The effect of reactive oxygen species on whole blood aggregation and the endothelial cell-platelet interaction in patients with coronary heart disease

Jonathan Watt, Marie-Ann Ewart, Fiona H Greig, Keith G Oldroyd, Roger M Wadsworth, Simon Kennedy

Research output: Contribution to journalArticle

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Abstract

Background
The effect of reactive oxygen species (ROS) on platelet function in coronary heart disease (CHD) is complex and poorly defined. Platelet aggregation studies in healthy volunteers have demonstrated contrasting results when platelets are exposed to ROS. We investigated the effect of ROS on whole blood aggregation (WBA) and the endothelial cell-platelet interaction in patients with CHD.

Methods and Results
ROS generated by xanthine and xanthine oxidase caused a concentration-dependent inhibition of WBA in blood from healthy donors and patients with CHD. In patients with CHD, 100 μM xanthine and 100 mU/ml xanthine oxidase inhibited WBA in response to 3 μg/ml collagen by 28.9% (95% CI 15.9%-41.8%, p < 0.001) and in response to 5 μM ADP by 36.0% (95% CI 9.6%-62.4%, p = 0.005). Using nitrotyrosine expression, platelets isolated from patients with CHD were found to be susceptible to peroxynitrite damage. The addition of 1 × 105 cultured endothelial cells inhibited WBA in response to 3 μg/ml collagen by 31.2% (95% CI 12.2%-50.2%, p < 0.05) and in response to 5 μM ADP by 31.6% (95% CI 2.5-60.7%, p < 0.05). Addition of xanthine and xanthine oxidase did not alter this effect, however pre-treatment of endothelial cells with a nitric oxide synthase inhibitor (L-NAME) partly reversed the inhibition.

Conclusion
ROS inhibit WBA in blood from patients with CHD. Whilst endothelial cells also inhibit WBA, the effect is attenuated by L-NAME, suggesting that nitric oxide is likely to remain an important protective mechanism against thrombosis in CHD.
Original languageEnglish
Pages (from-to)210-215
Number of pages6
JournalThrombosis Research
Volume130
Issue number2
DOIs
Publication statusPublished - Aug 2012

Fingerprint

Cell Communication
Coronary Disease
Reactive Oxygen Species
Blood Platelets
Endothelial Cells
Xanthine
Xanthine Oxidase
NG-Nitroarginine Methyl Ester
Adenosine Diphosphate
Collagen
Peroxynitrous Acid
Blood Donors
Platelet Aggregation
Nitric Oxide Synthase
Cultured Cells
Healthy Volunteers
Nitric Oxide
Thrombosis

Keywords

  • aged
  • blood platelets
  • cell communication
  • coronary disease
  • endothelial cells
  • endothelium, vascular
  • enzyme inhibitors
  • female
  • hemostasis
  • humans
  • male
  • middle aged
  • NG-nitroarginine methyl ester
  • platelet aggregation
  • reactive oxygen species
  • tyrosine
  • xanthine
  • xanthine oxidase

Cite this

The effect of reactive oxygen species on whole blood aggregation and the endothelial cell-platelet interaction in patients with coronary heart disease. / Watt, Jonathan; Ewart, Marie-Ann; Greig, Fiona H; Oldroyd, Keith G; Wadsworth, Roger M; Kennedy, Simon.

In: Thrombosis Research, Vol. 130, No. 2, 08.2012, p. 210-215.

Research output: Contribution to journalArticle

Watt, Jonathan ; Ewart, Marie-Ann ; Greig, Fiona H ; Oldroyd, Keith G ; Wadsworth, Roger M ; Kennedy, Simon. / The effect of reactive oxygen species on whole blood aggregation and the endothelial cell-platelet interaction in patients with coronary heart disease. In: Thrombosis Research. 2012 ; Vol. 130, No. 2. pp. 210-215.
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abstract = "BackgroundThe effect of reactive oxygen species (ROS) on platelet function in coronary heart disease (CHD) is complex and poorly defined. Platelet aggregation studies in healthy volunteers have demonstrated contrasting results when platelets are exposed to ROS. We investigated the effect of ROS on whole blood aggregation (WBA) and the endothelial cell-platelet interaction in patients with CHD.Methods and ResultsROS generated by xanthine and xanthine oxidase caused a concentration-dependent inhibition of WBA in blood from healthy donors and patients with CHD. In patients with CHD, 100 μM xanthine and 100 mU/ml xanthine oxidase inhibited WBA in response to 3 μg/ml collagen by 28.9{\%} (95{\%} CI 15.9{\%}-41.8{\%}, p < 0.001) and in response to 5 μM ADP by 36.0{\%} (95{\%} CI 9.6{\%}-62.4{\%}, p = 0.005). Using nitrotyrosine expression, platelets isolated from patients with CHD were found to be susceptible to peroxynitrite damage. The addition of 1 × 105 cultured endothelial cells inhibited WBA in response to 3 μg/ml collagen by 31.2{\%} (95{\%} CI 12.2{\%}-50.2{\%}, p < 0.05) and in response to 5 μM ADP by 31.6{\%} (95{\%} CI 2.5-60.7{\%}, p < 0.05). Addition of xanthine and xanthine oxidase did not alter this effect, however pre-treatment of endothelial cells with a nitric oxide synthase inhibitor (L-NAME) partly reversed the inhibition.ConclusionROS inhibit WBA in blood from patients with CHD. Whilst endothelial cells also inhibit WBA, the effect is attenuated by L-NAME, suggesting that nitric oxide is likely to remain an important protective mechanism against thrombosis in CHD.",
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author = "Jonathan Watt and Marie-Ann Ewart and Greig, {Fiona H} and Oldroyd, {Keith G} and Wadsworth, {Roger M} and Simon Kennedy",
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T1 - The effect of reactive oxygen species on whole blood aggregation and the endothelial cell-platelet interaction in patients with coronary heart disease

