Aerobic Exercise, Other Healthy Lifestyle Factors and Vascular Aging Doug Seals Department of Integrative Physiology University of Colorado Boulder

I. Background and Significance II. Large Elastic Artery Stiffness, Aging and Exercise Functional Effects Mechanisms III. Endothelial Function, Aging and Exercise Functional Effects Mechanisms IV. Exercise: Vascular Protection from “Adverse Factors” V. Other Healthy Lifestyle Factors and Vascular Aging

3 The growing problem of cardiovascular diseases, aging and arterial dysfunction… Photo of the aging of man. From insightbrookline.com/blog/hey-old- manby-elliott-kronenfeld insightbrookline.com/blog/hey-old- manby-elliott-kronenfeld

Parkinson’s Disease AccidentsKidney Disease DiabetesAlzheimer’s Disease Respiratory Diseases Cancer CVD AHA 2012 From data published by CDC at cdc.gov/nchs/fastats/leading- causes-of-death.htm cdc.gov/nchs/fastats/leading- causes-of-death.htm

Prevalence of CVD of All Causes in Relation to Age Over 90% of CVD occurs in middle-aged and older adults

… should we be bracing for a new epidemic of ‘boomer’-driven CVD?

By 2030: 40% of U.S. adults will have >1 form of CVD Medical costs of CVD will triple These changes will be driven largely by the aging of the population “… Effective prevention strategies are needed if we are to limit the growing burden of (age-associated) CVD.” Circulation 123:933, 2011 Article freely available at circ.ahajournals.org/content/123/8/933.full circ.ahajournals.org/content/123/8/933.full

Aging CVD How does aging CVD risk?

Aging CVD Dysfunction of Arteries

10 2 key changes with arterial aging

endothelial dysfunction 2 key changes with arterial aging NO nitric oxide

CVD ↓endothelial function ↑arterial stiffness }{ Dysfunction of Arteries Aging cognitive dysfunction, brain diseases of aging, etc.

CVD ↓endothelial function ↑arterial stiffness }{ Dysfunction of Arteries Lifestyle, Pharmacological, Nutraceutical Strategies Aging

Regular Aerobic Exercise Reviews: Seals et al.: J Appl Physiol 105: , 2008J Appl Physiol 105: , 2008 J Physiol 587: , 2009J Physiol 587: , 2009 Clin Sci 120:357-75, 2011Clin Sci 120:357-75, 2011 Physiology 29:250-64, 2014Physiology 29:250-64, 2014 DeVan & Seals Exp Physiol 97:305-10, 2012 Experimental work:

I. Background and Significance II. Large Elastic Artery Stiffness, Aging and Exercise Functional Effects Mechanisms III. Endothelial Function, Aging and Exercise Functional Effects Mechanisms IV. Exercise: Vascular Protection from “Adverse Factors” V. Other Healthy Lifestyle Factors and Vascular Aging

CVD ↓endothelial function ↑arterial stiffness }{ Dysfunction of Arteries Aging Aerobic Exercise ??

aorta Aortic Stiffness: Aortic Pulse Wave Velocity (aPWV) increasing age

Assessment of aPWV velocity = time distance time

Attenuated Aortic Stiffening in Exercising Postmenopausal Women Figs. 1 & 2, upper right-hand figures. From Tanaka et al. Arterioscler Thromb Vasc Biol 18:127-32, Freely available at atvb.ahajournals.org/content/18/1/127.full atvb.ahajournals.org/content/18/1/127.full

Ultrasound Probe Tonometer diameter carotid bulb ∆ pressure compliance = ∆ pressure ∆ diameter Assessment of Carotid Artery Compliance

* * ** Better Preserved Carotid Compliance in Exercising Adults Data from Tanaka et al. Circ 102:1270-5, 2000 andTanaka et al. Circ 102:1270-5, 2000 Moreau et al. Cardiovasc Res 57:861-68, 2003Cardiovasc Res 57:861-68, 2003

Several Types of Aerobic Exercise Preserve Carotid Compliance Data from Tanaka and colleagues and DeVan and Seals. Exp Physiol 97:305-10, 2012Exp Physiol 97:305-10, 2012

Aerobic Exercise Intervention Improves Carotid Compliance Walking min/d ~6 d/wk 8-12 wks 70-75% of maximal HR Compliance: HR monitors Fig. 2a. From Moreau et al. Cardiovasc Res 57:861-68, Freely available at cardiovascres.oxfordjournals.org/ content/57/3/861.full See also Tanaka et al., Circ 102:1270-5, 2000Circ 102:1270-5, 2000.

