Effect of Resistance Exercise on Myokine Expression in ... Song_Effect... · Kelly, Science, 2012 . Mitochondria biogenesis UCP1 activation . Metabolic disease (Obesity) Comparing

Effect of Resistance Exercise on Myokine Expression in T2DM and

Aging Model

Wook Song, Ph.D.

Health and Exercise Science Laboratory Institute of Sports Science, Seoul National University

What is the purpose of resistance training ?

Skeletal muscle : plasticity

Resistance training for T2DM and Aging?

Inactivity and Exercise

Loss of Muscle Mass and Abdominal Adiposity

Macrophage infiltration of visceral adipose tissue

Physical Inactivity

Chronic Disease: Type 2 Diabetes Mellitus, Cardiovascular Diseases, Cancer, Osteoporosis

Insulin resistance, Atherosclerosis, Tumor Growth, impaired Bone Formation

Chronic Systemic Inflammation (IL-6, TNF-α, IL-1β.)

Exercise

Physiological Changes

Beneficial Effects Of Exercise ?

(Pederson and Febbraio, 2012)

The Benefits of Resistance Exercise

- The maintenance of basal metabolic rate

- Increasing muscular strength, endurance, and mass

- Promoting independence, and helps to prevent falls in the elderly

- Improving the function of most cardiac, frail, and elderly patients, who benefits sub substantially from both upper – and lower- body exercise

Michel L. Pollock et al., AHA Science Advisory (2000)

Theoretical relation between musculoskeletal fitness and independent living across a person’s lifespan

As a person ages, his or her musculoskeletal fitness (i.e., muscular strength, muscular endurance, muscular power or flexibility) declines, such that a small impairment may result in disability. Many elderly people currently live near or below the functional threshold for dependence. High levels of (or improvements in) musculoskeletal fitness will enhance the capacity to meet the demands of everyday life and allow a person to maintain functional independence for a greater period.

Darren E. R. Warburton et al., CMAJ (2006)

How the beneficial effects of exercise can be delivered to the target tissues? Or systemically?

How can we explain the exercise effects between tissues, i.e., crosstalk between adipose tissue and skeletal muscle?

Is there any potential mediators?

Questions What I had… as an exercise/muscle physiologist…

Hypothesis

Myokine: a potential mediator of exercise

Carmen F.L et al. physiology (2013)

• Sepsis is a serious medical condition that is characterized by a whole-body inflammatory state (called a systemic inflammatory response syndrome or SIRS)

• With the discovery that exercise provokes an increase in a number of cytokines, a possible link between skeletal muscle contractile activity and immune changes was established.

Myocyte + Cytokine = Myokine

They suggest that cytokines and other peptides that are produced, expressed, and released by muscle fibers and exert either paracrine or endocrine effects should be classified as “myokines”.

Contracting muscle fibers produce and release IL-6, which induces several metabolic effect.

J Appl Physiol 98: 1154-1162, 2005

Interaction between skeletal muscle and various organs

Pedersen, B. K. & Febbraio, M. A. Nat. Rev. Endocrinol (2012)

Myokine Metabolites Exercise Metabolic Demand of Muscle Other ? Muscle-to-Nerves Interaction

Metabolic Homeostasis Age-related Mortality and Lifespan

Aging and age-related diseases in non-muscle tissue Organism’s response to oxidative stress

Organism’s response to dietary restriction

Role of Myokines During Exercise

Potential mechanism for prevention of sarcopenia or aging related diseases

Integr Med Res, Review (BH So, HJ Kim, W Song et al., 2014)

IL-15: Interleukin-15 LIF: Leukemia inhibitory factor BDNF: Brain-derived neurotrophic factor FGF-21: Fibroblast growth factor 21 SPARC: Secreted protein acidic and rich in cysteine

Fat browning Muscle-fat crosstalk

Muscle hypertrophy

Metabolic diseases

Where we are and where do we go form here?

