Elevated arterial shear rate increases indexes of endothelial cell autophagy and nitric oxide synthase activation in humans

Seul Ki Park, D. Taylor La Salle, James Cerbie, Jae Min Cho, Amber Bledsoe, Ashley Nelson, David E. Morgan, Russell S Richardson, Yan-Ting Shiu, Sihem Boudina, Joel D. Trinity, John David Symons

Research output: Contribution to journalArticle

Abstract

Continuous laminar shear stress increases the process of autophagy, activates endothelial nitric oxide (NO) synthase phosphorylation at serine 1177 (p-eNOSS1177), and generates NO in bovine and human arterial endothelial cells (ECs) compared with static controls. However, the translational relevance of these findings has not been explored. In the current study, primary ECs were collected from the radial artery of 7 men using sterile J-wires before (Pre) and after (Post) 60 min of rhythmic handgrip exercise (HG) performed with the same arm. After ECs were identified by positive costaining for vascular endothelial cadherin and 4',6'-diamidino-2-phenylindole, immunofluorescent antibodies were used to assess indices of autophagy, NO generation, and superoxide anion (O2·-) production. Commercially available primary human arterial ECs were stained and processed in parallel to serve as controls. All end points were evaluated using 75 ECs from each subject. Relative to Pre-HG, HG elevated arterial shear rate ( P < 0.05) ~3-fold, whereas heart rate, arterial pressure, and cardiac output were not altered. Compared with values obtained from ECs Pre-HG, Post-HG ECs displayed increased ( P < 0.05) expression of p-eNOSS1177, NO generation, O2·- production, BECLIN1, microtubule-associated proteins 1A/1B light chain 3B, autophagy-related gene 3, and lysosomal-associated membrane protein 2A and decreased ( P < 0.05) expression (i.e., enhanced degradation) of the adaptor protein p62/sequestosome-1. These novel findings provide evidence that elevated arterial shear rate associated with functional hyperemia initiates autophagy, activates p-eNOSS1177, and increases NO and O2·- generation in primary human ECs. NEW & NOTEWORTHY Previously, our group reported in bovine arterial and human arterial endothelial cells (ECs) that shear stress initiates trafficking of the autophagosome to the lysosome and increases endothelial nitric oxide (NO) synthase phosphorylation at serine 1177, NO generation, and O2·- production. Here, the translational relevance of these findings is documented. Specifically, functional hyperemia induced by rhythmic handgrip exercise elevates arterial shear rate to an extent that increases indices of autophagy, NO generation, and O2·- production in primary arterial ECs collected from healthy men.

LanguageEnglish (US)
PagesH106-H112
JournalAmerican journal of physiology. Heart and circulatory physiology
Volume316
Issue number1
DOIs
StatePublished - Jan 1 2019

Fingerprint

Autophagy
Nitric Oxide Synthase
Fluorescent Antibody Technique
Blood Vessels
Endothelial Cells
Nitric Oxide
Exercise
Nitric Oxide Synthase Type III
Hyperemia
Lysosomal-Associated Membrane Protein 3
Serine
Phosphorylation
Radial Artery
Lysosomes
Superoxides
Cardiac Output
Arterial Pressure
Heart Rate
Light
Antibodies

Keywords

  • blood vessel
  • exercise
  • immunofluorescence
  • shear stress

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

Cite this

Elevated arterial shear rate increases indexes of endothelial cell autophagy and nitric oxide synthase activation in humans. / Park, Seul Ki; La Salle, D. Taylor; Cerbie, James; Cho, Jae Min; Bledsoe, Amber; Nelson, Ashley; Morgan, David E.; Richardson, Russell S; Shiu, Yan-Ting; Boudina, Sihem; Trinity, Joel D.; Symons, John David.

In: American journal of physiology. Heart and circulatory physiology, Vol. 316, No. 1, 01.01.2019, p. H106-H112.

