Clinical trials and Myokines



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Clinical trials and Myokines Hellsten Ylva Background: Cardiovascular disease is a major cause of death in the western world. Regular physical activity is known to be highly beneficial for improving cardiovascular health status. Methods:The current abstract is based on the authors own data and a literature review. The authors own studies have all been accepted by the local ethics committee. In intervention studies the effect of physical training on cardiovascular risk parametres as well as on specific biochemical systems in the cardiovascular system have been studied. Changes in plasma myokine concentrations have been assessed in acute exercise and training interventions in different populations. Results: The conducted studies within the cardiovascular area clearly show that regular physical activity is highly beneficial for vascular function and for reducing blood pressure both in aged individuals and in individuals with life style related disease. An ongoing study at The University of Copenhagen conducted in collaboration with Martin Daumer will show to what extent estrogen loss in connection with menopause in women can be replaced by physical activity and thereby retard the progressive risk for cardiovascular disease commonly seen in women after menopause. Many studies have documented that myokines are elevated in response to exercise but more systematic investigations of how specific myokines are altered in response to various types of intensities and duration of exercise and in different subsets of the population is required to more solidly assess their usefulness as markers of physical health status. Discussion/conclusion: Based on data from our own studies and studies in the literature, physical activity should be recommended for prevention of cardiovascular disease and should be a primary recommendation in treatment of patients with cardiovascular disease to improve overall cardiovascular health. Use of myokines as markers of health status may be useful but selection of the best markers should await further evidence from systematic studies. <!--[endif]--> PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Clinical trials and Myokines The study of cardiovascular function and health and the impact of physical activity Can myokines be used as indicators of training/health status? Ylva Hellsten, Department of Nutrition, Exercise and Sports, Copenhagen University, Denmark PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Cardiovascular function and growth- why are we interested? Sports performance The limits of endurance performance lies in the cardiovascular system Health-hypertension 8 million people die every year in the world on as a consequence of elevated blood pressure Approximately 25-40% of the adult Danish population have elevated blood pressure Relation to physical activity: For every 2 mm of reduction in blood pressure it is estimated that the risk of death by stroke is reduced by 10 % and the risk of death by ischemic cardiac disease PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014 reduced by 7%

Aspects of the cardiovascular system that we study in man Red blood cells Heart Platelets Oxygen Arteries Arterioles Veins Venules A heart pumps out > 200 million liters of blood in a lifetime Capillaries PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014 Microcirculation

How we study blood flow regulation/vascular function in humans Infusion of compounds Blood sampling Knee extensor model Catheterization Blood flow measurements Microdialysis Pharmacological interventions Muscle biopsy Rodents Microscopy Cell culture PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Muscle blood flow in relation to workload: a precise balance between metabolic demand and oxygen delivery Leg blood flow Exercise intensity Saltin et al. 1998 PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

In aging and cardiovascular disease vascular function in the arm is impaired Vasodilation in arm Normotensive Hypertensive Taddei, S. et al. Hypertension 1997;29:736-743 PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

In aging vascular function in the leg is impaired Femoral arterial acetylcholine infusion Infusion of Ach Blood sampling Young Aged Tr Aged Untr Nyberg et al. 2012 VO 2 max (ml/min/kg bw) Young: 45±2 Aged utr: 26 ±1 Aged Tr: 46± 3 PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Can you improve cardiovascular health by training in aged individuals- or is it too late? Training of 27 healthy men 60-65 years of age Spinning 2 x week + crossfit 1 x week for 8 weeks Resveratrol supplementation PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Eight weeks of physical activity increases maximal oxygen uptake in aged sedentary men ~19% ~14% PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Eight weeks of physical activity lowers mean arterial blood pressure in aged sedentary men And a number of other parametres related to cardiovascular health were improved by training PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Department of Nutrition, Exercise and Sports Conclusive remark Training is highly effective in improving cardiovascular health, including in aging and cardiovascular disease (but be careful with your supplements) PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Copenhagen Women Study Women s health during the menopausal transition (45-55 years) PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Department of Nutrition, Exercise and Sports Background for study Due to the effects of estrogen, women have a lower risk than men for cardiovascular disease before menopause After menopause the loss of estrogen -increases the risk of cardiovascular disease -increases loss of bone mass -increases loss of muscle mass -increases the risk of metabolic disease Exercise affects similar molecular pathways as estrogen and has marked beneficial effects on health PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Department of Nutrition, Exercise and Sports Hypotheses of study Exercise training will limit the adverse effects of estrogen loss on: -Vascular function and growth -Cardiac structure and function -Metabolic status and fat distribution -Bone density -Muscle mass And will improve: -the level of daily activity -reduce depression and anxiety Motivational coaching will have positive effects on adherence to physical activity. PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Collaborative partners outside department: Henrik Larsson, Glostrup Hospital, MR scanning Charlotte Suetta, Glostrup Hospital, DXA scanning Martin Daumer, The Human Motion Institute, Munich, Assessments of daily activities Niklas Rye Jørgensen, Glostrup Hospital, bone metabolism, Ruth Frikke Schmidt, Rigshospitalet, Genetics Lars Bo Nielsen, Rigshospitalet Cardiovascular markers Lotte Enevoldsen, Rigshospitalet, Fat and glucose metabolism PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Overall experimental design Post-menopausal Spinning- Training N=30 Healthy women 45-55 yrs Control N=30 Pre-menopausal Spinning- Training N=30 Intervention for 10 months Testing pre and at 3 and 10 months Control N=30 N=15+15: Fat metabolism/insulin sensitivity N=15+15 Vascular and cardiac function PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Measured parametres in study CHARACTERIZATION OF SUBJECTS VO 2 max Body composition incl bone density Plasma lipid status Oral glucose tolerance test Food registration Questionnaires (Social and psychological aspects) Muscle and fat biopsies SPECIFIC ASSESSMENTS Assessment of vascular function (with focus on prostaglandins) Assessment of cardiac structure and function (MR and echo doppler) Assessment of angiogenic potential Assessment of daily activities Assessment of insulin sensitivity by euglycemic hyperinsulemic clamp Assessment of fat metabolism Interviews (Sociology) Coaching (Psychology) PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Department of Nutrition, Exercise and Sports Could we use myokines or other compounds for motivational feedback and as markers for training and health status? PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Department of Nutrition, Exercise and Sports What factors do we associate with health improvements? Decreased blood pressure Improved vascular function Improved cardiac function Improved muscle function Improved fitness level (maximal oxygen uptake) Improved insulin sensitivity (Decreased BMI) (Increased muscle/fat ratio) How can we assess these parameters? Or even better, can we find a single representative marker for these parameters? Myokines? PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

