TGF-beta(1) down-regulates inflammatory cytokine-induced VCAM-1 expression in cultured human glomerular endothelial cells.
BACKGROUND
Endothelial cells are active participants in the processes controlling coagulation, inflammation and the immune response. Variations are recognized between endothelia isolated from different vascular beds as well as from different species. Though transforming growth factor-beta(1) (TGF-beta(1)) has been known to have an anti-inflammatory action, little is known about its effect on expression of cellular adhesion molecules during the inflammatory process in human glomerular endothelial cells. The aim of this study was to determine the effect of TGF-beta(1) on the inflammatory cytokine-induced expression of vascular cell adhesion molecule-1 (VCAM-1) in cultured human glomerular endothelial cells.


METHODS
The culture of human glomerular endothelial cells was established using the normal portion of nephrectomized renal tissues and identified by factor VIII staining and cellular uptake of fluorescent-labelled acetylated low-density lipoprotein (LDL). The endothelial cells were stimulated by interleukin-1beta (IL-1beta), tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) with or without TGF-beta(1). Cellular expression of VCAM-1 was measured by enzyme-linked immunosorbent assay (ELISA) and flow cytometry, and VCAM-1 mRNA was measured by Northern blot analysis.


RESULTS
TGF-beta(1) (1, 10 and 25 ng/ml) blunted IL-1beta- (5 ng/ml) induced VCAM-1 expression significantly (OD=1.08+/-0.14, 1. 10+/-1.16 and 1.05+/-0.14 vs IL-1beta=1.97+/-0.29, n=6, P<0.05) in ELISA. The addition of TGF-beta(1) (1, 10 and 25 ng/ml) also suppressed TNF-alpha- (10 ng/ml) induced VCAM-1 expression (OD=1. 14+/-0.15, 1.17+/-0.17 and 1.18+/-0.16 vs TNF-alpha=1.96+/-0.26, n=6, P<0.05). The same results were obtained by flow cytometry. TGF-beta(1) (10 ng/ml) inhibited both IL-1beta- (5 ng/ml) and TNF-alpha-(10 ng/ml) induced expression of VCAM-1 (MFI: IL-1beta=90. 8+/- 17.6, IL-1beta+TGF-beta(1)=37.8+/-14.9, TNF-alpha=113.6+/- 12.4, TNF-alpha+TGF-beta(1)=64.3+/-13.8, mean+/-SD, n=3, P<0.05). By Northern blot analysis, TGF-beta(1) (10 ng/ml) significantly suppressed the stimulatory effect of IL-1beta and TNF-alpha.


CONCLUSIONS
These results show that TGF-beta(1) down-regulates the inflammatory cytokine-induced expression of VCAM-1 in human glomerular endothelial cells, which could be a novel mechanism for the anti-inflammatory action of TGF-beta(1) during the inflammatory processes in human glomerular diseases.
Park
S K
SK
Departments of Internal Medicine, Urology and Biochemistry, Asan Medical Center, College of Medicine, University of Ulsan, Seoul, Korea.
Yang
W S
WS
Lee
S K
SK
Ahn
H
H
Park
J S
JS
Hwang
O
O
Lee
J D
JD
D002478:Cells, Cultured; D016207:Cytokines / Q000494:pharmacology*; D003907:Dexamethasone / Q000494:pharmacology; D004727:Endothelium / Q000166:cytology / Q000378:metabolism; D005938:Glucocorticoids / Q000494:pharmacology; D006801:Humans; D018836:Inflammation Mediators / Q000494:pharmacology*; D007678:Kidney Glomerulus / Q000166:cytology / Q000378:metabolism*; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism*
D016207:Cytokines; D005938:Glucocorticoids; D018836:Inflammation Mediators; D016212:Transforming Growth Factor beta; D019010:Vascular Cell Adhesion Molecule-1; D003907:Dexamethasone