Cyclosporin A inhibits the adhesion of neutrophil with ECV-304 induced by hypoxia/reoxygenation via ROS-Cyclophilin A-ERK1/2 pathway.
- Author:
Si-Gui ZHOU
1
;
Li-Peng XU
;
Duan-Fang LIAO
;
Xiao-Yong LEI
;
Feng-Xiang YAN
;
Bing-Yang ZHU
Author Information
1. Institute of Pharmacy and Pharmacology, Nanhua University, Hengyang, Hunan 421001, China.
- Publication Type:Journal Article
- MeSH:
Cell Adhesion;
Cell Hypoxia;
Cells, Cultured;
Cyclophilins;
biosynthesis;
genetics;
Cyclosporine;
pharmacology;
Endothelium, Vascular;
cytology;
Humans;
Intercellular Adhesion Molecule-1;
biosynthesis;
genetics;
Mitogen-Activated Protein Kinase 1;
metabolism;
Mitogen-Activated Protein Kinase 3;
metabolism;
Neutrophils;
cytology;
Reactive Oxygen Species;
metabolism;
Reperfusion Injury;
physiopathology;
Signal Transduction;
Umbilical Veins;
cytology
- From:
Acta Physiologica Sinica
2004;56(3):313-320
- CountryChina
- Language:Chinese
-
Abstract:
To investigate the inhibition of cyclosporin A (CsA) on neutrophil adhesion to human umbilical vein endothelial cells (HUVECs, ECV-304) induced by hypoxia/reoxygenation and further explore its mechanism, a 1 h hypoxia/4 h reoxygenation model was reproduced using ECV-304. The adhesion rate of neutrophils to ECV-304 was determined by measuring the activity of endogenous hexosaminidase. The expression of endothelial cell adhesion molecules of E-selectin and ICAM-1 was measured by flow cytometry. The expression of cyclophilin A (CyPA) and the activation of ERK1/2 was compared among experimental groups by Western blot. The content of reactive oxygen species (ROS) was measured by Fenton reaction. After being stimulated with 1 h hypoxia/4 h reoxygenation, ECV-304 showed an enhanced neutrophil adhensiveness in association with an increased surface expression of E-selectin and ICAM-1. In parallel, the content of ROS was also increased. These effects were significantly suppressed by the addition of CsA. Most importantly, the expression of CyPA was significantly increased following 1 h hypoxia/4 h reoxygenation, which was accompanied with an increased activation of ERK1/2. Treatment with CyPA inhibitor CsA and CyPA antisense oligonucleotides significantly inhibited the activation of ERK1/2 and decreased the adhesion of neutrophils to ECV-304. The specific ERK1/2 inhibitor PD98059 caused an inhibition of neutrophil adhesion to hypoxia/reoxygenation-stimulated ECV-304. Our data confirm that CsA inhibits neutrophil adhesion to hypoxia/reoxygenation stimulated ECV-304 by a mechanism involving inhibition of the signal transduction of ROS, CyPA and ERK1/2.
