Eotaxin increases monolayer permeability of human coronary artery endothelial cells.
OBJECTIVE
The objective of this study was to determine the effects and molecular mechanisms of eotaxin, a newly discovered chemokine (CCL11), on endothelial permeability in the human coronary artery endothelial cells (HCAECs).


RESULTS
Cells were treated with eotaxin, and the monolayer permeability was studied by using a costar transwell system with a Texas Red-labeled dextran tracer. Eotaxin significantly increased monolayer permeability in a concentration-dependent manner. In addition, eotaxin treatment significantly decreased the mRNA and protein levels of endothelial junction molecules including zonula occludens-1 (ZO-1), occludin, and claudin-1 in a concentration-dependent manner as determined by real-time RT-PCR and Western blot analysis, respectively. Increased oxidative stress was observed in eotaxin-treated HCAECs by analysis of cellular glutathione levels. Furthermore, eotaxin treatment substantially activated the phosphorylation of MAPK p38. HCAECs expressed CCR3. Consequently, antioxidants (ginkgolide B and MnTBAP), specific p38 inhibitor SB203580, and anti-CCR3 antibody effectively blocked the eotaxin-induced permeability increase in HCAECs. Eotaxin also increased the phosphorylation of Stat3 and nuclear translocation of NF-kappaB in HCAECs.


CONCLUSIONS
Eotaxin increases vascular permeability through CCR3, the downregulation of tight junction proteins, increase of oxidative stress, and activation of MAPK p38, Stat3, and NF-kB pathways in HCAECs.
Molecular Surgeon Research Center, Division of Vascular Surgery and Endovascular Therapy, Michael E. DeBakey Department of Surgery, Baylor College of Medicine, Houston, TX 77030, USA.
Md Saha
MS
Jamaluddin
Xinwen
X
Wang
Hao
H
Wang
Cubas
C
Rafael
Qizhi
Q
Yao
Changyi
C
Chen
D002199:Capillary Permeability / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D054413:Chemokine CCL11 / Q000494:pharmacology* / Q000502:physiology; D062445:Claudin-1; D003331:Coronary Vessels / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D042783:Endothelial Cells / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D062793:Occludin; D018384:Oxidative Stress / Q000187:drug effects; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D054397:Receptors, CCR3 / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000378:metabolism; D062826:Zonula Occludens-1 Protein; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
C515456:CCL11 protein, human; C515350:CCR3 protein, human; C568108:CLDN1 protein, human; D054413:Chemokine CCL11; D062445:Claudin-1; D008565:Membrane Proteins; D016328:NF-kappa B; C568985:OCLN protein, human; D062793:Occludin; D010750:Phosphoproteins; D012333:RNA, Messenger; D054397:Receptors, CCR3; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; C569042:TJP1 protein, human; D062826:Zonula Occludens-1 Protein; D048051:p38 Mitogen-Activated Protein Kinases