Panax notoginseng saponins protect rabbit bone marrow stromal cells from hydrogen peroxide-induced apoptosis.
OBJECTIVE
To investigate the effects of Panax notoginseng saponins (PNSs) on hydrogen peroxide-induced apoptosis in rabbit bone marrow stromal cells (BMSCs).


METHODS
BMSCs were isolated from 2-month-old New Zealand rabbits and cultured with different doses of PNSs to determine the most effective dose of PNSs by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay and alkaline phosphatase (ALP) assay. The most effective dose of PNSs was used in subsequent experiments. Apoptosis of BMSCs was induced by hydrogen peroxide (100 micromol/L). BMSCs in PNSs group were also pretreated with PNSs before hydrogen peroxide exposure. Reactive oxygen species (ROSs) levels were measured by using 2',7'-dichlorodihydrofluorescein diacetate. Apoptosis rate of BMSCs was observed by flow cytometry after staining with Annexin V-fluorescein isothiocyanate/propidium iodide. The protein expression of Bax in BMSCs was analyzed by Western blotting. Activity of caspase-3 was measured by spectrofluorometry.


RESULTS
The most effective dose of PNSs was 0.1g/L. PNSs at dose of 0.1g/L markedly reversed the augmentation of ROS level, decreased the apoptosis rate of, and the Bax expression and activity of caspase-3 in BMSCs treated with hydrogen peroxide (P<0.01).


CONCLUSIONS
PNSs can protect cultured rabbit BMSCs from hydrogen peroxide-induced apoptosis by decreasing oxidative stress, Bax expression and caspase-3 activity.
Department of Orthopedics, the Second Affiliated Hospital, Xi'an Jiaotong University College of Medicine, Xi'an 710004, Shaanxi Province, China.
Hui
H
Qiang
Kun-zheng
KZ
Wang
Zhi-bin
ZB
Shi
Li-hong
LH
Fan
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001854:Bone Marrow Cells / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D036145:Ginsenosides / Q000494:pharmacology*; D006861:Hydrogen Peroxide / Q000009:adverse effects*; D018384:Oxidative Stress / Q000187:drug effects*; D053723:Panax notoginseng / Q000737:chemistry*; D011817:Rabbits; D017154:Stromal Cells / Q000187:drug effects; D051028:bcl-2-Associated X Protein / Q000378:metabolism
D036145:Ginsenosides; D051028:bcl-2-Associated X Protein; D006861:Hydrogen Peroxide; D053148:Caspase 3