasodilatation . . . . . . . . . . 348
`3. Endothelium-Mediated, Blood Flow-Independent Vasodilatation 349
`4. Endothelium-Dependent, Blood Flow-Mediated Vasodilatation 352
`5. Endothelium-Mediated Protection Against Vasoconstriction . 355
`6. Concluding Remarks
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 356
`References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 358
`
`CHAPTER 18
`Endothelium-Dependent Responses in Large Arteries and in
`the Microcirculation
`J. A. Angus, T. M. Cocks, C. E. Wright, K. Satoh, and
`G. R. Campbell
`1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 361
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`
`2. EDRF and Reactivity of Large Arteries In Vivo . . . . . . . . . 362
`2.1. Sonomicrometry in Femoral and Coronary Arteries
`under Conditions of Controlled Flow and Pressure . . . . 362
`2.2. Chronic Measurement of the Diameter of the Carotid
`Artery after Removal of the Endothelium . . . . . . . . . . . 364
`3. Reactivity of Large Arteries In Vitro . . . . . . . . . . . . . . . . . . . 368
`3.1. EDRF Is Released by Norepinephrine and- Serotonin . . 368
`3.2. Classification of Endothelial Alpha2-Adrenoceptors . . . 370
`3.3. Distribution of Alpha2-Adrenoceptors on Endothelium . 370
`3.4. Comparison of Endothelium-Dependent Agonists in
`Five Large Arteries . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 373
`3.5. Carotid Artery Reactivity 4 wk after Endothelial
`Denudation
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 374
`4. Reactivity of Microcirculation to EDRF . . . . . . . . . . . . . . . . 379
`4.1. Effect of Hypertension on Reactivity of the Hindquarter
`of the Rabbit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 379
`4.2. Effect of Cholesterol on EDRF . . . . . . . . . . . . . . . . . . . . 379
`5. EDRF and Coronary Atheroma . . . . . . . . . . . . . . . . . . . . . . . 380
`6. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 385
`References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 386
`
`CHAPTER 19
`Endothelium-Dependent Responses in the Peripheral
`Microcirculation
`T. M. Griffith, D. H. Edwards, R. L. Davies, T. J. Harrison,
`and K. T. Evans
`1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 389
`2. Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 392
`2 .1. Preparation
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 392
`2.2. X-Ray Techniques. Contrast Medium . . . . . . . . . . . . . . . 392
`2.3. Quantitation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 393
`2.4. Agents Used To Induce Tone. Hemoglobin . . . . . . . . . . 395
`3. Network Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 395
`4. Results and Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 396
`4.1. Basal and Stimulated Release of EDRF. Evidence for a
`Complex Interaction with Vessel Tone . . . . . . . . . . . . . . 396
`4.2. Regulation of Resistance by Basal EDRF Release . . . . . 402
`4.3. Flow-Dependent EDRF-Mediated Dilatation . . . . . . . . . 402
`4.4. Regulation of Flow Distribution by Basal EDRF Release 405
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`4.5. Influence of Basal EDRF Release on Power Losses and
`Pressure-Flow Characteristics . . . . . . . . . . . . . . . . . . . . . 407
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 O
`5. Concluding Remarks
`References . . . . . . . . . . . . . . . . . . . . . . . . . .