How does Exercise Work at the Cellular/Molecular Level

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1 How does Exercise Work at the Cellular/Molecular Level Volker Adams, PhD ESC, Paris 28. Aug. 211 UNIVERSITÄT LEIPZIG H E R Z Z E N T R U M Nothing to disclose

2 Survival Exercise Training in Patients With Heart Failure - Metaanalysis - cumulative two year survival Piepoli et al. BMJ 24

3 Possible Targets for Exercise Training Myocardial function Pulmonal function Diaphragm Exercise training in healthy and diseased Skeletal muscle Endothelial function

4 Possible Targets for Exercise Training Myocardial function Pulmonal function Diaphragm Exercise training in healthy and diseased Skeletal muscle Endothelial function

5 Targets of Exercise Training Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Fiber type distribution Oxidative stress Energy Metabolism Stem cells Capillarization

6 Targets of Exercise Training Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Fiber type distribution Oxidative stress Energy Metabolism Stem cells Capillarization

7 Effects of Exercise Training on Local Cytokine Levels - animal experiments MI rats- IL-1 concentration (pg/µg protein) TNF-a concentration (pg/µg protein) CHF group LAD Ligation Exercise Training (8 weeks, 6 min/day, 55-65% VO2max) After 4 weeks Batista et al., Cytokine 21 sham OP sham group Sedentary Analysis of TNF-a and IL1 expression in soleus and EDL sedentary p<.5 sham group training sedentary training p<.5 CHI group sedentary training sedentary training sham group p<.5 p<.5 CHI group Inflammatory cytokine Anti-Inflammatory cytokine

8 Effects of Exercise Training on Local Cytokine Expression - Patients with Chronic Heart Failure- Local IL-6 Expression [rel. units] Local TNF-a Expression [rel. units] Control Training Local IL-1ß Expression [rel. units] # 3 2 * 1 Begin 6 Months Begin 6 Months $ * p<.5 versus Control $ p<.5 versus Begin # p<.5 for D versus Control Begin Gielen S et al., J Am Coll Cardiol 23 6 Months

9 Targets of Exercise Training Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Fiber type distribution Oxidative stress Energy Metabolism Stem cells Capillarization

10 Murf-1 / 18SrRNA Change in Murf-1 expression by exercise training Leipzig exercise intervention in CHF and ageing trail (LEICA) Murf-1 expression % change in Murf-1 expression 3 p<.5 5 p< p Healthy control Healthy p training CHF control CHF training -25 Healthy CHF -5

11 [% positive tissue area] Effects of Exercise Training on Local IGF-I Expression in CHF.3 IGF-I mrna Expression * p <.1 vs. Control * IGF-I Protein Content * p <.5 vs. Control 1.5 * Begin 6 months Begin 6 months Training Control Hambrecht et al.; Eur J Cardiovasc Prev Rehab 25

12 Myostatin protein expression (arb.units) Exercise Training and Myostatin Expression LAD Ligation sham OP 3 p<.5 p<.5 CHF group sham group 2 Exercise Training (4weeks, 2x/day, 3 min) Control 1 Analysis of myostatin protein expression sham CHF CHF sedentary CHF training Lenk et al., Eur J Heart Fail 29

13 Targets of Exercise Training Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Fiber type distribution Oxidative stress Energy Metabolism Stem cells Capillarization

14 Capaillaries / muscle fibre Impact of Exercise Training on Capillary Density in the Skeletal Muscle in Patients With CHF NYHA III Study D Capillary Density (%) 25 2 p<.5 2 p<.5 vwf staining Training Control -5-1 Beg. 12 w. Beg. 12 w. Training Control -15 Erbs et al., Circulation Heart Failure 21

15 Targets of Exercise Training Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Oxidative stress Energy Metabolism Stem cells Capillarization

16 Possible Targets for Exercise Training Myocardial function Pulmonal function Diaphragm Exercise training in healthy and diseased Skeletal muscle Endothelial function

17 FBF (ml/1ml tissue/min) Impact of Exercise Training on Endothelial Function Healthy subjects Stable CAD Patients Before Training After Training p<.1 Baseline D Coronary Blood Flow [%] Training group Acetylcholine 7,2 µg/min Begin 4 weeks * Acetylcholine (µg/1ml tissue/min) * p<.5 vs.control DeSouza et al., Circulation 2 Hambrecht et al., NEJM 2

18 Correction of Endothelial Dysfunction by Increasing NO Bioavailability Möbius-Winkler, Adams et al., Antioxid Redox Signal 211

19 Protein Expression [rel. units] Impact of Exercise Training on NO and ROS enos-phosphorylation (Ser1177) K K T T.1 ROS-Generation p<.1 * ROS generation [DOD 55 / min] p<.5 *. Control Training. Control Training Hambrecht et al., Circulation 23 Adams et al., Circulation 25

