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UNIVERSITY OF CALGARY The Effect of Carbon Dioxide on Airway Tone by Tamer Y. El Mays A THESIS SUBMITTED TO THE FACULTY OF GRADUATE STUDIES IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE DEPARTMENT OF CARDIOVASCULAR/RESPIRATORY SCIENCES CALGARY, ALBERTA April, 2007 © Tamer Y. El Mays 2007 UNIVERSITY OF CALGARY FACULTY OF GRADUATE STUDIES The undersigned certify that they have read, and recommend to the Faculty of Graduate Studies for acceptance, a thesis entitled “The Effect of Carbon Dioxide on Airway Tone" submitted Tamer Y. El Mays in partial fulfilment of the requirements of the degree of Master of Science. Supervisor, Dr. Francis HY Green, Dept. of Cardiovascular/Respiratory Sciences Dr. Marc J. Poulin, Dept. of Cardiovascular/Respiratory Sciences Dr. William A. Whitelaw, Dept. of Cardiovascular/Respiratory Sciences Dr. Richard J. Wilson, Dept. of Cardiovascular/Respiratory Sciences Dr. Neil Eves, Faculty of Kinesiology Date ii Abstract The Buteyko Breathing Technique (BBT) has received interest over the last few years as a non pharmacological complementary therapy for asthma. The BBT involves breath holding in order to raise PCO2 which is believed to result in broncho-relaxation and relief of asthma. Clinical trials indicate that asthmatics using BBT need less asthma medications. However, studies of the physiological basis for the theory have not yet been undertaken. The objective of this study was to determine the effects of inhaled CO2 on constricted airways in allergic asthma as well as evaluating the effects of CO2 on bronchial smooth muscle (BSM) tone in vitro. An animal model of acute on chronic asthma was developed using Brown Norway Rats immunized and challenged with ovalbumin. A biphasic asthmatic response was observed using non-invasive whole body plethysmography. Using this model, the effects of different concentrations of CO2 on the response of allergic asthma were evaluated. We found that inhaled CO2 caused a significant (p<0.001), rapid and reversible dose-dependent relaxation of constricted airways during the late phase of the asthmatic response. Furthermore, hypocapnic challenge increased bronchial smooth muscle resting tone in vitro and significantly diminished substance P-induced epithelium-dependent relaxation of bronchial smooth muscle. The smooth muscle response to hypocapnia was pH dependent and in part dependent on L-type Ca2+ channels. Our results show that CO2 is a rapid and effective bronchial smooth muscle relaxant with potential for the treatment of acute asthma in humans. These animal studies provide supportive evidence to some aspects of Dr. Buteyko’s theory. iii Acknowledgements I would like to acknowledge my supervisor, Dr. Francis Green and thank him for being a great teacher, a guide and a friend. Without him, this work would not be possible. I owe special thanks to my supervisory committee members Drs. Marc Poulin, William Whitelaw and Richard Wilson, for their guidance and support. I would like to acknowledge my colleagues in the Respiratory Research Group at the University of Calgary for their collaboration and solidarity. In our group, special thanks to Dr. Rommy Koetzler for her support and advice. The technical assistance to Marija Susnjar, Elizabeth French, Hannah Park, Tara Stapleton Artee Karkhanis and Marnie Cudmore is greatly appreciated. I would like to express my sincere gratitude to the co-operative groups involved in this work: the lab of Drs. Morley Hollenberg and Roland Auer. My special thanks to the staff of Dr Hollenberg: Dr. Mahmoud Saifedine and Zhenguo Yu, for their contributions and help through out this project. Special thanks to my friend Dr. Abdel Aziz Shaheen for his great help in statistics. My sincere thanks for my friends Dr. Ahmed Al-Sabi’ and Fateh Zaamout for their endless support and kindness. I am especially grateful to my parents, my brothers, my grandmother, and my small family; my wife and precious children for their continuous support and love. Love you all and keep blooming in the sky... Tamer Y. El Mays February, 2007 iv Dedication I dedicate this thesis to my Parents, my brothers Hesham and Amro, and my grandmother. To my dear wife Julia and my lovely children Youssef and Inas. v Table of Contents Approval Page.................................................................................................................ii Abstract......................................................................................................................... iii Acknowledgements ........................................................................................................iv Dedication .......................................................................................................................v List of Tables................................................................................................................viii List of Figures ................................................................................................................ix List of Symbols, Abbreviations and Nomenclature ........................................................xii CHAPTER ONE: INTRODUCTION ..............................................................................1 1.1 The respiratory system...........................................................................................1 1.1.1 Airways anatomy...........................................................................................1 1.1.2 Airway smooth muscle...................................................................................2 1.1.3 The respiratory circulation .............................................................................4 1.1.4 Gas exchange.................................................................................................4 1.2 Asthma ..................................................................................................................5 1.2.1 Definition ......................................................................................................5 1.2.2 Prevalence .....................................................................................................6 1.2.3 Etiology and pathogenesis..............................................................................6 1.2.3.1 Atopic asthma.......................................................................................7 1.2.3.2 Non-atopic asthma................................................................................8 1.2.3.3 Mucus plugs .......................................................................................10 1.2.3.4 Remodelling .......................................................................................11 1.2.4 Clinical manifestation of asthma ..................................................................11 1.2.5 Breathing disorders associated with asthma..................................................11 1.2.6 Treatment of asthma ....................................................................................13 1.2.6.1 Conventional treatments .....................................................................13 1.2.6.2 Alternative treatments.........................................................................16 1.3 Hypothesis...........................................................................................................20 1.4 Objectives............................................................................................................20 CHAPTER TWO: THE EFFECT OF INHALED CARBON DIOXIDE ON CONSTRICTED AIRWAYS IN AN ANIMAL MODEL OF ASTHMA ..............21 2.1 Introduction .........................................................................................................21 2.2 Materials..............................................................................................................22 2.3 Methods...............................................................................................................25 2.3.1 Whole body plethysmography......................................................................25 2.3.2 Experimental design ....................................................................................30 2.3.2.1 Baseline pulmonary function in unchallenged rats ..............................30 2.3.2.2 Experimental protocols.......................................................................31 2.3.3 Statistical analysis........................................................................................39 2.4 Results.................................................................................................................39 2.4.1 Whole body plethysmography......................................................................39 2.4.2 Blood gas analysis .......................................................................................42 vi 2.4.3 Pulmonary Function.....................................................................................44 2.4.4 Qualitative pathologic assessment of the multiple allergen-challenged model...........................................................................................................47 2.4.5 Quantitative measurements of collagen ........................................................48 2.5 Discussion ...........................................................................................................50 CHAPTER THREE: THE EFFECTS OF CARBON DIOXIDE ON BRONCHIAL SMOOTH MUSCLE TONE .................................................................................54
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