KININS AND KININ RECEPTORS IN THE PATHOGENESIS OF HEART FAILURE Antti Kuoppala Doctoral thesis University of Helsinki Wihuri Research Institute, Helsinki, Finland Academic Dissertation To be presented by permission of the Medical faculty of the University of Helsinki, for public examination in auditorium 4 at Meilahti Hospital on December 5th 2002 at 12 noon. The supervisors of the thesis Doc Ken Lindstedt Jorma Kokkonen, Ph.D. Communicated by Prof Olli Vuolteenaho Doc Ilkka Pörsti ISBN 952-91-5388-0 (paperback) ALSO AVAILABLE IN ELECTRONIC FORMAT ISBN 952-10-0834-2 (PDF) URL:http://ethesis.helsinki.fi/ Yliopistopaino Helsinki 2002 To my Lord Page CONTENTS .......................................................................................................................... 5 ABBREVIATIONS ............................................................................................................... 9 LIST OF ORIGINAL PUBLICATIONS ............................................................................ 11 REVIEW OF THE LITERATURE .................................................................................... 12 1. Heart Failure .................................................................................................................. 12 1.1. Pathogenesis of heart failure ...................................................................................... 12 2. The kininogen-kallikrein-kinin system ........................................................................ 15 2.1. Bradykinin and kallidin .............................................................................................. 15 2.2. Regulation of kinin concentration .............................................................................. 16 2.2.1. Formation of kinins from kininogens ................................................................... 16 2.2.1.1. Kallikreins ...................................................................................................... 16 2.2.2. Degradation of kinins ........................................................................................... 19 2.3. BK Receptors ............................................................................................................. 20 2.3.1. BK-2R .................................................................................................................. 21 2.3.1.1. Physiological effects of BK-2R ...................................................................... 21 2.3.1.2. The regulation of BK-2R ................................................................................ 22 2.3.1.3. Intracellular cascades and second messengers of BK-2R ............................... 25 2.3.2. BK-1R .................................................................................................................. 29 2.3.2.1. Physiological effects of the BK-1R ................................................................ 30 2.3.2.2. Regulation of BK-1R ...................................................................................... 30 2.3.2.3. Second messengers of BK-1R ........................................................................ 32 3. Influence of kinins on diseases contributing to the pathogenesis of heart failure and on heart failure directly .......................................................................................... 33 3.1. Diabetes mellitus ......................................................................................................... 33 3.1.1. Effect of diabetes in the control of expression of kininogen-kallikrein-kinin system components .............................................................................................. 33 3.1.2. The kininogen-kallikrein-kinin system in the pathophysiology of diabetes: its relation to insulin and glucose metabolism .......................................................... 34 3.1.3. Bradykinin and vasodilatation in diabetes ............................................................ 35 3.2. Hypertension ............................................................................................................. 35 3.2.1. Regulation of the kininogen-kallikrein-kinin system in hypertension ................. 35 3.2.2. The systemic vascular effects of the kininogen-kallikrein-kinin system .............. 36 3.2.3. The renal kininogen-kallikrein-kinin system in hypertension .............................. 38 3.3. The role of kinins in ischemic and ischemia-reperfusion injuries .............................. 39 3.3.1. Ischemia-reperfusion injury .................................................................................. 39 5 3.3.2. Preconditioning .................................................................................................... 41 3.4. The kininogen-kallikrein-kinin system in the pathogenesis of heart failure .............. 44 3.4.1. The components of the kininogen-kallikrein-kinin system in heart failure ......... 44 3.4.2. Kinins and endothelial dysfunction in heart failure ............................................. 45 3.4.3. The effect of kinins on myocardial oxygen metabolism and cardiac function in heart failure .................................................................................................... 46 3.4.4. Kinins in left ventricular hypertrophy .................................................................. 47 AIMS OF THE PRESENT STUDY .................................................................................... 50 MATERIALS AND METHODS ......................................................................................... 51 1. Kinin degradation (I, II) ................................................................................................ 51 1.1. Acquiring human heart samples (I, III) ...................................................................... 51 1.2. Preparation of human cardiac membranes for enzymologic measurements (I) ......... 51 1.3. Preparation of human plasma (II) ............................................................................... 51 1.4. Determination of kinin degradation (I, II) .................................................................. 51 1.5. Determination of ACE activity (I) ............................................................................. 52 1.6. Reverse-phase high-performance liquid chromatography analysis (I, II) .................. 52 1.7. N-terminal sequence analysis of kinin peptides (I, II) ............................................... 53 2. Kinin receptors (III, IV) ................................................................................................ 53 2.1. Experimental animal preparation (IV) ....................................................................... 53 2.2. Aortic banding in SD rats (IV) ................................................................................... 53 2.3. Angiotensin II infusion in SD rats (IV) ...................................................................... 53 2.4. Echocardiography in rats (IV) .................................................................................... 54 2.5. Detection of BK-2R and BK-1R mRNA by competitive RT-PCR (III, IV) .............. 54 2.6. Detection of BK-2Rs and BK-1Rs by Western blotting (III, IV) .............................. 55 2.7. Histo- and immunohistochemical staining of normal and failing hearts (III, IV) ..... 55 3. Statistical analysis (I-IV) ............................................................................................... 56 RESULTS .............................................................................................................................. 57 1. Degradation of kinins (I, II) .......................................................................................... 57 1.1. Human heart tissue (I) ................................................................................................ 57 1.1.1. Degradation of kallidin by human cardiac membranes (I) ................................... 57 1.1.2. Degradation of bradykinin by human cardiac membranes (I) .............................. 57 1.1.3. Comparison normal and failing hearts in their ability to degrade KD and BK (I) 57 1.1.4. Inhibition of kallidin and bradykinin degradation by enzyme inhibitors (I)......... 57 1.1.5. The role of ACE in heart tissue bradykinin metabolism (I) ................................. 58 1.2. Human plasma (II) ..................................................................................................... 58 1.2.1. Degradation of bradykinin by human plasma (II) ................................................ 58 1.2.2. Inhibition of bradykinin degradation by enzyme inhibitors (II) .......................... 59 1.2.3. The role of CPN in bradykinin metabolism (II) ................................................... 59 2. The myocardial expression of BK receptors in heart failure (III, IV) ...................... 60 6 2.1. BK receptors in human hearts (III) ............................................................................ 60 2.1.1. Expression of BK-2R mRNA in normal and failing human hearts (III) .............. 60 2.1.2. BK-2R expression and age in human hearts (III) ................................................. 60 2.1.3. BK-2R protein in normal and failing human hearts (III) ..................................... 60 2.1.4. The localization and expression pattern of BK-2Rs in diseased and normal hearts (III) ..................................................................................................................