Exerc Sport Sci Rev. 38(2) : 42-50, 2010

Some key themes driving the exercise science field in the future

Exercise mimetics □

Reactive oxygen species □

Genomics □

The muscle from inside and out □

The processing of substrate fuels □

Muscle fiber, connective tissue, bone and satellite cell integration □

Discovering biomarkers for predicting exercise and altered health settings □

Mechanical sensors and signaling regulating that control muscle size □

The link between exercising muscle and brain plasticity □

Mechanisms regulating aging and exercise induced longevity □

The role of myokines, cytokines, and adipokines are thought to impact both organ systems and organism homeostasis; the new emphasis should focus on mechanisms driving such synergism.

Exerc Sport Sci Rev. 38(2) : 42-50, 2010

Introduction of myokine

IL-15 Muscle–fat cross-talk

• IL-15 has been identified as an anabolic factor, which is highly expressed in skeletal muscle.

• Furthermore, IL-15 has been suggested to play a role in muscle-adipose tissue interaction.

Interplay between muscle and adipocyte

IL-15

Interleukin-15

IL-15 : interorgan metabolic targets. UCPs, uncoupling protine; LPL, lipoprotein lipase. Drug Discov Today 14(4) (2009)

Normal animal model (SD rat)

Metabolic disease (Type 2 diabetes model)

Evaluating the effect of acute resistance training on time-dependant change of IL-15 expression

Comparing the effect of acute (1 bout) and chronic (8 weeks) exercise on IL-15 expression

Evaluating the effect of resistance exercise on IL-15 expression in various skeletal muscle tissues (SOL. GAS, TA, EDL, etc.) of Zucker rats

Experimental results in SNU (IL-15)

The effect of endurance exercise training on glucose tolerance and IL-15 expression in skeletal muscles using type 2 diabetic rats


Purpose

- To Investigate the effects of acute and chronic resistance exercise training (8 weeks) on IL-15 expression in rat skeletal muscle.

Results - Acute bout of resistance exercise induced increase of IL-15 concentration in skeletal muscle. - Increased IL-15 expression in soleus muscle after 8 weeks of chronic resistance training where muscle growth and hypertrophy can be generally induced.


Condition Reps Set Load Rest Duration Frequ-enc

y Period

Adaption period

5 1 Weight bearing 1min 10 min 3days/wk 1 wk

ARE 10 1 50% of BW, every trial

+ 15 % of BW 2min 30 min

One session

One session

CRT 10 1 50% of BW, every trial

+ 15 % of BW 2 min 30 min 3 days/wk 8 wks

Thirty-two 10 weeks old Male SD rats

Sixteen 10 weeks old SD rats Sixteen 10 weeks old SD rats

Acute Resistance exercise(n=16) Control (n=8) 8 Weeks of Chronic Resistance Exercise (n=8)

Tissue Collection At 0h, 1h, 4h, 24h (n=4 per time point)

Tissue Collection After 48 h

Protein analysis ELISA


Fig. Time course changes of IL-15 levels following acute resistance exercise

The rats were exercised to climb a 1-m vertical (85 degree incline) ladder with weights secured to their tail. They climbed the ladder 10 times progressively. Muscle samples were obtained at resting, immediately after (0 h), 1 h, 6 h, 24 h after exercise in SOL (Fig. 1A) and TA (Fig. 1B). Data were presented as means ± S.E.M. Acute control group (Resting, n=4), acute resistance exercise groups (0 h, 1 h, 6 h, 24 h, n=4 per group). Statistical significance was set at p<0.05.

IJASS, 25:(2), 85-90, 2013


Fig. Change of IL-15 levels after 8 weeks of resistance exercise training

The rats were exercised to climb a 1-m vertical (85 degree incline) ladder with weights secured to their tail. They climbed the ladder 10 times progressively. Muscle samples were obtained at resting, immediately after (0 h), 1 h, 6 h, 24 h after exercise in SOL (Fig. 2A) and TA (Fig. 2B). Data were presented as means ± S.E.M. Acute control group (Resting, n=4), acute resistance exercise groups (0 h, 1 h, 6 h, 24 h, n=4 per group). Statistical significance was set at p<0.05.

