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Journal of Chinese Integrative Medicine ›› 2010, Vol. 8 ›› Issue (2): 131-137.doi: 10.3736/jcim20100207

• Original Experimental Research • Previous Articles     Next Articles

Panax notoginseng saponins protect rabbit bone marrow stromal cells from hydrogen peroxide-induced apoptosis

 Hui Qianga,Kun-zheng Wanga, Chen Zhanga, Zhi-bin Shia, Li-hong Fana   

  1. a Department of Orthopedics, the Second Affiliated Hospital, Xi5an Jiaotong University College of Medicine, Xi’an 710004, Shaanxi Province, China;
    b Department of Orthopedics,Shaanxi Provincial People’s Hospital,Xi’an 710068,Shaanxi Province,China;
  • Received:2009-09-22 Accepted:2010-01-10 Online:2010-02-20 Published:2010-02-15

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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 μmol/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 0.1 g/L. PNSs at dose of 0.1 g/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.

Key words: Panax notoginseng , Hydrogen peroxide, Bone marrow stromal cells, Apoptosis, Rabbits

Figure 1

Effects of different doses of PNSs on BMSCsA: Growth curves of BMSCs detected by using the MTT assay; B: ALP activity of BMSCs. *P<0.05, vs 0 g/L group."

Figure 2

Effects of PNSs on intracellular ROS level in BMSCs A: Control group; B: H2O2 group; C: PNSs (0.1 g/L) group; **P<0.01, vs control group; ΔΔP<0.01, vs H2O2 group."

Figure 3

Effects of PNSs on BMSCs apoptosis assessed by flow cytometryA: Control group; B: H2O2 group; C: PNSs (0.1 g/L) group. **P<0.01, vs control group; ΔΔP<0.01, vs H2O2 group."

Figure 4

Effects of PNSs on bax protein expression in BMSCs A: Control group; B: H2O2 group; C: PNSs (0.1 g/L) group. a: Bax expression analyzed by Western blotting; b: Comparison of Bax relative expression. **P<0.01, vs control group; ΔΔP<0.01, vs H2O2 group."

Figure 5

Effects of PNSs on caspase-3 activity in BMSCs A: Control group; B: H2O2 group; C: PNSs (0.1 g/L) group. **P<0.01, vs control group; ΔΔP<0.01, vs H2O2 group."

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