Arsenite induces endothelial cell permeability increase through a reactive oxygen species-vascular endothelial growth factor pathway.
As a potent environmental oxidative stressor, arsenic exposure has been reported to exacerbate cardiovascular diseases and increase vascular endothelial cell monolayer permeability. However, the underlying mechanism of this effect is not well understood. In this paper, we test our hypothesis that reactive oxygen species (ROS)-induced vascular endothelial growth factor (VEGF) expression may play an important role in an arsenic-caused increase of endothelial cell monolayer permeability. The mouse brain vascular endothelial cell bEnd3 monolayer was exposed to arsenite for 1, 3, and 6 days. The monolayer permeability, VEGF protein release, and ROS generation were determined. In addition, VE-cadherin and zonula occludens-1 (ZO-1), two membrane structure proteins, were immunostained to elucidate the effects of arsenite on the cell-cell junction. The roles of ROS and VEGF in arsenite-induced permeability was determined by inhibiting ROS with antioxidants and immuno-depleting VEGF with a VEGF antibody. We observed that arsenite increased bEnd3 monolayer permeability, elevated the production of cellular ROS, and increased VEGF release. VE-cadherin and ZO-1 disruptions were also found in cells treated with arsenite. Furthermore, both antioxidant (N-acetyl cysteine and tempol) and the VEGF antibody treatments significantly lowered the arsenite-induced permeability of the bEnd3 monolayer as well as VEGF expression. VE-cadherin and ZO-1 disruptions were also diminished by N-acetyl cysteine and the VEGF antibody. Our data suggest that the increase in VEGF expression caused by ROS may play an important role in the arsenite-induced increase in endothelial cell permeability.
Department of Pharmacology and Toxicology, School of Pharmacy, University of Kansas, Lawrence, Kansas 66045, USA.
Lingzhi
L
Bao
Honglian
H
Shi
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D018053:Arsenites / Q000737:chemistry / Q000633:toxicity*; D015820:Cadherins / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects*; D042783:Endothelial Cells / Q000378:metabolism; D004785:Environmental Pollutants / Q000737:chemistry / Q000633:toxicity*; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D010750:Phosphoproteins / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013997:Time Factors; D042442:Vascular Endothelial Growth Factors / Q000276:immunology / Q000378:metabolism*; D062826:Zonula Occludens-1 Protein
D000906:Antibodies; D000975:Antioxidants; D018053:Arsenites; D015820:Cadherins; D004785:Environmental Pollutants; D008565:Membrane Proteins; D010750:Phosphoproteins; D017382:Reactive Oxygen Species; C569043:Tjp1 protein, mouse; D042442:Vascular Endothelial Growth Factors; D062826:Zonula Occludens-1 Protein; C015001:arsenite