IJASS, 25:(2), 85-90, 2013

Purpose

- To investigate the effect of endurance exercise training on glucose tolerance and IL-15 expression in skeletal muscles using type 2 diabetic rats

Results - After 12 weeks of treadmill training, reduction of body weight was observed in ZDF-Ex compared to ZDF-Con rats. - Glucose tolerance using IPGTT in diabetic rats was significantly improved in ZDF-Ex rats. - The Expression of IL-15 was significantly increased only in the SOL of ZDF-Ex rats compared to ZDF-Con

Metabolic disease model (type 2 diabetes model)

Adp.

(1 wk)

Exercise Period

1 wk 2 wk 3 wk 4 wk 5 wk 6 wk 7 wk 8 wk 9 wk 10 wk 11 wk 12 wk

duration 60 min

Load 15 m/min + 2 m/min for every 2 weeks 20 m/min

Requency 5 day/week

Twenty-four8 weeks old Zucker rats

Lean Control (ZLC-Con, n=8)

Tissue Collection After 48 h of Last intervention


Diabetic Control (ZDF-Con, n=8) Diabetic Exercise (ZDF-Ex, n=8)

12 Weeks of Resistance Exercise (n=8)


Diabetes Metab J 2013;37:358-364

Fig. Effect of 12 weeks of (A) treadmill exercise on bodyweight and (B) fasting blood glucose concentration

Values are mean±standard error of mean for n=8 in each group. ZLC-Con, sedentary Zucker diabetic fatty lean control; ZDF-Con, sedentary Zucker diabetic fatty; ZDF-Ex, exercised Zucker diabetic fatty.

aP<0.05 compared with ZLC-Con group, bP<0.05 compared with ZDF-Con group.



Fig. Effect of 12 weeks of treadmill exercise on glucose tolerance in diabetic rats.

Values are mean±standard error of mean for n=8 in each group. ZDF-Con, sedentary Zucker diabetic fatty; ZDF-Ex, exercised Zucker diabetic fatty; ZLC-Con, sedentary Zucker diabetic fatty lean control.

aP<0.05 compared with ZLC-Con group, bP<0.05 compared with ZDF-Con group.



(A) Blood glucose concentration was measured at various time points after dextrose (2 g/kg) treatment.

(B) Area under the curve (AUC) of intraperitoneal glucose tolerance test was calculated for quantified comparison.

Fig. Effect of 12 weeks of treadmill exercise on expression of interleukin-15 (IL-15) in soleus (SOL) and gastrocnemius medial (GM) muscles.

Values are mean±standard error of mean for n=8 in each group. ZLC-Con, sedentary Zucker diabetic fatty lean control; ZDF-Con, sedentary Zucker diabetic fatty; ZDF-Ex, exercised Zucker diabetic fatty. aP<0.05 compared with ZLC-Con group, bP<0.05 compared with ZDF-Con group.



Fig. Association of interleukin-15 (IL-15) expression in soleus (SOL) with body weight and fasting blood glucose levels.

Pearson correlation coefficients of IL-15 expression in SOL with (A) body weight and (B) concentration of fasting blood glucose are shown. Pearson correlation coefficients and P values are shown in each graph.



Introduction of myokine

Irisin Fat browning

FNDC5: fibronectin type III domain-containing protein 5

SP: signal peptide C: C-terminal domain Human mouse

10 weeks of endurance training

Irisin : brown fat like development of white fat

Kelly, Science, 2012

Mitochondria biogenesis UCP1 activation

Metabolic disease (Obesity)

Comparing the two-type of exercise (endurance vs. resistance) on circulating irisin level in obese adults

Aging (Sarcopenia)

Evaluating the effect of resistance exercise on skeletal muscle and circulating irisin level in aging mice and human.

Experimental results in SNU (irisin)

Metabolic disease model (human obesity)

Physiological Research, Accepted, July 2015

- To examined the effect of 12 weeks aerobic exercise training and resistance exercise training on circulating irisin level and body composition of obese adults

Purpose

Effect of aerobic training and resistance training on circulating irisin level and their association with change of body composition in obese

adults : a pilot study

Hee-jae Kim, Hyo-Joo Lee, Donghyun Yooon, Byunghun So, Junseok Son, Wook Song

Randomized

Control (Con, n= 7)

Post-Test Blood Collection After 8 h of Last intervention


Aerobic Exercise (AEX, n= 10) Resistance Exercise (REX, n= 10)

12 Weeks of Resistance Exercise (n= 10)

12 Weeks of Aerobic Exercise (n= 10)

Obese students (n= 27)

Pre-Test, Blood Collection

Outcome measurement : Leg Extensor, Flexor Strength (Isokinetic, Isometric) grip-strength, body composition (BIA) Irisin (ELISA)


The inclusion criteria required that a BMI >25kg/㎡, - Women : body fat percentage > 30% - Men : body fat percentage >25% .

