Expression and regulation of vascular cell adhesion molecule-1 in human umbilical vein endothelial cells induced by sera from severely burned patients.
OBJECTIVE
The purpose of this study was to determine the expression and regulation of vascular cell adhesion molecule (VCAM)-1 in human umbilical vein endothelial cells (HUVECs) induced by sera from severely burned patients.


METHODS
Controlled laboratory study.


METHODS
Research laboratory in a university hospital.


METHODS
HUVECs.


METHODS
HUVECs were incubated with serum from eight healthy controls and eight patients with thermal injuries of >50% total body surface area. The experiment was repeated after pretreatment with pyrrolidine dithiocarbamate, an inhibitory effect on nuclear factor-kappaB activation, SB203580, a specific p38 mitogen-activated protein kinase inhibitor, and PD98059, a mitogen-activated protein/extracellular signal-regulated kinase inhibitor.


RESULTS
Protein and messenger RNA expression of VCAM-1 was measured by flow cytometry and reverse transcription-polymerase chain reaction respectively. Soluble VCAM-1 level in HUVECs culture supernatants was measured by enzyme-linked immunosorbent assay. Sera from severely burned patients showed a stimulatory effect on VCAM-1 messenger RNA levels and an increased VCAM-1 expression on the endothelial cell surfaces. The soluble form of VCAM-1 molecules was also elevated by the stimulation of burn sera. In vitro peripheral blood mononuclear leukocytes adherence to HUVECs incubated with burn sera was significantly increased compared with those incubated with control sera. Finally, these events were significantly inhibited by pretreatment with antioxidants pyrrolidine dithiocarbamate or SB203580, whereas PD98059 had no significant effect.


CONCLUSIONS
These findings suggest that sera from severely burned patients induced up-regulation of VCAM-1 expressions in HUVECs, and this process might be largely dependent on oxidant-mediated nuclear factor-kappaB activation and p38 mitogen-activated protein kinase pathways.
Chen
Xu-Lin
XL
Burns Center, Changhai Hospital, Second Military University, Shanghai, People's Republic of China.
Xia
Zhao-Fan
ZF
Wei
Duo
D
Liao
He-Gen
HG
Ben
Dao-Feng
DF
Wang
Guang-Qing
GQ
D000328:Adult; D015415:Biomarkers / Q000097:blood; D001707:Biopsy, Needle; D002056:Burns / Q000097:blood* / Q000473:pathology; D016022:Case-Control Studies; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D015601:Injury Severity Score; D008297:Male; D016328:NF-kappa B / Q000502:physiology*; D011379:Prognosis; D012016:Reference Values; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014471:Umbilical Veins; D015854:Up-Regulation; D019010:Vascular Cell Adhesion Molecule-1 / Q000032:analysis*
D015415:Biomarkers; D016328:NF-kappa B; D019010:Vascular Cell Adhesion Molecule-1