Cyclic strain induces an oxidative stress in endothelial cells.
Hypertension imposes an oxidant stress on the aorta and also causes mechanical deformation of the aortic wall. To assess whether deformation causes an oxidative stress, isolated porcine aortic endothelial cells (PAEC) were subjected to cyclic strain, and the cumulative amount of thiobarbituric acid reactive substances (TBARS, an index of lipid peroxidation) and H2O2 (a reactive oxygen species) was measured in the eluent at 2, 6, and 24 h. TBARS were increased by 40.5 +/- 9.2% after 24 h in cells exposed to cyclic strain vs. static controls (P < 0.05). No difference was seen at 2 and 6 h. H2O2 release was increased after 6 and 24 h of cyclic strain by 22.0 +/- 8.0 and 57.6 +/- 11.1 nmol H2O2/mg, respectively (P < 0.005), but was not increased after 2 h of strain. In vascular smooth muscle cells, TBARS were not observed and H2O2 release was not increased by cyclic strain. To investigate a potential source of H2O2 induced by strain, the activity of NADH/NADPH oxidase, a superoxide-generating enzyme, was measured by chemiluminescence. After 2 h, cells exposed to cyclic strain had greater activity than static controls (531.0 +/- 68.4 vs. 448.3 +/- 54.2 pmol O2- x mg(-1) x s(-1), respectively, when incubated with NADH, P < 0.005; 85.8 +/- 8.9 vs. 71.6 +/- 3.8 pmol O2- x mg(-1) x s(-1) when incubated with NADPH, P < 0.05). No effect on NADH/NADPH oxidase activity was seen after 6 or 24 h. The following conclusions were made: 1) cyclic strain induces an oxidant stress in PAEC monolayers as measured by TBARS formation and H2O2 release, 2) NADH/NADPH oxidase is a potential source of H2O2 release in cyclically strained cells, and 3) mechanical deformation of endothelial cells may play a critical role in the generation of oxidative stress within the vessel wall.
Howard
A B
AB
Division of Cardiology, Emory University, Atlanta, Georgia 30322, USA.
Alexander
R W
RW
Nerem
R M
RM
Griendling
K K
KK
Taylor
W R
WR
D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D004798:Enzymes / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D018384:Oxidative Stress; D010507:Periodicity; D013314:Stress, Mechanical; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013552:Swine; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism
D004798:Enzymes; D013438:Sulfhydryl Compounds; D017392:Thiobarbituric Acid Reactive Substances; D006861:Hydrogen Peroxide; D009247:NADH, NADPH Oxidoreductases