Also, they required to have stable body weight (±2kg) for 3 month before enrollment, and a sedentary lifestyle (absence of moderate to vigorous regular physical activity in the past 3 month).

Irisin expression after 12 weeks of aerobic and resistance training in plasma


- Only 12 weeks of resistance exercise training increased blood irisin level significantly compared to aerobic training.

Physiological Research, Accepted, July 2015

Hee-jae Kim, Byunghun So, Mijung Choi, Dongheon Kang, Wook Song

Resistance exercise training increases the expression of irisin concomitant with improvement

of muscle function in aging mice and human

Aging (Sarcopenia) – animal & human study

Experimental Gerontology (Accepted, Jul 07, 2015)

Fourteen 19-month aged male mice (C57BL/6)

Old Resistance exercise group (ORT, n=8)

Old Control group (OCON, n=7)

12 weeks of Chronic Resistance Exercise (3days per week)

Body composition (DEX), Grip Strength Meter, Muscular endurance capacity, Mobility


Tissue Collection After 48 h of Last intervention

Aging (Sarcopenia) – animal study

Exercise Period Adaptation

(1wk) 1

Wk 2

Wk 3

Wk 4

Wk 5

Wk 6

Wk 7

Wk 8

Wk 9

Wk 10 Wk

11 Wk

12 Wk

Reps. 4 Rep Max.8

Load 0g 10% Body weight/ +2 gram for every 4th successful trails + 2 gram for every exercise session

Rest 2 min

Frequency 3 days/week

Animal

Randomized

Control group (n= 8) Not complete the study (n=6)

Post-Test Blood Collection After 8 h of Last intervention


Exercise group (n= 22) Not complete the study (n=2)

12 Weeks of Resistance Exercise (n= 22)

A total of 40 women with age over 65 years

Pre-Test, Blood Collection

Aging (Sarcopenia) – human study

Outcome measurement : Leg Extensor, Flexor Strength (Isokinetic, Isometric) grip-strength, body composition (BIA), blood maker Irisin (ELISA)

Human

Contents Intensity Warm up (10 mins) Dynamic/Static Stretching 15~60 sec.

Main exercise (40 mins)

Squat, Lunge, Bridge *HSPT group “Green” elastic band

RPE = 12 ~ 13 12-15 rep/ 3 set

*Control group

Stretching (static, dynamic) 1 hour / week

Leg Raise, Reverse Crunch, Sit-up, Crunch, Back Extension

Superman Position, Deadlift

Push-up, Back Row, Shoulder Press Biceps Curl, Triceps Extension

Cool down (10 mins) Dynamic/Static Stretching 15~60 sec.

High-speed power training (HSPT) protocol

Concentric contraction (“as fast as possible”)

paused ( 1 sec )

Eccentric contraction ( 2 sec )

Slow speed strength training (SSST) protocol

Concentric contraction ( 2-3 sec )

paused ( 1 sec )

Eccentric contraction ( 2-3 sec )

Exercise group : 60 min work out, 2 days/wk, 12 weeks

Control group : 60 min stretching, 1 day / wk, 12 weeks

(Sayers et al., 2010; Kang et al., 2014)

(Sayers et al., 2010)


Human


Animal

Basic characteristic of aging mice

Values are presented as mean ± SEM


There was no significant change in body composition or blood lipid profiles

Changes in characteristics of participants (human study)

Human

Aging (Sarcopenia) – animal & human study

Human Change of leg strength and grip strength after 12 weeks of resistance exercise in aging human



Muscle quality = grip strength / lean mass (DEXA)

Grip strength and Muscle quality

Effect of resistance exercise on improvement of physical functions in aging mice


Muscle endurance (Hanging test)



Mouse Physical Activity (Mobility) Test

Testing protocol - Duration – 30 min recording by video camera fixed above the device (dimmed room) - 1-hour adaption in the device prior to activity test Device - Acrylic made 100 cm x 100 cm x 16 cm - 21 holes in the cover plate were made for the ventilation - Each lined-square is 10 cm x 10 cm Counting Method - Activity score of 1 was counted when the body of a mouse crossed the line drawn above the device. - A diagonal crossing movement was counted as 2 in activity score.

