Leukocyte-derived matrix metalloproteinase-9 in patients with coronary artery disease. Associations with psychological stress and glucocorticoid sensitivity

........................................................................................................ 2 Populärvetenskaplig sammanfattning ............................................................ 3 Abbreviations ................................................................................................ 4 Introduction ................................................................................................... 5 Coronary artery disease .................................................................................. 5 Atherosclerosis ............................................................................................... 6 Leukocytes in atherosclerosis ......................................................................... 8 Peripheral blood mononuclear cells ............................................................ 8 Neutrophils ............................................................................................... 10 Matrix metalloproteinases and their endogenous inhibitors ........................ 11 MMPs and TIMPs in atherosclerosis findings from clinical studies .......... 12 Hypothalamus-pituitary-adrenal axis ............................................................ 13 Glucocorticoids and glucocorticoid receptors ........................................... 14 Glucocorticoid sensitivity .......................................................................... 16 Psychological stress and its relation to inflammation and HPA axis .............. 17 Chronic stress ............................................................................................ 17 Acute stress ............................................................................................... 18 Dysregulation of HPA axis activity in CAD patients .................................... 18 Telomeres and stress ................................................................................ 19 Aims ............................................................................................................ 21 General aims ................................................................................................ 21 Specific aims ................................................................................................. 21 Methodological considerations .................................................................... 22 Study populations ......................................................................................... 22 Isolation of PBMCs and neutrophils .............................................................. 23 Stimulation of PBMCs and neutrophils ......................................................... 24 ELISA and Luminex ........................................................................................ 24 Real time quantitative polymerase chain reaction (PCR) .............................. 25 RNA isolation ............................................................................................. 26 Quantitative PCR ....................................................................................... 26 Calculation of relative quantification values.............................................. 27 Flow cytometry ............................................................................................. 27 Gelatin zymography ...................................................................................... 28 Carotid B-mode ultrasound .......................................................................... 28 Psychological factors .................................................................................... 29 Psychological stress test ............................................................................... 30 Cortisol analysis ............................................................................................ 31 Telomere length measurement assay ........................................................... 31 Ethics statement ........................................................................................... 32 Statistics ....................................................................................................... 32 Results and Discussion ................................................................................. 33 Neutrophils as a source of MMPs and TIMPs ................................................ 33 Blood mononuclear cells as a source of MMPs and TIMPs ........................... 35 Serum and plasma levels of MMPs and TIMPs.............................................. 39 Exploring the potential associations between PBMC-derived levels of MMP9, TIMP-1 and TIMP-2 and psychological factors .......................................... 41 PBMC-derived levels of MMP-9, TIMP-1 and TIMP-2 and glucocorticoid sensitivity ..................................................................................................... 44 Release of MMP-9 and cortisol in response to acute mental stress .............. 48 The susceptibilty to stress-induced release of MMP-9 and its association with leukocyte TL and carotid atherosclerotic burden .......................................... 52 Concluding remarks ..................................................................................... 55 Acknowledgements ..................................................................................... 56 References ................................................................................................... 59 Appendix (papers I-IV) ................................................................................. 73 Original publications