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Author Spotlight: Innovative Techniques for ROS Detection and Implications for Platelet Research

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Endothelial actin depolymerization mediates NADPH oxidase-superoxide production during flow reversal.

Jenny S Choy¹, Xiao Lu, Junrong Yang

¹Department of Biomedical Engineering, Indiana University Purdue University Indianapolis, Indianapolis, Indiana;

American Journal of Physiology. Heart and Circulatory Physiology

|November 5, 2013

Summary

Blood flow reversal increases superoxide production in vascular endothelial cells by depolymerizing actin filaments. This process involves NADPH oxidase, specifically the p47(phox) subunit, highlighting actin

Keywords:

endothelial cells

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Area of Science:

Cardiovascular Biology
Cellular Physiology
Biochemistry

Background:

Slow or reversed blood flow, characterized by negative wall shear stress, is linked to increased superoxide (O2(·-)) production in vascular endothelial cells.
The precise mechanisms by which shear stress influences O2(·-) production remain incompletely understood.

Purpose of the Study:

To investigate the hypothesis that actin depolymerization during flow reversal mediates O2(·-) production in vascular endothelial cells via NADPH oxidase, particularly the p47(phox) subunit.

Main Methods:

A swine model was used with complete blood flow reversal in one carotid artery as an experimental condition and forward flow in the contralateral artery as control.
Measurements included actin depolymerization, NADPH oxidase activity, reactive oxygen species (ROS) production, and protein levels/phosphorylation of NADPH oxidase subunits.
Pharmacological interventions such as jasplakinolide (actin stabilizer) and apocynin (antioxidant) were employed in vivo, and cytochalasin D was used in vitro to assess their effects.

Main Results:

Flow reversal induced actin depolymerization and a 3.9-fold increase in ROS production compared to forward flow.
NADPH oxidase activity increased by 1.4-fold in reversed flow segments, with increased p47(phox) phosphorylation but unchanged protein content.
In vivo treatment with jasplakinolide reduced the ROS increase to 1.7-fold, while apocynin completely prevented the ROS production. Cytochalasin D mimicked actin depolymerization and caused a 5.2-fold ROS increase.

Conclusions:

Actin filaments play a crucial role in mediating negative shear stress-induced ROS production in vascular endothelium.
The potentiation of NADPH oxidase activity, specifically involving the p47(phox) subunit, is a key mechanism in this process.
Targeting actin dynamics may offer therapeutic strategies for conditions associated with altered blood flow and oxidative stress.