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Vol. 293, Issue 3, 982-988, June 2000
Department of Pharmacology, Shanghai Institute of Materia Medica,
Chinese Academy of Sciences, Shanghai, China
Although clinical studies have demonstrated that EGb 761, a standard
extract of Ginkgo biloba, was effective in
mild-to-moderate dementia of the Alzheimer's disease patients, the
mechanism underlying its neuroprotective effect remains unclear. In
this study, effects of bilobalide, the main constituent of the
nonflavone fraction of EGb 761, on reactive oxygen species
(ROS)-induced apoptosis in PC12 cells was studied. Exposure of cells to
xanthine (100 µM)/xanthine oxidase (150 mU/ml) (ROS producer)
resulted in a characteristic DNA fragmentation and an increase in the
apoptosis rate. When p53, c-Myc, Bcl-2, Bcl-xL, and Bax
were measured by flow cytometry and the activities of caspase-1- and
caspase-3-like protease determined with Ac-YVAD-AMC or Ac-DEVD-AMC as
substrates, the profile of ROS-induced changes in these apoptosis
regulatory and effector proteins suggests that elevation of c-Myc, p53,
and Bax and activation of caspase-3 play an important role in the apoptosis. When cells were treated with ROS and bilobalide (25-100 µM) simultaneously, a dose-dependent reduction in the apoptotic rate
was found. The percentage of cells with positive staining for c-Myc and
p53 decreased from 27.8 and 50.1% to 16.7 and 23.2%, respectively,
when bilobalide (25 µM) was present. Bilobalide also reduced
ROS-induced elevation of Bax and activation of caspase-3 effectively.
Our results provide the first direct evidence that bilobalide can
protect neurons against oxidative stress. Bilobalide may block the
apoptosis in the early stage and then attenuate the elevation of c-Myc,
p53, and Bax and activation of caspase-3 in cells.
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