Nephrol Dial Transplant (2000) 15: 596-604
© 2000 European Renal Association-European Dialysis and Transplant Association
TGF-ß1 down-regulates inflammatory cytokine-induced VCAM-1 expression in cultured human glomerular endothelial cells
Departments of Internal Medicine, Urology and Biochemistry, Asan Medical Center, College of Medicine, University of Ulsan, Seoul, Korea
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-ß1 (TGF-ß1) 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-ß1 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-1ß (IL-1ß), tumour necrosis factor-
(TNF-) and interferon-
(IFN-) with or without TGF-ß1. 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-ß1 (1, 10 and 25 ng/ml) blunted IL-1ß- (5 ng/ml) induced VCAM-1 expression significantly (OD=1.08±0.14, 1.10±1.16 and 1.05±0.14 vs IL-1ß=1.97±0.29, n=6, P<0.05) in ELISA. The addition of TGF-ß1 (1, 10 and 25 ng/ml) also suppressed TNF-- (10 ng/ml) induced VCAM-1 expression (OD=1.14±0.15, 1.17±0.17 and 1.18±0.16 vs TNF-=1.96±0.26, n=6, P<0.05). The same results were obtained by flow cytometry. TGF-ß1 (10 ng/ml) inhibited both IL-1ß- (5 ng/ml) and TNF--(10 ng/ml) induced expression of VCAM-1 (MFI: IL-1ß=90.8± 17.6, IL-1ß+TGF-ß1=37.8±14.9, TNF-=113.6± 12.4, TNF-+TGF-ß1=64.3±13.8, mean±SD, n=3, P<0.05). By Northern blot analysis, TGF-ß1 (10 ng/ml) significantly suppressed the stimulatory effect of IL-1ß and TNF-.
Conclusions. These results show that TGF-ß1 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-ß1 during the inflammatory processes in human glomerular diseases.
Keywords: cytokines; culture; glomerular endothelial cells; TGF-ß1; VCAM-1
Correspondence and offprint requests to: Su-Kil Park, MD, PhD, Department of Internal Medicine, Asan Medical Center, College of Medicine, University of Ulsan, Song-Pa, PO Box 145, Seoul 138-736, Korea.
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