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Panax notoginseng saponins protect rabbit bone marrow stromal cells from hydrogen peroxide-induced apoptosis
Hui Qiang1, Kun-zheng Wang, Zhi-bin Shi
1Department of Orthopedics, the Second Affiliated Hospital, Xi'an Jiaotong University College of Medicine, Xi'an 710004, Shaanxi Province, China.
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).
Conclusion:
PNSs can protect cultured rabbit BMSCs from hydrogen peroxide-induced apoptosis by decreasing oxidative stress, Bax expression and caspase-3 activity.