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Vol. 60, Issue 3, 528-533, September 2001
)-Epigallocatechin-3-Gallate Blocks
Nuclear Factor-
B Activation by Inhibiting IB Kinase Activity in
the Intestinal Epithelial Cell Line IEC-6
Graduate Program in Nutritional Sciences (F.Y., S.B., W.J.S.D.V.,
C.J.M., G.W.V.) and Department of Internal Medicine (W.J.S.D.V.),
University of Kentucky and Veterans Administration Medical Center,
Lexington, Kentucky; and Departments of Internal Medicine (H.S.O.,
S.B., C.J.M., G.W.V.) and Pharmacology and Toxicology (S.B., C.J.M.),
University of Louisville and Veterans Administration Medical Center,
Louisville, Kentucky
The I
B kinase complex (IKK) mediates activation of the
transcription factor nuclear factor-B (NF-B). We previously
showed that green tea polyphenols inhibited endotoxin-mediated tumor necrosis factor-
(TNF) production by blocking NF-B activation. In this study, we evaluated whether green tea polyphenols inhibit NF-B by blocking IKK activity. We assessed IKK activity by detecting changes in phosphorylation of an IB-glutathione
S-transferase (GST) fusion protein. IEC-6 cells
pretreated with an extract of green tea polyphenols (GrTPs; 0-0.4
mg/ml) had diminished TNF-induced IKK and NF-B activity. Of the
various GrTPs, (
)-epigallocatechin-3-gallate (EGCG) was the most
potent inhibitor. We next examined whether EGCG inhibited activated
IKK. In cytosolic extracts of TNF-stimulated cells, EGCG inhibited
phosphorylation of IB-GST (IC50 > 18 µM) consistent with inhibition of IKK activity. Using other polyphenols, we
showed that the gallate group was essential for inhibition, and
antioxidants were ineffective in blocking activated IKK. Importantly, EGCG decreased IKK activity in cytosolic extracts of NIK transiently transfected cells. This latter finding showed that our findings were
not related to nonspecific kinase activity. In conclusion, EGCG is an
effective inhibitor of IKK activity. This may explain, at least in
part, some of the reported anti-inflammatory and anticancer effects of
green tea.
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