CYCLIC STRAIN INDUCES AN OXIDATIVE STRESS IN ENDOTHELIAL-CELLS

Hypertension imposes an oxidant stress on the aorta and also causes me chanical 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 (TEARS, an index of lipid pero xidation) and H2O2 (a reactive oxygen species) was measured in the elu ent at 2, 6, and 24 h. TEARS 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, TEARS were not observed and H2O2 re lease was not increased by cyclic strain. To investigate a potential s ource of H2O2 induced by strain, the activity of NADH/NADPH oxidase, a superoxide-generating enzyme, was measured by chemiluminescence. Afte r 2 h, cells exposed to cyclic strain had greater activity than static controls (531.0 +/- 68.4 vs. 448.3 +/- 54.2 pmol O-2(-) . mg(-1) . s( -1), respectively, when incubated with NADH, P < 0.005; 85.5 +/- 8.9 v s. 71.6 +/- 3.8 pmol O-2(-) . mg(-1) . 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: I) cyclic strain induce s an oxidant stress in PAEC monolayers as measured by TEARS 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 oxida tive stress within the vessel wall.