Research output: Contribution to journalArticle

Park, Seul Ki ; La Salle, D. Taylor ; Cerbie, James ; Cho, Jae Min ; Bledsoe, Amber ; Nelson, Ashley ; Morgan, David E. ; Richardson, Russell S ; Shiu, Yan-Ting ; Boudina, Sihem ; Trinity, Joel D. ; Symons, John David. / Elevated arterial shear rate increases indexes of endothelial cell autophagy and nitric oxide synthase activation in humans. In: American journal of physiology. Heart and circulatory physiology. 2019 ; Vol. 316, No. 1. pp. H106-H112.
@article{cd1f812e9e5544449063c5fc0b01a4d5,
title = "Elevated arterial shear rate increases indexes of endothelial cell autophagy and nitric oxide synthase activation in humans",
abstract = "Continuous laminar shear stress increases the process of autophagy, activates endothelial nitric oxide (NO) synthase phosphorylation at serine 1177 (p-eNOSS1177), and generates NO in bovine and human arterial endothelial cells (ECs) compared with static controls. However, the translational relevance of these findings has not been explored. In the current study, primary ECs were collected from the radial artery of 7 men using sterile J-wires before (Pre) and after (Post) 60 min of rhythmic handgrip exercise (HG) performed with the same arm. After ECs were identified by positive costaining for vascular endothelial cadherin and 4',6'-diamidino-2-phenylindole, immunofluorescent antibodies were used to assess indices of autophagy, NO generation, and superoxide anion (O2·-) production. Commercially available primary human arterial ECs were stained and processed in parallel to serve as controls. All end points were evaluated using 75 ECs from each subject. Relative to Pre-HG, HG elevated arterial shear rate ( P < 0.05) ~3-fold, whereas heart rate, arterial pressure, and cardiac output were not altered. Compared with values obtained from ECs Pre-HG, Post-HG ECs displayed increased ( P < 0.05) expression of p-eNOSS1177, NO generation, O2·- production, BECLIN1, microtubule-associated proteins 1A/1B light chain 3B, autophagy-related gene 3, and lysosomal-associated membrane protein 2A and decreased ( P < 0.05) expression (i.e., enhanced degradation) of the adaptor protein p62/sequestosome-1. These novel findings provide evidence that elevated arterial shear rate associated with functional hyperemia initiates autophagy, activates p-eNOSS1177, and increases NO and O2·- generation in primary human ECs. NEW & NOTEWORTHY Previously, our group reported in bovine arterial and human arterial endothelial cells (ECs) that shear stress initiates trafficking of the autophagosome to the lysosome and increases endothelial nitric oxide (NO) synthase phosphorylation at serine 1177, NO generation, and O2·- production. Here, the translational relevance of these findings is documented. Specifically, functional hyperemia induced by rhythmic handgrip exercise elevates arterial shear rate to an extent that increases indices of autophagy, NO generation, and O2·- production in primary arterial ECs collected from healthy men.",
keywords = "blood vessel, exercise, immunofluorescence, shear stress",
author = "Park, {Seul Ki} and {La Salle}, {D. Taylor} and James Cerbie and Cho, {Jae Min} and Amber Bledsoe and Ashley Nelson and Morgan, {David E.} and Richardson, {Russell S} and Yan-Ting Shiu and Sihem Boudina and Trinity, {Joel D.} and Symons, {John David}",
year = "2019",
month = "1",
day = "1",
doi = "10.1152/ajpheart.00561.2018",
language = "English (US)",
volume = "316",
pages = "H106--H112",
journal = "American Journal of Physiology - Heart and Circulatory Physiology",
issn = "0363-6135",
publisher = "American Physiological Society",
number = "1",

}

TY - JOUR

T1 - Elevated arterial shear rate increases indexes of endothelial cell autophagy and nitric oxide synthase activation in humans

AU - Park, Seul Ki

AU - La Salle, D. Taylor

AU - Cerbie, James

AU - Cho, Jae Min

AU - Bledsoe, Amber

AU - Nelson, Ashley

AU - Morgan, David E.

AU - Richardson, Russell S

AU - Shiu, Yan-Ting

AU - Boudina, Sihem

AU - Trinity, Joel D.