Mechanisms Aerobic Exercise and Arterial Stiffness

25 The need for “pre-clinical” models...

aPWV = d/t pulse wave Aortic Stiffness in Vivo: Aortic Pulse Wave Velocity (aPWV) in Mice * Data from Fleenor and colleagues. Aging Cell 10: , 2011 ( Aging Cell 11: , 2012 ( Exp Gerontol 47:588-94, 2012 (

Old Voluntary Running Pre Old Voluntary Running Post * In Vivo Validation of Model: Voluntary Wheel Running Reduces aPWV in Old Mice Gioscia-Ryan et al. Unpublished data.

oxidative stress Modulation of Oxidative Stress aging... nitrotyrosine

Wheel Running Reverses Aortic Oxidative Stress Fig 3. From Durrant et al. J Physiol 587: , 2009 Freely available at jp.physoc.org/content/587/13/3271.full jp.physoc.org/content/587/13/3271.full

Arterial Wall Structural Modifications elastin collagen AGEs aging... ?

Collagen I Expression in Carotid Arteries (IHC) Fig. 2B. From Fleenor et al. J Physiol 588: , Freely available at jp.physoc.org/content/588/20/3971.full jp.physoc.org/content/588/20/3971.full

AGEs Advanced Glycation Endproducts (AGEs) in Aorta Young Control Old Control Old Voluntary Running * Jason Eng, unpublished data

Summary/Conclusions

I. Background and Significance II. Large Elastic Artery Stiffness, Aging and Exercise Functional Effects Mechanisms III. Endothelial Function, Aging and Exercise Functional Effects Mechanisms IV. Exercise: Vascular Protection from “Adverse Factors” V. Other Healthy Lifestyle Factors and Vascular Aging

CVD ↓endothelial function ↑arterial stiffness }{ Dysfunction of Arteries Aging Aerobic Exercise ??

Dilation produced by a stimulus that evokes NO synthesis/release from vascular endothelium Endothelium-dependent dilation (EDD) Assessment of Endothelial Function stimulus youngerolder NO nitric oxide

Endothelium-Dependent Dilation

brachial artery catheter pharmacological infusions forearm strain gauge ECG, BP Chemical Stimulation of EDD Saline ACh

Aerobic Exercise Preserves Endothelial Function with Aging in Men (ACh) ? ? Fig. 1, upper left. From DeSouza et al. Circ 102: , Freely available at circ.ahajournals. org/content/102 /12/1351.full circ.ahajournals. org/content/102 /12/1351.full Fig. 1, upper right. From DeSouza et al. Circ 102: , Freely available at circ.ahajournals.org/content/10 2/12/1351.full circ.ahajournals.org/content/10 2/12/1351.full Fig. 2, upper left. From DeSouza et al. Circ 102: , 2000 Freely available at circ.ahajournals.org/content/10 2/12/1351.full circ.ahajournals.org/content/10 2/12/1351.full Data from Taddei et al. Circ 101: , Freely available at circ.ahajournals.org/content/101/ 25/2896.full?sid=4359e86d d80-b4ab-69d4a4bb4cd0 circ.ahajournals.org/content/101/ 25/2896.full?sid=4359e86d d80-b4ab-69d4a4bb4cd0

Ultrasound Imaging record: inflate cuff deflate cuff vs. Blood Flow Stimulation of EDD (Brachial Artery Flow-Mediated Dilation) inflated

Aerobic Exercise Preserves Endothelial Function with Aging in Men (FMD) Data from Eskurza et al. J Physiol 556:315-24, Freely available at jp.physoc.org/content/556/1/315.fulljp.physoc.org/content/556/1/315.full * Young Non-Ex Older Non-Ex Older Exercise ? ? Fig. 1, middle right panel for men. From Pierce et al. Clin Sci 120:13-23, Freely available at clinsci.org/cs/ev/120/0 013/cs _ev.htm clinsci.org/cs/ev/120/0 013/cs _ev.htm

