Effect of 11, 12-epoxyeicosatrienoic acids on hypoxia/reoxygenation injury in the human umbilical vein endothelial cells.
- Author:
Xiao-wei QIU
1
;
Wen WANG
;
Dong-qiao JIANG
;
Hong-xia WANG
;
Li YAN
;
Xiao-yan WANG
;
Li-quan MA
;
Ling-Qiao LU
;
Chao-shu TANG
;
Li-ke ZHANG
Author Information
- Publication Type:Journal Article
- MeSH: 8,11,14-Eicosatrienoic Acid; analogs & derivatives; pharmacology; Cell Hypoxia; drug effects; physiology; Cell Survival; Cells, Cultured; Endothelial Cells; drug effects; Humans; L-Lactate Dehydrogenase; metabolism; Malondialdehyde; metabolism; Mitogen-Activated Protein Kinase 3; biosynthesis; Nitric Oxide Synthase Type III; biosynthesis; Reperfusion Injury; prevention & control; Superoxide Dismutase; metabolism; Umbilical Veins; cytology
- From: Acta Academiae Medicinae Sinicae 2006;28(6):803-807
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the effects of 11, 12-epoxyeicosatrienoic acids (11, 12-EET) on the degree of hypoxia/reoxygenation injury in human umbilical vein endothelial cells ( HUVECs), and reveal the possible pathway of EET on protection.
METHODSPrimary cultured HUVECs were randomly divided into control group, hypoxia/reoxygenation group, 11, 12-EET control group, 11, 12- EET hypoxia/reoxygenation group, inhibition of extracellular signal-regulated kinase (ERKI/2) group, and inhibition of nitric oxide synthase (NOS) group. Hypoxia/reoxygenation injury model in HUVECs was established by exposure to hypoxia (2% O2, 5% CO2 and 93% N2) for 3 hours, followed by reoxygenation (95% air and 5% CO2) for 1 hour. The evaluation of the endothelial cells were made by immunohistochemistry. The cell viability was monitored by MTT assay. Colorimetry method was used to assay the lactate dehydrogenase (LDH) , malondialdehyde (MDA) and activity of superoxide dismutase (SOD) in culture medium. Western blot was used to detect the expressions of endothelial nitric oxide synthase (eNOS) and phosphorylated ERK1/2 in HUVECs.
RESULTS11, 12-EET caused minor injury in normal oxygen incubated HUVECs; however, in hypoxia/reoxygenation HUVECs, it raised the cell viability markedly, decreased the LDH release and MDA content, and increased the activity of SOD and the expressions of eNOS and phosphorylated ERK1/2.
CONCLUSIONS11, 12-EET may prevent against endothelial cell hypoxia/reoxygenation injury. The mechanism may be related to the increased activity of SOD, elimination of oxygen-derived free radicals, and reduction of eNOS and phosphorylated ERK1/2 lesion caused by hypoxia/reoxygenation.