Revealing the role of phosphatidylserine in shear stress-mediated protection in endothelial cells.
Previous studies have demonstrated that endothelial cells exposed to laminar shear stress are protected from apoptotic stimuli such as tumor necrosis factor (TNF)-alpha. The authors investigated the role of phosphatidylserine (PS) in this phenomenon. Western blot analysis of cleaved caspase 3 was used as an indicator of apoptosis and revealed that in the absence of serine, endothelial cells exposed to laminar shear stress were unable to protect against TNF-alpha-induced apoptosis, in contrast to sheared cells grown in regular medium. It was also found that shear-induced activation of the Akt pathway was significantly decreased in cells grown without serine. In addition, quantitation of PS using a novel isotopic labeling technique involving the use of formalin revealed that stearoyl-oleic PS (18:0/18:1) did not increase during shear treatment. These findings suggest that basal levels of PS are required to activate survival pathways in endothelial cells and thereby contribute to the overall protective mechanism initiated by shear stress.
Freed
Julie K
JK
Biotechnology and Bioengineering Center, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA.
Shortreed
Michael R
MR
Kleefisch
Christopher J
CJ
Smith
Lloyd M
LM
Greene
Andrew S
AS
D000818:Animals; D001783:Blood Flow Velocity; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D008055:Lipids / Q000302:isolation & purification / Q000502:physiology; D013058:Mass Spectrometry; D010718:Phosphatidylserines / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013314:Stress, Mechanical; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D008055:Lipids; D010718:Phosphatidylserines; D014409:Tumor Necrosis Factor-alpha