42 “Macro”-Mechanisms of Aging and Exercise on Endothelial Function

O2-O2- NADPH Oxidase SOD NO eNOS relaxation NF-kB ONOO- ? Oxidative Stress and Inflammation: Key Events inflammatory cytokines

Influence of Superoxide on EDD with Aging and Exercise in Men * Saline Older Non-Ex Older Exercise Young Non-Ex Vitamin C TEMPOL Young Non-Ex Older Exercise Older Non-Ex Data modified from Eskurza et al. J Physiol 556:315-24, Freely available at jp.physoc.org/content/556/1/315.full jp.physoc.org/content/556/1/315.full Data modified from Durrant et al. J Physiol 587: , 2009 Freely available at jp.physoc.org/content/587/13/3271.full jp.physoc.org/content/587/13/3271.full Data modified from Taddei et al. Circ 101: , Freely available at circ.ahajournals.org/content /101/25/2896.full?sid=4359 e86d d80-b4ab- 69d4a4bb4cd0 circ.ahajournals.org/content /101/25/2896.full?sid=4359 e86d d80-b4ab- 69d4a4bb4cd0

Oxidative Stress: Nitrotyrosine Abundance

46 Endothelial Cell Acquisition/Analysis in Human Subjects (Clinical Research Center) Work by Donato, LaRocca and Kaplon

Oxidative Stress in Mice and Humans: Direct Molecular Evidence Fig. 3 (left 3 bars and gels only). From Durrant et al. J Physiol 587: , 2009 Freely available at jp.physoc.org/content/587/ 13/3271.full jp.physoc.org/content/587/ 13/3271.full Fig. 1A. From Pierce et al. Aging Cell 10: , Freely available at onlinelibrary.wiley.com/doi/ /j x/full onlinelibrary.wiley.com/doi/ /j x/full

O2-O2- NADPH Oxidase SOD NO eNOS relaxation NF-kB ONOO- inflammatory cytokines Oxidative Stress: NADPH Oxidase * Young Control Old Control Old Voluntary Running ? Apocynin NADPH Oxidase Data from Durrant et al. J Physiol 587: , 2009 Freely available at jp.physoc.org/content/587/13/3271.fulljp.physoc.org/content/587/13/3271.full

NADPH Oxidase in Mouse and Human Arteries Fig. 5A (left 3 columns and gels only). From Durrant et al. J Physiol 587: , Freely available at jp.physoc.org/content/587/13/3271.full jp.physoc.org/content/587/13/3271.full Fig. 1B. From Pierce et al. Aging Cell 10: , Freely available at onlinelibrary.wiley.com/doi/ /j x/full onlinelibrary.wiley.com/doi/ /j x/full

O2-O2- NADPH Oxidase SOD NO eNOS relaxation NF-kB ONOO- inflammatory cytokines ? Modulation of Superoxide Dismutase

Superoxide Dismutase in Mouse and Human Arteries Fig. 4B (Y CC, O CC, and O VR only). From Durrant et al. J Physiol 587: , Freely available at jp.physoc.org/content/587/13/3271.full jp.physoc.org/content/587/13/3271.full Fig. 1D. From Pierce et al. Aging Cell 10: , Freely available at onlinelibrary.wiley.com/doi/ /j x/full onlinelibrary.wiley.com/doi/ /j x/full

Extracellular Superoxide Dismutase... heparin circulation... ecSO D Fig. 2. From Pierce et al. Aging Cell 10: , Freely available at onlinelibrary.wiley.com/doi/ /j x/full onlinelibrary.wiley.com/doi/ /j x/full

Role of Inflammation O2-O2- NADPH Oxidase SOD NO eNOS relaxation NF-kB ONOO- ?

