MAPK signaling mediates low shear stress-induced oxidative damage in human umbilical vein endothelial cells in vitro.
OBJECTIVE
Atherosclerotic lesions occur preferentially in the arterial branches, bifurcations and curvatures where shear stress is low. We aimed to study the possible mechanisms involved in low shear stress (LSS)-induced oxidative damage in vascular endothelial cells.


METHODS
Human umbilical vein endothelial cells (HUVECs) exposed for 60 min to simulated LSS using a parallel-plate flow chamber were examined for intracellular reactive oxygen species (ROS) and cell apoptosis with chemiluminescence assay and TUNEL staining, respectively. Western blotting was used to determine the levels of endothelial nitric oxide synthase (eNOS), P38, extracellular signal-regulate kinase (ERK) and c-Jun as well as their phosphorylation in cells with LSS exposure for different time lengths. To investigate the signaling pathway involved in LSS-induced oxidative damage, the cells were treated with P38, ERK and c-Jun inhibitors and examined for the expression of eNOS-Thr495 that negatively regulated eNOS.


RESULTS
Exposure to LSS for 1 h resulted in markedly increased ROS accumulation and apoptosis in HUVECs. LSS exposure time-dependently enhanced the phosphorylation of eNOS, P38, ERK and c-Jun but did not significantly affect their total protein expressions. Inhibition of ERK with PD98059 deactivated eNOS-Thr495 and restored super oxide dismutase (SOD) activity, while inhibition of either p38 with SB202190 or c-Jun with SP600125 did no produce such effects.


CONCLUSIONS
LSS-induced oxidative damage is partly due to activated mitogen-activated protein kinases (MAPK), among which ERK contributes to decreased NO release in endothelial cells.
Department of Cardiology, Nanjing First Hospital, Nanjing Medical University, Nanjing 210006, China. E-mail: wzm434@126.com.
Zhimei
Z
Wang
Junxia
J
Zhang
Bing
B
Li
Wenxing
W
Mao
Shaoliang
S
Chen
D017209:Apoptosis; D002478:Cells, Cultured; D005419:Flavonoids; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D020935:MAP Kinase Signaling System; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D018384:Oxidative Stress; D010766:Phosphorylation; D017382:Reactive Oxygen Species / Q000378:metabolism; D013314:Stress, Mechanical; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
D005419:Flavonoids; D017382:Reactive Oxygen Species; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; D048051:p38 Mitogen-Activated Protein Kinases; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one