| Objective According to the theory of traditional Chinese medicine, Bu Shen Zhunag Gu, Qiang Gu Kang Shu capsules is prepared and observed the effect in ovariectomized rats on bone mineral density (BMD), bone histomorphometry and microstructure, and expression of OPG and RANKL in bone marrow stroma cells, to evaluate whether or not Qiang Gu Kang Shu capsules can prevent osteoporosis from estrogen deficient rats and the possible mechanism, laying the groundwork for further use in patients with postmenoposal osteoporosis.Methods41six-months female SD rats were randomly divided into five groups, including normal control group(n=9), model blank group (n=8), Qiang Gu Kang Shu(QGKS) low-dose group(n=8), QGKS high-dose group(n=8), estrogen group(n=8). Preparation of ovariectomized rats in model blank group, QGKS low-dose group, QGKS high-dose group, estrogen group. After ovariectomizing, respectively, to feed low-dose or high dose QGKS to two QGKS groups, and to feed estrogen to estrogen-control group. At the same time to feed saline to normal control group and model blank groups for one month, respectively. Then, the rat BMD of bilateral femora in each group is measured by GE lunar Bone Densitometer, and micro structure of femoral trabecular bone is observed with light and electron microscope by using an image analysis system for bone histomorphometry parameters. The protein expression of OPG and RANKL in bone marrow stroma cells is detected by immunohistochemistry. The experimental data is expressed by mean+standard deviation (x±s), and the comparison between QGKS groups and controls was performed by single factor analysis of variance (ANOVA) in SPSS statistical software.Results (1) Measurement of BMD:compared with normal control rats, the’femoral BMD in the model blank group was significantly decreased (P<0.01), indicating that the osteoprosis model is successfully prepared by ovariotomy. Compared with the model blank group, BMD in estrogen group and QGKS groups (low and high) was significantly increased (P<0.05). Compared with estrogen prevention group, BMD in both low-and high-dose groups was slightly less, however the difference was not significant (P>0.05). The difference of BMD between QGKS low-and high-dose groups was also not significant (P>0.05).(2) Bone trabecula morphological changes under light and electron microscopy:in model blank group was observed the reduced and attenuated bone trabecula, accompanied with enlarged bone marrow cavity, rupture and discontinue of bone trabecula. Compared with the model group, in estrogen controls and in QGKS low-and high-dose groups the bone trabecula arranged normally with increased width of trabecular bone and reduced spacing.(3)Bone histomorphometry parameters:compared with normal control group, the bone trabecula thickness, area and area percentage of model blank group were significantly decreased (P<0.01), the spacing of bone trabecula was significantly increased (P<0.01). Compared with the model blank group, the trabecular bone thickness, area and area percentage in estrogen controls and both QGKS low-and high-dose groups were significsntly increased accompanied with reduced trabecular bone spacing (P<0.01, respectively). In both QGKS low-and high-dose groups the difference was not statistically significant (P>0.05).(4)Protein expression of RANKL/OPG:over-expression of RANKL in bone marrow stromal cells was observed in model blank group by immunohistochemistry. Compared with the model blank group, the level of RANKL in the bone stromal cells from estrogen and QGKS feed rats was much lower. Inversely, expression of OPG in bone marrow stromal cells from model blank rats reduced dramatically. Compared with the model blank group, OPG expression in bone marrow stromal cells from both estrogen-feed reats and QGKS-feed rats increased significantly(P<0.05, respectively).Conclusion In ovariectomized rats the femoral BMD and the volume of bone trabecula decreased accompanied with damaged micro-structure, which can mimic human postmenopausal osteoporosis. Qiang Gu Kang Shu capsules can prevent osteoporosis from ovariotomy in rats by maintaining bone mass and microstructure of trabecular bone. The effect of this formula on bone metabolism mainly based on the inhibiton of RANKL expression and induction of OPG production in bone cells. These results suggest that traditional Chinese medicine Qiang Gu Kang Shu capsules, prepared by Bu Shen Zhuang Gu method, might be a potential formula in treatment of postmenopausal osteoporosis. |
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