Intolerance in Heart Failure
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1 Novel Targets to Attack Exercise Intolerance in Heart Failure - Skeletal Muscle - Volker Adams, PhD ESC, Paris 3. Aug. 211 UNIVERSITÄT LEIPZIG H E R Z Z E N T R U M Nothing to disclose
2 Myers et al. NEJM 22 Exercise Capacity and Mortality
3 Adjusted all cause mortality Exercise Capacity and Mortality 2867 men perform an exercise test (Bruce protocol or ramp test) Follow-up period 22.8 years (mean 7.8±5.1 years) 5 65 yrs (n=1 658) > 65 yrs (n=1 29) < > 8. METs < > 8. METs Nylen et al. J Am Geriatr Soc 21
4 central hemodynamic pulmonal function Exercise training in healthy and diseased skeletal muscle endothelial function
5 Targets of Exercise Training Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Oxidative stress Energy Metabolism Stem cells Capillarization Micro-RNA
6 Targets of Exercise Training Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Oxidative stress Energy Metabolism Stem cells Capillarization Micro-RNA
7 Effects of Exercise Training on Circulating Cytokine Levels Randomized study, 24 patients with CHF (NYHA II/III) and 2 controls; exercise training over a period of 12 weeks. Sampling of blood at begin and after 12 weeks. TNF-a (pg/ml) IL-6 (pg/ml) 1. p<.5 1. p< p 5. p Beg. 12 w Beg. 12 w Control CHF. Beg. 12 w Beg. 12 w Control CHF Adamopoulos et al., J Am Coll Cardiol 22
8 Local IL-6 Expression [rel. units] Local TNF-a Expression [rel. units] Control Training Local IL-1ß Expression [rel. units] Effects of Exercise Training on Local Cytokine Expression # 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 Oxidative stress Energy Metabolism Stem cells Capillarization Micro-RNA
10 Ubiquitin proteasome pathway chronic heart failure denervation immobilisation weightlessness Murf 1/ MAFbx (E3 ubiqitin ligases) proteins proteins ubiquitin ubiquitin protein complex skeletal muscle cell proteolysis
11 Study Design - Exercise Training and E3 Ligase Expression CHF group LAD Ligation sham OP sham group Exercise Training (3weeks, 2x per day, 3 min) Control Exercise Training (3weeks, 2x per day, 3 min) Control Analysis of Gene Expression
12 mrna expression [arb. Units] mrna expression [arb. Units] Exercise Training and MAFbx / Murf-1 Expression MAFbx Murf-1 3 P<.5 P<.1 4 P<.5 P< p 2 p 1 1 sham-c sham-t CHF-C CHF-T sham-c sham-t CHF-C CHF-T
13 Murf1 /18S Correlation of Murf-1 Expression and TNF-alpha 2 r=.62 P< TNF - a [ pg /mg Protein] Adams et al. Cardiovasc Res 27
14 Change in Murf-1 expression [%] Change in Murf-1 expression by exercise training - NYHA III Study D Muscle mass (%) 5 * p< r= Control p Training p D Murf-1 (%) * -2
15 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
16 Study Design and Methods WT and MuRF-KO IP injection of TNF-a (1 ng/g) IP injection of NaCl 24h inoculation time Measurement of muscle contractility (soleus, force-frequency relation) Expression analysis of MuRF1 and contractile proteins
17 Force [g/sq cm] Force Frequency Relation in Soleus Muscle WT control Murf-1 -/- control Murf-1 -/- + TNF alpha WT + TNF alpha Hz 1 Hz 25 Hz 5 Hz 75 Hz 1 Hz 125 Hz 15 Hz Frequency [Hz] Murf-1 -/- + TNF alpha WT + TNF alpha WT control Murf-1 -/- control Adams et al., J Mol Biol 28
18 Pathophysiological Model for the TNF-a Induced Reduction in Muscle Contractility Dexa NF-kB TNNT-3 TNF - a p38 MAPK Murf - 1 Translation Elongation Muscle contractility ROS Adams et al., J Mol Biol 28
