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Journal of Chinese Integrative Medicine ›› 2004, Vol. 2 ›› Issue (5): 367-371.doi: 10.3736/jcim20040517

• Original Experimental Research • Previous Articles     Next Articles

Effect of Bugu Mixture on all-trans retinoic acid-induced apoptosis of bone marrow stromal cells

Nan Li1(), He-ming Wang1, Xu Lin2, Liang-pu Zheng3, Lin Shen4, Yi-ping Lin5, Lin-ying Zhou6, Xiu-rong Zhong6, Fang Lin7   

  1. 1. Department of Orthopedics and Traumatology, Fujian Traditional Chinese Medical College, Fuzhou, Fujian Province 350003, China
    2. Department of Orthopedics and Traumatology, Fujian Traditional Chinese Medical College, Fuzhou, Fujian Province 350003, China
    3. Molecular Medicine Center, Fujian Medical University, Fuzhou, Fujian Province 350004, China
    4. Medical Science Experiment Center, Fujian Traditional Chinese Medical College, Fuzhou, Fujian Province 350003, China
    5. Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province 430022, China
    6. Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province 430022, China
    7. Department of Traditional Chinese Basic Medicine, Fujian Academy of Traditional Chinese Medicine, Fuzhou, Fujian Province 350003, China
  • Received:2003-11-05 Online:2004-09-20 Published:2018-10-20

Objective: We used the SD rat's bone marrow stromal cells (BMSCs) cultured in vitro to observe the effects of Bugu Mixture on the apoptosis and to explore the molecular biologic mechanism of the treatment of osteoporosis with Bugu Mixture.Methods: BMSCs were separated from the bones of the extremities of SD rats in vitro. The morphologic changes, the apoptosis cell cycles, the mitochondrion membrane potential changes, and the Bcl-2 and Bax gene expression were observed, and the effects of Bugu Mixture on the course of cell apoptosis were evaluated.Results: The earlier use of Bugu Mixture could decrease the cells blocked in G0/G1 phase, and promote their synthesis of DNA in S phase. The expression of Bcl-2 was higher in the Bugu Mixture group than that in the all-trans retinoic acid(ATRA) induced group, and the expression of Bax was lower in the Bugu Mixture group than that in the ATRA induced group. The mitochondrion membrane potential descended significantly in the Bugu Mixture group than that in the ATRA induced group.Conclusion: The mechanism of the treatment of osteoporosis with Bugu Mixture is that the earlier use of Bugu Mixture can decrease the amount of apoptostic cells induced by ATRA, thus promoting the cell mitosis and restraining the apoptosis. It can also act as a protector to Bcl-2 located on the mitochondrion membrane. By preventing the transferring of the Bax protein from cell-plasm to mitochondrion membrane, it takes the advantage of Bcl-2 in forming Bcl-2/Bax homodimer so as to prevent the opening of the permeability transition pore to avoid the changing of mitochondrion membrane potential and the destruction of biosynthesis caused by the mitochondrion release of apoptosis inducing factors and to reach the objective of restraining apoptosis.

Key words: osteoporosis, bone marrow cells, apoptosis, genes, Bcl-2, mitochondria, membrane potentials, animals, laboratory

CLC Number: 

  • R289.5

Fig 1

Morphosis of BMSC apoptosis induced by 10-4 mol/L ATRA (Dyeing by AO-EB, fluorescence microscope, ×1 000)"

Fig 2

Bcl-2 dyeing in control group (×100)"

Fig 3

Bax dyeing in ATRA group (×100)"

Fig 4

Result of FCM detection A: Control group; B: ATRA induced group; C: ATRA and Bugu Mixture group; D: Bugu Mixture (3 d) and ATRA group"

Tab 1

Effect of different treatment on BMSC cycle and apoptosis rate ($\bar{x}$±s, %)"

Group n G1 G2 S Apoptosis rate
Control group 6 57.27±2.38 11.17±3.32 31.61±2.15 0
ATRA induced group 6 82.76±4.90** 10.98±4.49 6.26±1.26** 37.07±2.81
ATRA and Bugu Mixture group 6 68.15±3.87**△△ 21.29±7.51 11.56±3.47**△△ 15.21±3.87△△
Bugu Mixture (3d) and ATRA group 6 60.60±7.15 20.18±3.81 19.23±3.70**▲▲ 7.36±1.68△△▲▲

Tab 2

Effect of different treatment on BMSC ΔΨm ($\bar{x}$±s, %)"

Group n FITC positive rate of BMSC
24 h 48 h
Control group 5 0.58±0.16 0.65±0.38
ATRA induced group 5 6.91±0.45** 62.79±1.48**
Bugu Mixture group 5 4.06±0.46**△△ 24.40±1.64**△△

Tab 3

Relative expression quantity of Bax and Bcl-2 mRNA at different time in different groups"

Group Time Bcl-2 Bax Bcl-2/Bax
Control group
0.227 4 0.100 0 2.274 0
ATRA induced group
1st day 0.082 7 0.332 3 0.248 9
2nd day 0.059 3 0.621 6 0.095 4
3rd day 0.039 1 0.632 2 0.061 9
ATRA and Bugu Mixture group
1st day 0.136 7 0.118 0 1.158 5
2nd day 0.107 5 0.279 6 0.384 5
3rd day 0.150 0 0.205 9 0.728 5
Bugu Mixture (3d) and ATRA group
1st day 0.219 6 0.074 5 2.947 7
2nd day 0.150 4 0.540 9 0.278 1
3rd day 0.124 1 0.257 1 0.482 7
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