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Vascular Function and Structure in the Rat Aorta Vascular Function and Structure in the Rat Aorta Vascular Function and Structure in the Rat Aorta By Keith Wan Kee Ng Vascular Function and Structure in the Rat Aorta, by Keith Wan Kee Ng This book first published 2013 Cambridge Scholars Publishing 12 Back Chapman Street, Newcastle upon Tyne, NE6 2XX, UK British Library Cataloguing in Publication Data A catalogue record for this book is available from the British Library Copyright © 2013 by Keith Wan Kee Ng All rights for this book 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, recording or otherwise, without the prior permission of the copyright owner. ISBN (10): 1-4438-4823-9, ISBN (13): 978-1-4438-4823-7 To My parents My best friends TABLE OF CONTENTS List of Figures............................................................................................. xi List of Tables ............................................................................................ xiii Acknowledgements ................................................................................... xv Preface ..................................................................................................... xvii List of Acronyms ...................................................................................... xix Chapter One ................................................................................................. 1 Introduction Chapter Two ................................................................................................ 5 Arterial Function and Structure 2.1 Historical perspectives 2.2 Large artery wall structure 2.3 Mechanical properties of arteries: evolutionary perspectives 2.4 The behaviour of the large arteries under a distending pressure 2.5 Experimental investigation of aortic wall properties 2.6 Essential non-linear and pressure-dependent properties of aortic stiffness 2.7 The relation between arterial stiffness and hypertension 2.8 Pathophysiological mechanisms associated with arterial stiffening 2.9 Animal models of arterial stiffness 2.10 Measurement of arterial stiffness Chapter Three ............................................................................................ 37 Logical Treatments of Arterial Hypertension 3.1 Nitric oxide 3.2 Angiotensin-converting enzyme inhibition 3.3 Diuretics 3.4 Angiotensin II type 1 receptor and calcium channel blockers 3.5 Other interventions 3.6 Conclusion viii Table of Contents Chapter Four .............................................................................................. 41 Common Methodology Used in the Investigation of Arterial Function and Structure 4.1 Measurements of body weight, body length and conscious tail-cuff systolic blood pressure 4.2 Haemodynamic measurements 4.3 Aortic calcium quantification 4.5 Statistical analysis 4.4 Histomorphometry Chapter Five .............................................................................................. 53 Regional Aortic Wall Stiffness: Comparison in Normotensive, Hypertensive and Aortic Calcification Rat Models 5.1 Introduction 5.2 Methods 5.3 Results 5.4 Discussion 5.5 Conclusion 5.6 Perspectives Chapter Six ................................................................................................ 71 Aortic Stiffness is Associated with Vascular Calcification and Remodelling in a Chronic Kidney Disease Rat Model 6.1 Introduction 6.2 Materials and Methods 6.3 Results 6.3 Discussion 6.4 Conclusion Chapter Seven ............................................................................................ 83 Brief Antihypertensive Treatment with ACE Inhibition (perindopril) Improves Aortic Distensibility in Spontaneously Hypertensive Rats 7.1 Introduction 7.2 Methods 7.3 Results 7.4 Discussion 7.5 Conclusion 7.6 Perspectives Vascular Function and Structure in the Rat Aorta ix Chapter Eight ........................................................................................... 101 Withdrawal of Chronic Antihypertensive Treatment with Perindopril Offsets Aortic Distensibility by a Non-pressor Mechanism in Lewis Polycystic Kidney Disease Rats 8.1 Introduction 8.2 Methods 8.3 Results 8.4 Discussion 8.5 Conclusion 8.6 Perspectives Chapter Nine ............................................................................................ 119 Conclusions Chapter Ten ............................................................................................. 125 Future Research References ............................................................................................... 