AU - Watt, Jonathan

AU - Ewart, Marie-Ann

AU - Greig, Fiona H

AU - Oldroyd, Keith G

AU - Wadsworth, Roger M

AU - Kennedy, Simon

N1 - Copyright © 2012 Elsevier Ltd. All rights reserved.

PY - 2012/8

Y1 - 2012/8

N2 - BackgroundThe effect of reactive oxygen species (ROS) on platelet function in coronary heart disease (CHD) is complex and poorly defined. Platelet aggregation studies in healthy volunteers have demonstrated contrasting results when platelets are exposed to ROS. We investigated the effect of ROS on whole blood aggregation (WBA) and the endothelial cell-platelet interaction in patients with CHD.Methods and ResultsROS generated by xanthine and xanthine oxidase caused a concentration-dependent inhibition of WBA in blood from healthy donors and patients with CHD. In patients with CHD, 100 μM xanthine and 100 mU/ml xanthine oxidase inhibited WBA in response to 3 μg/ml collagen by 28.9% (95% CI 15.9%-41.8%, p < 0.001) and in response to 5 μM ADP by 36.0% (95% CI 9.6%-62.4%, p = 0.005). Using nitrotyrosine expression, platelets isolated from patients with CHD were found to be susceptible to peroxynitrite damage. The addition of 1 × 105 cultured endothelial cells inhibited WBA in response to 3 μg/ml collagen by 31.2% (95% CI 12.2%-50.2%, p < 0.05) and in response to 5 μM ADP by 31.6% (95% CI 2.5-60.7%, p < 0.05). Addition of xanthine and xanthine oxidase did not alter this effect, however pre-treatment of endothelial cells with a nitric oxide synthase inhibitor (L-NAME) partly reversed the inhibition.ConclusionROS inhibit WBA in blood from patients with CHD. Whilst endothelial cells also inhibit WBA, the effect is attenuated by L-NAME, suggesting that nitric oxide is likely to remain an important protective mechanism against thrombosis in CHD.

AB - BackgroundThe effect of reactive oxygen species (ROS) on platelet function in coronary heart disease (CHD) is complex and poorly defined. Platelet aggregation studies in healthy volunteers have demonstrated contrasting results when platelets are exposed to ROS. We investigated the effect of ROS on whole blood aggregation (WBA) and the endothelial cell-platelet interaction in patients with CHD.Methods and ResultsROS generated by xanthine and xanthine oxidase caused a concentration-dependent inhibition of WBA in blood from healthy donors and patients with CHD. In patients with CHD, 100 μM xanthine and 100 mU/ml xanthine oxidase inhibited WBA in response to 3 μg/ml collagen by 28.9% (95% CI 15.9%-41.8%, p < 0.001) and in response to 5 μM ADP by 36.0% (95% CI 9.6%-62.4%, p = 0.005). Using nitrotyrosine expression, platelets isolated from patients with CHD were found to be susceptible to peroxynitrite damage. The addition of 1 × 105 cultured endothelial cells inhibited WBA in response to 3 μg/ml collagen by 31.2% (95% CI 12.2%-50.2%, p < 0.05) and in response to 5 μM ADP by 31.6% (95% CI 2.5-60.7%, p < 0.05). Addition of xanthine and xanthine oxidase did not alter this effect, however pre-treatment of endothelial cells with a nitric oxide synthase inhibitor (L-NAME) partly reversed the inhibition.ConclusionROS inhibit WBA in blood from patients with CHD. Whilst endothelial cells also inhibit WBA, the effect is attenuated by L-NAME, suggesting that nitric oxide is likely to remain an important protective mechanism against thrombosis in CHD.

KW - aged

KW - blood platelets

KW - cell communication

KW - coronary disease

KW - endothelial cells

KW - endothelium, vascular

KW - enzyme inhibitors

KW - female

KW - hemostasis

KW - humans

KW - male

KW - middle aged

KW - NG-nitroarginine methyl ester

KW - platelet aggregation

KW - reactive oxygen species

KW - tyrosine

KW - xanthine

KW - xanthine oxidase

U2 - 10.1016/j.thromres.2012.03.024

DO - 10.1016/j.thromres.2012.03.024

M3 - Article

VL - 130

SP - 210

EP - 215

JO - Thrombosis Research

JF - Thrombosis Research

SN - 0049-3848

IS - 2

ER -