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M309114200v1
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Papers In Press, published online ahead of print November 24, 2003
J. Biol. Chem, 10.1074/jbc.M309114200
Submitted on August 18, 2003
Revised on November 16, 2003
Accepted on November 24, 2003

A constituent of green tea, Epigallocatechin-3-gallate, activates endothelial nitric oxide synthase by a PI3K-, PKA-, and Akt-dependent pathway, and leads to endothelial-dependent vasorelaxation

Mario Lorenz, Silja Wessler, Elena Follmann, Wanda Michaelis, Thomas Düsterhöft, Gert Baumann, Karl Stangl, and Verena Stangl

Med. Department, Cardiology, Charité, Humboldt-University, CCM, Berlin 10117

Corresponding Author: verena.stangl{at}charite.de

Epidemiological studies suggest that tea catechins may reduce the risk of cardiovascular disease, but the mechanisms of benefit have not been determined. The objective of the present study was to investigate the effects of epigallocatechin-3-gallate (EGCG), the major constituent of green tea, on vasorelaxation and on eNOS expression and activity in endothelial cells. EGCG (1-50 µM) induced dose-dependent vasodilation in rat aortic rings. Vasodilation was abolished by pretreatment with L-NAME. In bovine aortic endothelial cells (BAEC), EGCG dose-dependently increased eNOS activity after 15 minutes. Treatment with EGCG induced a sustained activation of Akt, ERK1/2, and eNOS Ser1179 phosphorylation. Inhibition of ERK1/2 had no influence on eNOS activity or Ser1179 phosphorylation. Simultaneous treatment of cells with selective inhibitors for PKA and Akt completely prevented the increase in eNOS activity by EGCG after 15 minutes, indicating that both kinases act in concert. Specific PI3K inhibitors yielded identical results. Akt inhibition prevented eNOS Ser1179 phosphorylation whereas inhibition of PKA did not influence Akt and eNOS Ser1179 phosphorylation. Pretreatment of endothelial cells with EGCG for 4 hours markedly enhanced the increase in eNOS activity stimulated by Ca-ionomycin, suggesting that Akt accounts for prolonged eNOS activation. Treatment of cells for 72 hours with EGCG did not change eNOS protein levels. Our results indicate that EGCG-induced endothelium-dependent vasodilation is primarily based on rapid activation of eNOS by a PI3K-, PKA-, and Akt-dependent increase in eNOS activity, independently of an altered eNOS protein content.


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