20 HDL

21 HDL Stimulates Vasodilation via enos Activation - Healthy vs. Diabetic Patients - Setup: Organ bath mouse aorta Phenylephrine mediated constriction to 7% max (KCl) Dilatation with increasing concentrations of HDL Sorrentino et al., Circulation 21

22 enos Ser 1177 phosphorylation (x-fold vs. unstimulated) HDL Stimulates enos Phosphorylation Healthy vs. NYHA III HDL (5 µg/ml) enos Ser 1177 phospho. enos 1. p=ns p= p< Healthy NYHA-II NYHA-III Beg NYHA-III 3 Mo Train Adams et al., unpublished

23 Stimulation of NO Production (% change vs. Baseline) HDL stimulates NO production Healthy vs. NYHA III and exercise training Incubation of human aortic endothelial cells with HDL for 1h at 37 C Quantification of NO in cell culture supernatant via ESR 4 p<.5 p< Healthy NYHA III Begin NYHA III 3 Mo Training Adams et al., unpublished

24 Progenitor Cells and Endothelial Repair - Hypthetical Working Model - RPC Resident progenitor cells Bone marrow EPC Endothelial progenitor cells EPC EPC EPC EPC EPC Growth and differentiation factors EPC EPC EPC EPC mature EC mature EC EPC derived EC RPC derived EC mature EC mature EC mature EC derived EC RPC derived EC RPC RPC RPC Smooth muscle cells RPC RPC RPC Lenk, Adams et al., J Appl Physiol 211

25 CD34 pos /KDR pos cells / 1 6 events Impact of Exercise Training in CHF on EPCs - Concentration and Function - EPC Concentration Migratory Capacity 3 p<.5 p< Beg 8 Weeks 8 Weeks detraining healthy Beg 6Mo Beg 6Mo control Training Sarto et al., J Cardiac Fail 27 Van Craenenbroek et al., Basic Res Cardiol 21

26 Possible Targets for Exercise Training Myocardial function Pulmonal function Diaphragm Exercise training in healthy and diseased Skeletal muscle Endothelial function

27 Maximal isometric force (N/cm 2 ) Impact of Chronic Heart Failure on Diaphragm Force Generation Power (Watt/m 2 ) Maximal isometric force and power generation of diaphragm bundles after LAD-Ligation compared to sham operated animals 2 p< sham CHF Sham CHF Force (%Po) van Hees HWH et al, Int. J. Cardiol. 28

28 TNF-a (pg/ml) Chronic Heart Failure and Inflammation Data from the SOLVED Trail * p<.5 vs. control * 4 * 2 control I II III NYHA functional class Torre-Amione et al, JACC 1996

29 Study Design Two months old, female C57Bl6 mice (n=4) Exercise on a treadmill 1 h per day, five times a week for four weeks (n=2) Sedentary control group (n=2) Injection of TNF-α (n=1) Injection of NaCl (n=1) Injection of TNF-α (n=1) Injection of NaCl (n=1) Force and Power analysis in diaphragm bundles, further molecular analysis

30 specific tetanic force [N/cm²] Impact of Exercise Training on TNF-a mediate Force Reduction in the Diaphragma Sedentary + NaCl Training + TNF Training + NaCl 7.5 * * * * Sedentary + TNF 5. * 2.5. * * p<.5 vs. Sedentary + NaCl and Training + TNF Stimulation frequency [Hz] Mangner et al, unpublished

31 Carbonylated Protein as Sign of Oxidative Stress 3 carbonylated proteins/gapdh [arb. units] Carbonylated Proteins (arb. Units) p< Sedentary NaCl Exercise TNF alpha Mangner et al, unpublished

32 Correlation Carbonylated Proteins and Force Generation 2 peak tetanic force [N/cm²] r = -.62 p < carbonylated proteins/gapdh [arb. units] Mangner et al, unpublished

33 Detection of Specifically Carbonylated Proteins a-actin 2-D-Alexa 488 Hydroxylamine 3 pi 1 α-actin 2 MW [kda] NaCl TNFα NaCl TNFα Control Training creatin kinase creatin kinase 2 NaCl Control TNFα NaCl Training TNFα

34 Molecular Effects of Exercise Training Heat shock proteins Opening PTP Uncoupling protein 2 Better Ca 2+ handling SERCA expression Exercise training in healthy and diseased Radical scavenger Carbonylated protein Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Oxidative stress Energy Metabolism Capillarization NO-production enos phosphorylation enos activity ROS production NADPH oxidase Radical scavenger enzymes

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