19 Myostatin knock out animal Belgian blue Myostatin negative regulator of muscle mass
20 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
21 IGF-I mrna Expression [ar. Units] IGF-1 and Skeletal Muscle Mass IGF-1 transgenic animals (IGF-1 under the control of skeletal muscle promotor) Local IGF-I Expression vs. Muscle Cross-Sectional Area 15 r=.75 p= Muscle CSA [cm 2 ] Musaro et al.; Nature 21 Hambrecht et al.; J Am Coll Cardiol 22
22 [% 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
23 Targets of Exercise Training Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Oxidative stress Energy Metabolism Stem cells Capillarization Micro-RNA
24 Production and Detoxification of Reactive Oxygen Species Production Detoxification NAD(P)H oxidase xanthine oxidase myeloperoxidase uncoupled NOS Enzymes of the respiratory chain SOD O 2 - H 2 O 2 GSSG GSH GPX 2 H 2 O Fe 2+ Fe 3+ OH Catalase H 2 O + O 2
25 enzyme activity (U/mg) Lipid peroxides (µmol/mg) enzyme activity (U/mg) enzyme activity (U/mg) Effects of Exercise Training on Oxidative Stress - Radical Scavenger Enzymes - SOD 1. GPX * * Catalase. 4 HS B 6Mo CHF Training B 6Mo CHF Control p 6 HS B 6Mo CHF Training B 6Mo CHF Control 3 5 * * 2 1 * * HS B 6Mo B 6Mo CHF Training CHF Control HS B 6Mo B 6Mo CHF Training CHF Control Linke et al., Circulation 25 * p<.5 vs. HS p<.5 vs. CHF Training B
26 D ROS generation in leucocytes (%) Effects of Exercise Training on ROS Leipzig exercise intervention in CHF and ageing trail (LEICA) DHE 5 p<.1 25 ROS DHE DHE ox Healthy control Healthy training CHF control CHF training -25 DHE: Dihydroxyethidium, is able to pass the cell membrane, non-fluorescent DHE ox : Dihydroxyethidium oxidized form, is no longer able able to pass the cell membrane, fluorescent FACS -5
27 Targets of Exercise Training Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Oxidative stress Energy Metabolism Stem cells Capillarization Micro-RNA
28 Capillaries / muscle fiber Capillary Density in Skeletal muscle of CHF Rabbit model of CHF > LAD ligation Patients with CHF 1.5 p< p< Capillaries / mm Healthy CHF Healthy CHF Nusz et al., Am J Physiol 23 Williams et al., J Card Fail 24
29 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., Circ Heart Fail 21
30 Targets of Exercise Training Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Oxidative stress Energy Metabolism Stem cells Capillarization Micro-RNA
31 Definition of micro-rna mirnas are small (~ 2-3 nucleotides) non-coding RNAs which are highly conserved from plants to mammals. Their known functions are to inhibit protein translation or to enhance mrna degradation. Mammalian genomes are predicted to encode 2 to 5 unique mirnas.
32 Micro-RNAs in the Skeletal Muscle and Exercise Training Endurance Resistance (single bout) single bout Training mir-1 mir-133a mir-26 mir-23a mir-1 mir-181 mir-17 mir-696 mir-21 mir-79 mir-72 Güller and Russell., J Physiol 21
33 mir-696 expression PGC-1a expression PGC-1a expression (arb Units) Targets for Specific mirna 3 r=.64 2 sedentary group training group (4wk) 1 mir-696 PGC-1a 12 p<.5 15 p< mir-696 expression (arb Units) 8 1 Training Aerobic metabolism 4 5 mir-696 Mitochondrial content Fatty acid oxidation sedentary training sedentary training PGC-1a Fiber type shift
34 Targets of Exercise Training Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Oxidative stress Energy Metabolism Stem cells Capillarization Micro-RNA
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