127 LIST OF FIGURES Figure 2.1 An example of photomicrographs of cross sections of a WKY rat thoracic (left) and abdominal (right) aorta. Figure 2.2 Collagen (left) and elastin (right) content (mean±SD) in the ascending (A), descending (B) thoracic aorta and abdominal supraceliac (C), suprarenal (D), and midinfrarenal (E) aorta. Figure 2.3 (A) Stress–strain inflation curves for the aortae of four different species at 10 °C, showing increased slope (modulus) with increased stress. Figure 2.4 Einc versus external radius for a dog femoral artery. Figure 2.5 Einc versus MAP of the thoracic aorta (squares), abdominal aorta (triangles), femoral artery (crosses) and carotid artery (circles) Figure 2.6 The contribution of elastin and collagen fibres to the tension- radius response of human iliac arteries. Figure 2.7 Plot of the relationship between tension and expansion of an artery (A) and balloon (B) during inflation. Figure 2.8 Microscopic difference between atherosclerosis and arteriosclerosis. Figure 2.9 Osteogenesis regulation of aortic calcification. Figure 4.1 An example of a tail-cuff recording of SBP using the IITC Life Science blood pressure system Figure 4.2 In-vivo experimental set up. Figure 4.3 An example of beat-to-beat recording and calculation of transit time from second derivatives of blood pressure waveforms. Figure 4.4 Blood pressure, PWV and HR beat-to-beat signal recordings during PE and SNP infusions in one WKY rat. Figure 4.5 Flame atomic absorption spectrometer used for determination of aortic calcium content. Figure 4.6 An example of a 4µm section stained with Shitaka’s Orcein. 50 Figure 4.7 Force equilibrium in two equal halves of vessel. Table 5.1 In vivo mechanical characteristics Figure 5.1 Sample experimental recording. Figure 5.2 PWV-MAP relationship. Figure 5.3 Thoracic PWV-MAP of WKY, VDN and SHR. Figure 5.4 Abdominal PWV-MAP of WKY, VDN and SHR. xii List of Figures Figure 5.5 Calculated elastic modulus and wall stress of the aortic segments. Figure 6.1 PWV-MAP curves are averaged over 5mmHg pressure steps in Lewis (open triangles) and LPK (closed triangles) ±SEM. Figure 6.2 Aortic calcium content of LPK showed a 6.5-fold increase compared with Lewis (Lewis: 66.8±5.7 µmol·g-1; LPK: 434.0±118.3 µmol·g-1, *P<0.001). Figure 6.3 Typical histological sections stained with Shitaka’s Orcein (top) and M.S.B. (bottom), showing a longitudinal section of the thoracic descending aorta Figure 7.1 Body weight during the 5 months following ACE inhibition with perindopril at 3mg/kg/day in all strains of rats. Figure 7.2 PWV–MAP curves for all four groups of rats (n=6 in each group) in the whole (top panel), thoracic (middle panel) and abdominal (bottom panel) aorta. Figure 7.3 Typical 4µm cross sections of the thoracic aorta. Figure 7.4 MCSA (µm2) of the thoracic and abdominal aorta in untreated SHR, Figure 7.5 Total calcium levels (µmols.g-1) in the thoracic (white bar) and abdominal (black bar) aorta in 25-week-old WKY, untreated SHR, SHR(Tx)y and SHR(Tx)o rats. Figure 8.1 Effect of chronic ACE inhibition with perindopril (3mg/kg) in the Lewis (A) and LPK (B) from 6 weeks of age on SBP. Figure 8.2 Isobaric PWV-MAP curves in the Lewis (A) and LPK (A). Figure 8.3 Aortic calcium content. Figure 8.4 MCSA of the thoracic aorta in all groups of rats. Figure 8.5 Elastic modulus/wall stress ratio of the thoracic aorta in all groups of rats. LIST OF TABLES Table 5.2 Relationship between PWV and MAP of WKY, SHR, and VDN rats. Table 5.3 Morphometric parameters of the descending thoracic aorta. Table 6.1 Basal haemodynamic parameters in the Lewis and LPK. Table 6.3 Morphometric parameters of the descending thoracic aorta. Table 7.1 ACE inhibition and haemodynamic parameters from control, SHR(Tx)y, SHR(Tx)o and control SHR rats. Table 8.1 Haemodynamic parameters in anaesthetised 6-week-old, 12- week-old untreated and 12-week-old perindopril-treated LPK and Lewis rats. Table 8.2 Morphometric parameters of the descending thoracic aorta. ACKNOWLEDGEMENTS Without the help and support I received, a body of work such as this would not have been possible, and it would have been a much lesser book. This section is especially devoted to paying tribute to the myriad contributions of my advisors and collaborators, and the support of my family and friends. I want to thank my doctoral advisor, Alberto Avolio. It has been an honour to be his first PhD student at Macquarie University. He has taught me, both consciously and unconsciously, how good science is done. His theoretical insight and philosophical outlook on the topic and in life assisted me substantially in developing my research acumen. His enthusiasm is so motivational and will continue to inspire me throughout my entire scientific career. Various other individuals have also contributed to this book by providing advice and support throughout the course of my candidature. In this regard I would especially like
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