on July 31, 2003
Published online before print July 31, 2003, doi: 10.1161/01.ATV.0000089012.73180.63
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Submitted on January 7, 2003
Accepted on May 6, 2003
Ginkgo biloba Extract Inhibits Tumor Necrosis Factor-
-Induced Reactive Oxygen Species Generation, Transcription Factor Activation, and Cell Adhesion Molecule Expression in Human Aortic Endothelial Cells
Jaw-Wen Chen *;
From National Yang-Ming University School of Medicine, Cardiovascular Research Center and Division of Cardiology, Department of Medicine, Taipei Veterans General Hospital, Taiwan, Republic of China.
* To whom correspondence should be addressed. E-mail: jwchen{at}vghtpe.gov.tw.
Objective--This study was conducted to examination whether Ginkgo biloba extract (GBE), a Chinese herb with antioxidant activity, could reduce cytokine-induced monocyte/human aortic endothelial cell (HAEC) interaction, a pivotal early event in atherogenesis.
Methods and Results--Pretreatment of HAECs with GBE (50 and 100 µg/mL for 18 hours) significantly suppressed cellular binding between the human monocytic cell line U937 and tumor necrosis factor-
(TNF-)-stimulated HAECs by using in vitro binding assay (68.7% and 60.1% inhibitions, respectively). Cell enzyme-linked immunosorbent assay and immunoblot analysis showed that GBE (50 µg/mL for 18 hours) significantly attenuated TNF--induced cell surface and total protein expression of vascular cellular adhesion molecule-1 and intracellular adhesion molecule-1 (63.5% and 69.2%, respectively; P<0.05). However, pretreatment with probucol (5 µmol/L for 18 hours) reduced the expression of vascular cellular adhesion molecule-1 but not intracellular adhesion molecule-1. Preincubation of HAECs with GBE or probucol significantly reduced intracellular reactive oxygen species formation induced by TNF- (76.8% and 68.2% inhibitions, respectively; P<0.05). Electrophoretic mobility shift assay demonstrated that both GBE and probucol inhibited transcription factor nuclear factor-
B activation in TNF--stimulated HAECs (55.2% and 65.6% inhibitions, respectively) but only GBE could inhibit the TNF--stimulated activator protein 1 activation (45.1% inhibition, P<0.05).
Conclusions--GBE could reduce cytokine-stimulated endothelial adhesiveness by downregulating intracellular reactive oxygen species formation, nuclear factor-B and activator protein 1 activation, and adhesion molecule expression in HAECs, supporting the notion that the natural compound Ginkgo biloba may have potential implications in clinical atherosclerosis disease.
Key words: activator protein 1 • cell adhesion molecule • Ginkgo biloba • human aortic endothelial cells • nuclear factor-
B
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