AU - Symons, John David

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Continuous laminar shear stress increases the process of autophagy, activates endothelial nitric oxide (NO) synthase phosphorylation at serine 1177 (p-eNOSS1177), and generates NO in bovine and human arterial endothelial cells (ECs) compared with static controls. However, the translational relevance of these findings has not been explored. In the current study, primary ECs were collected from the radial artery of 7 men using sterile J-wires before (Pre) and after (Post) 60 min of rhythmic handgrip exercise (HG) performed with the same arm. After ECs were identified by positive costaining for vascular endothelial cadherin and 4',6'-diamidino-2-phenylindole, immunofluorescent antibodies were used to assess indices of autophagy, NO generation, and superoxide anion (O2·-) production. Commercially available primary human arterial ECs were stained and processed in parallel to serve as controls. All end points were evaluated using 75 ECs from each subject. Relative to Pre-HG, HG elevated arterial shear rate ( P < 0.05) ~3-fold, whereas heart rate, arterial pressure, and cardiac output were not altered. Compared with values obtained from ECs Pre-HG, Post-HG ECs displayed increased ( P < 0.05) expression of p-eNOSS1177, NO generation, O2·- production, BECLIN1, microtubule-associated proteins 1A/1B light chain 3B, autophagy-related gene 3, and lysosomal-associated membrane protein 2A and decreased ( P < 0.05) expression (i.e., enhanced degradation) of the adaptor protein p62/sequestosome-1. These novel findings provide evidence that elevated arterial shear rate associated with functional hyperemia initiates autophagy, activates p-eNOSS1177, and increases NO and O2·- generation in primary human ECs. NEW & NOTEWORTHY Previously, our group reported in bovine arterial and human arterial endothelial cells (ECs) that shear stress initiates trafficking of the autophagosome to the lysosome and increases endothelial nitric oxide (NO) synthase phosphorylation at serine 1177, NO generation, and O2·- production. Here, the translational relevance of these findings is documented. Specifically, functional hyperemia induced by rhythmic handgrip exercise elevates arterial shear rate to an extent that increases indices of autophagy, NO generation, and O2·- production in primary arterial ECs collected from healthy men.

AB - Continuous laminar shear stress increases the process of autophagy, activates endothelial nitric oxide (NO) synthase phosphorylation at serine 1177 (p-eNOSS1177), and generates NO in bovine and human arterial endothelial cells (ECs) compared with static controls. However, the translational relevance of these findings has not been explored. In the current study, primary ECs were collected from the radial artery of 7 men using sterile J-wires before (Pre) and after (Post) 60 min of rhythmic handgrip exercise (HG) performed with the same arm. After ECs were identified by positive costaining for vascular endothelial cadherin and 4',6'-diamidino-2-phenylindole, immunofluorescent antibodies were used to assess indices of autophagy, NO generation, and superoxide anion (O2·-) production. Commercially available primary human arterial ECs were stained and processed in parallel to serve as controls. All end points were evaluated using 75 ECs from each subject. Relative to Pre-HG, HG elevated arterial shear rate ( P < 0.05) ~3-fold, whereas heart rate, arterial pressure, and cardiac output were not altered. Compared with values obtained from ECs Pre-HG, Post-HG ECs displayed increased ( P < 0.05) expression of p-eNOSS1177, NO generation, O2·- production, BECLIN1, microtubule-associated proteins 1A/1B light chain 3B, autophagy-related gene 3, and lysosomal-associated membrane protein 2A and decreased ( P < 0.05) expression (i.e., enhanced degradation) of the adaptor protein p62/sequestosome-1. These novel findings provide evidence that elevated arterial shear rate associated with functional hyperemia initiates autophagy, activates p-eNOSS1177, and increases NO and O2·- generation in primary human ECs. NEW & NOTEWORTHY Previously, our group reported in bovine arterial and human arterial endothelial cells (ECs) that shear stress initiates trafficking of the autophagosome to the lysosome and increases endothelial nitric oxide (NO) synthase phosphorylation at serine 1177, NO generation, and O2·- production. Here, the translational relevance of these findings is documented. Specifically, functional hyperemia induced by rhythmic handgrip exercise elevates arterial shear rate to an extent that increases indices of autophagy, NO generation, and O2·- production in primary arterial ECs collected from healthy men.

KW - blood vessel

KW - exercise

KW - immunofluorescence

KW - shear stress

UR - http://www.scopus.com/inward/record.url?scp=85058768651&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85058768651&partnerID=8YFLogxK

U2 - 10.1152/ajpheart.00561.2018

DO - 10.1152/ajpheart.00561.2018

M3 - Article

VL - 316

SP - H106-H112

JO - American Journal of Physiology - Heart and Circulatory Physiology

T2 - American Journal of Physiology - Heart and Circulatory Physiology

JF - American Journal of Physiology - Heart and Circulatory Physiology

SN - 0363-6135

IS - 1

ER -