IL-6 IFN- ɣ TNF-α * * * Young Control Old Control Old Vol. Running Young Control Old Control Old Vol. Running Young Control Old Control Old Vol. Running Anti-inflammatory Effects of Exercise with Arterial Aging in Mice Fig. 2, B-D. From Lesniewski et al. Am J Physiol 301:H1025-H1032, Reprinted with permission. Freely available at ajpheart.physiology.org/content/301/3/H1025 ajpheart.physiology.org/content/301/3/H1025

NF-kB Effects of Exercise on Endothelial Inflammation with Aging in Humans ? IL-6 Fig. 1C. From Pierce et al. Aging Cell 10: , Freely available at onlinelibrary.wiley.com/doi/ /j x/full onlinelibrary.wiley.com/doi/ /j x/full Kaplon, unpublished data. Fig. 4A. From Donato et al. Aging Cell 7:805-12, Freely available at onlinelibrary.wiley.com/ doi/ /j x/full onlinelibrary.wiley.com/ doi/ /j x/full

NF-kB Anti-inflammatory Effects of Exercise on Endothelial Function with Aging * Data from Walker et al. Clin Sci 127: , 2014 Freely available at clinsci.org/cs/127/0645/cs htmclinsci.org/cs/127/0645/cs htm

CVD ↓endothelial function ↑arterial stiffness }{ Dysfunction of Arteries Aging Aerobic Exercise Summary/Conclusions O2-O2- NADPH Oxidase NO NF-kB SOD

I. Background and Significance II. Large Elastic Artery Stiffness, Aging and Exercise Functional Effects Mechanisms III. Endothelial Function, Aging and Exercise Functional Effects Mechanisms IV. Exercise: Vascular Protection from “Adverse Factors” V. Other Healthy Lifestyle Factors and Vascular Aging

Protecting Against vs. Lowering Risk Factors for CVD Increasing age Male sex Heredity Smoking High blood cholesterol High blood pressure Physical inactivity Obesity and overweight Diabetes mellitus Stress Excessive alcohol consumption Aerobic Exercise ? ?

Exercise-Associated “Resistance” to LDL-Cholesterol Fig 1a. From Walker et al. Am J Hypertens 22:250-6, Available at ajh.oxfordjournals.org/conte nt/22/3/250.full ajh.oxfordjournals.org/conte nt/22/3/250.full Fig 1b. From Walker et al. Am J Hypertens 22:250-6, Available at ajh.oxfordjournals.org/conte nt/22/3/250.full ajh.oxfordjournals.org/conte nt/22/3/250.full

Increasing age Male sex (gender) Heredity (including Race) Smoking High blood cholesterol High blood pressure Physical inactivity Obesity and overweight Diabetes mellitus Stress Excessive alcohol consumption Aerobic Exercise ? ?

Exercise-Associated “Resistance” to Pre-Diabetes Normal FBG (NFG) = <100 mg/dl Impaired FBG (IFG) = mg/dl Data from DeVan et al. Clin Sci 124:325-31, Freely available at clinsci.org/cs/124/0325 /cs htm * **

I. Background and Significance II. Large Elastic Artery Stiffness, Aging and Exercise Functional Effects Mechanisms III. Endothelial Function, Aging and Exercise Functional Effects Mechanisms IV. Exercise: Vascular Protection from “Adverse Factors” V. Other Healthy Lifestyle Factors and Vascular Aging

Abdominal adiposity arterial function High Risk: >35 in. >40 in. weight loss arterial function

Dietary sodium intake teaspoon 2300 mg Average daily intake (US) 3400 mg Dietary guidelines: 1500 mg ↑70% ↓25% vs.

Other ways to promote arterial health with aging Lifestyle ✓ Nutritional approaches…? Pharmacological…? ?

Good evidence: diet pharmacological “nutraceutical”

Under study: nitrate/nitrite vitamin D MitoQ trehalose (autophagy) NO boosting curcumin turmeric

Summary and Conclusions

“Aging” of arteries is a (the) major risk factor for the development of CVD and will be an even greater biomedical threat/challenge in the future Conclusions

Regular aerobic exercise is an effective strategy for preserving arterial function and health with aging, and this is mediated (at least in part) by suppression of vascular oxidative stress and inflammation Conclusions

Regular aerobic exercise may exert these beneficial effects partly by protecting aging arteries from multiple “adverse factors” Conclusions

Other healthy lifestyle factors also have a major influence on arterial function with aging, and “healthy lifestyle-mimicking” pharmaceutical and nutraceutical compounds hold promise as complementary strategies

Extramural Funding R37 (MERIT) AG R01 AG T32 AG R01 HL107120