What are myokines? Pedersen B K J Exp Biol 2011;214:337-346 PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Department of Nutrition, Exercise and Sports Sted og dato PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Department of Nutrition, Exercise and Sports Qualities of a useful exercise/training marker: Similar behaviour in different people Not too variable by factors other than exercise Sufficiently large change in plasma with exercise/training Reliable time course Easy to conduct reliable measurements Should be a motivational marker PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Myokines that increase in plasma after exercise Department of Nutrition, Exercise and Sports Raschke & Eckel, Mediators of Inflammation, 2013 PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Department of Nutrition, Exercise and Sports Plasma myokine/cytokine response to strenuous endurance exercise Figure 1. The plasma cytokine response to strenuous exercise. FEBBRAIO M A, and PEDERSEN B K FASEB J 2002;16:1335-1347 Copyright Federation of American Societies for Experimental Biology (FASEB). Please see the journal's copyright policies at www.fasebj.org to obtain permission to use this image. PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Serum levels of IL-6 after 30 min of concentric or eccentric exercise Conc Ecc During After Proposed roles of IL-6 in exercise Glycogen censor Fat oxidation Lipolysis TNF-a suppressor Brunsgaard et al. J.Physiol. 1997 PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

IL-6 transcription in response to exercise-relationship with glycogen levels Figure 2. Biphasic transcription of IL-6 in isolated nuclei from muscle biopsies obtained before and during exercise. Department of Nutrition, Exercise and Sports FEBBRAIO M A, and PEDERSEN B K FASEB J 2002;16:1335-1347 PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Plasma IL-6 level in response to 3 h of exercise is similar before and after training Pre training Post training Fischer et al. Am J Physiol. 2004 PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Department of Nutrition, Exercise and Sports Plasma Irisin in response to 45 min of exercise- effect of training Proposed role of Irisin Browning of fat (leading to increased energy expenditure through dissipation of heat) Norheim et al. FASEB J, 2014 PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Department of Nutrition, Exercise and Sports Plasma IL-15 in response to 3 h of cycling exercise Muscle IL-15 mrna in response to resistance exercise Nielsen et al. JPhysiol 2007 Proposed roles of IL-15 in exercise Increased hypertrophy of muscle Decrease in fat mass PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Department of Nutrition, Exercise and Sports Plasma levels of adiponectin, leptin, IL-6 and CRP before and after training in lean and obese persons Proposed roles of adiponectin Insulin resistance Protection of endothelial dysfunction Adiponectin Leptin Proposed roles of Leptin Reduces hunger Increases energy expeniditure IL-6 CRP Proposed role of CRP Pro-inflammatory Samjoo et al. 2013 PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014

Department of Nutrition, Exercise and Sports Parametres associated with health improvements Decreased blood pressure Improved vascular function Improved cardiac function Improved muscle function Improved fitness level (maximal oxygen uptake) (Decreased BMI) Improved insulin sensitivity (Increased muscle/fat ratio) Potential indicators Fasting blood glucose levels (Glucose) Plasma HDL/LDL ratio (HDL/LDL levels) Plasma markers of inflammation (Adipokines: leptin, TNFa, PAI-1, Resisitin) Plasma markers of atherosclerosis (CRP, VCAM-1, ICAM-1) Markers of vascular function (Endothelin-1, follistatin related protein-1) Resting heart rate Resting blood pressure Can any one of myokines or other markers be used? PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014 Time trial (walking, running)

Acknowledgement Group and collaborators Bengt Saltin Stefan Mortensen Michael Nyberg Jens Bangsbo Lasse Jensen Ane Hansen Karina Olsen Gemma Kroos Jens Jung Nielsen Ulrik Frandsen Jan Lynge Funding Danish Council for Independent Research-Medical Sciences Lundbeck Foundation Novo Nordisk Foundation PeerJ http://dx.doi.org/10.7287/peerj.preprints.367v1 CC-BY 4.0 Open Access received: 15 Apr 2014, published: 15 Apr 2014