Top view

Physical Activity Testing Device

100cm 100cm

16cm




Effect of resistance exercise on irisin expressions in serum and skeletal muscle in aging mice

The expressions of irisin were measured in (A) serum and (B) skeletal muscles including SOL, EDL, TA, and GAS after 12 weeks of progressive resistance exercise. Irisin expression in skeletal muscle was calculated by dividing total protein of each muscle tissues *p<0.05 compared with OLD-Con group. Values are mean ± S.E.M. for Old-Con (n=6) and Old-Ex (n=7).

Animal



Effect of resistance exercise on serum irisin expression and its correlation with improvement of muscle strength in aging human

The level of irisin in serum was measured after 12 weeks of resistance exercise training (A). The change of irisin expression in serum has positive correlations with improvement of grip strength (B) and isokinetic leg strength (C).

Values are mean ± S.E.M. for Old-Con (n=14) and Old-Ex (n=18).

Human


Summary

• Positive improvement of body composition has been resulted from 8 weeks progressive resistance training.

• Also, alternation of circulating irisin was highly correlated with increase of skeletal muscle and decrease of fat mass.

• 12 weeks progressive resistance training could increase the circulating irisin level in both aging mice and human subjects concomitant with improvement in muscle strength and muscle function.

• In addition, there were positive correlations between increases of circulating irisin level and improvement in muscle strength (both upper and lower body).

Limitation & Future study

• Further research is clearly required to identify how exercise-induced myokines are related to chronic or metabolic diseases including type 2 diabetes, osteoporosis, heart disease, and muscle atrophy.

• Based on these results, we provided positive correlations between circulation of myokines level and increase in skeletal muscle mass and strength.

• Especially, we need to identify the mechanism of crosstalk in organ to organ, for examples, adipose tissue to muscle, muscle to bone, and brain to skeletal muscle, using such ways like inhibitor, si-RNA and transgenic model.

Role of Myokines During Exercise

Potential mechanism for prevention of sarcopenia or aging related diseases

Integr Med Res, Review (BH So, HJ Kim, W Song et al., 2014)

IL-15: Interleukin-15 LIF: Leukemia inhibitory factor BDNF: Brain-derived neurotrophic factor FGF-21: Fibroblast growth factor 21 SPARC: Secreted protein acidic and rich in cysteine

Fat browning Muscle-fat crosstalk

Muscle hypertrophy

Metabolic diseases

Acknowledgment Funded by the Korean Government, Ministry of Education, Science, and Technology via Korea Research Foundation

Our Crew: Kim Heejae Yoon Donghyun Kim Janghoe So Byunghun Son Junseok Kim Jinsoo Song Hansol Kim Daeyoung Cho Suhan Hwang Suseung Lee Dongwon Kang Minji Choi Mijung Kang Donghun Kim Jiyeon Lee Hyojoo Woo Shinae Oh Seunglyul (post-doc) Kim Yongan (post-doc)

Collaborators: - Metabotyping of animal models for metabolic syndrome using

transcrptome and metabolomics : Je Kyung Seong, Dong-Mi Shin - Translational research for development of exercise mimetics for

control of sarcopenia : Kwon Ki Sun (KRIB), Sun G. Chung, Jae-Young Lim - The Korean sarcopenic obesity study : Kyung Mook Choi (Korea University)

Documents

Effect of Resistance Exercise on Myokine Expression in ... Song_Effect... · Kelly, Science, 2012 . Mitochondria biogenesis UCP1 activation . Metabolic disease (Obesity) Comparing