| ObjectiveTo explore the effects of Herba taxilili, Herba ecliptae, Fructus schisandrae chinensis and Rhizomaof ,which has the role of nourishing the kidney-yin, on osteoporosis model rats established by bilateral ovariectomy by means of bone histomorphometry. To reveal the characteristics of effective mechanism of the four Chinese medicine, and to provide further experimental evidence for clarifying the mechanism of effects of Chinese medicine on osteoporosis and the traditional Chinese medicine theory of " The kidney dominating bone " by focusing on the osteoclast differentiation and regulatory factors and hormones and cytokines related to bone metabolism closely.Material and MethodsFirst, ninty four female Wistar rats were selected and divided into three groups randomly: twelve in the normal control group, twelve in the pseudo-operation group, seventy in the ovariectomy (OVX) group. After anesthesia, both ovaries of all the rats in the operation group were extirpated, while only the fatty tissue nearby the ovaries of the rats in the pseudo-operation group were extirpated. 1 month after operation, the rats of the operation group were divided into six sub-groups again: twelve in model group, twelve in positive control group, twelve in Herba ecliptae group, twelve in Rhizoma dioscoreae, eleven in Herba taxilili and eleven in Fructus schisandrae chinensis.Rats in Herba taxilili group, Herba ecliptae group, Fructus schisandrae chinensis group and Rhizomaof group were administrated orally the decoction of corresponding Chinese medicine (decoction concentration was 1g/ml, the delivery volume was 5.6ml/kg weight and the dose was 5.6g/kg weight). Rats in positive control group were administrated orally Stilbestrol ,the drug concentration was 0.008mg/ml, the delivery volume was 5.6ml/kg weight and the dose was 0.0448mg/kg weight. The drugs were given once a day and last six days. There has 1 day pause every six days. The whole period last 3 months. While rats in normal control group, pseudo-operation group and model group were administrated orally the equal volume of distilled water. At the fifteenth day and the third day before the day all the rats were killed, hydrochloric acheomycin were respectively injected into the abdomen of the rats according to 30mg/kg to mark the bone with fluorescence. Then all them were sacrificed to take blood for testing the serum contents of IL-1, IL-6, BGP, CT, and PTH in each specimen by means of enzyme linked immunosorbent. After being killed, the right tibia of the rats were retained for bone tissue section which weren't decalcified to evaluate the change of bone histomorphometry; at the same time the left tibia of the rats were used for frozen tissue section after being decalcified to observe the protein expression of OPG, RANKL by means of immunohistochemistry.Statistical analysis : results are presented as Mean±SD. Differences between groups were analyzed using q test by one-way ANOVA.Results1. After the bilateral ovaries of the rats were extirpated, TBV% of tibia significantly decreased, while TRS% and TFS% significantly increased, which indicated that the model established by extirpating bilaterral ovaries is a kind of high transformed osteoporosis model which has higher bone resorption than bone formation. Compared with model group, TBV% significantly increased while TRS% significantly decreased, after being treated by prescriptions of Herba taxilili, Herba ecliptae, Fructus schisandrae chinensis and Rhizoma dioscoreae, which all could reduce the bone's high transformation.2. Compared with nomal control group, the protein expression of RANKL significantly increased , while the expression of OPG decreased after the bilateral ovaries of the rats being extirpated. After 3 months, compared with model group, the expression of OPG increased significantly while the expression of RANKL decreased significantly in Fructus schisandrae chinensis group and Rhizoma dioscoreae group; in Herba taxilili group, the expression of OPG increased significantly while there was no significant change in expression of RANKL; in Herba ecliptae group, the expression of RANKL significantly decreased while there was no significant change in the expression of OPG.3. Compared with nomal control group, the serum content of IL-1 and IL-6 significantly increased after the bilateral ovaries of the rats being extirpated. After 3 months, compared with model group, the level of IL-1 and IL-6 in peripheral blood serum significantly decreased in Fructus schisandrae chinensis group and Rhizoma dioscoreae group; the serum level of IL-6 significantly decreased while there was no significant change in the serum level of IL-1 in Herba ecliptae group; and in Herba taxilili group, the level of IL-1 significantly decreased while the level of IL-6 in peripheral blood serum was no significant change.4. Compared with nomal control group, the serum content of BGP significantly increased, while CT significantly decreased after the bilateral ovaries of the rats being extirpated. After 3 months, in Fructus schisandrae chinensis group and Rhizoma dioscoreae group, compared with model group, the serum content of BGP decreased while CT significantly increased; the serum content of CT significantly increased while there was no significant change in BGP content in Herba taxililiand group and Herba ecliptae group. Moreover, there was no significant change in the serum content of PTH in all groups.Conclusions1.Rat Ovary removal could cause of a high-transformational model of osteoporosis, bone resorption was greater than the formation. Herba taxilili, Herba ecliptae, Fructus schisandrae chinensis and Rhizomaof could inhibit osteoporosis to some extent, and let it regain the relative balance of the bone formation greater than bone resorption, so as to achieve the role of treatment of osteoporosis.2.Herba taxilili, Herba ecliptae, Fructus schisandrae chinensis and Rhizomaof could achieve the role of treatment of osteoporosis by regulating certain aspects the osteoclast differentiation and regulation factors and the cytokines and hormones which closely related to bone metabolism. And they had shown different characteristics in the role of the treatment, as follows:Fructus schisandrae chinensis and Rhizomaof could enhance the expression of OPG and inhibit the expression of RANKL in osteoblast and bone marrow stromal cells; they also could reduce the level of IL-1, IL-6 and increase the level of CT in the peripheral blood. Herba taxilili could enhance the expression of OPG, reduce the level of IL-1 and increase the level of CT in the peripheral blood, but it had no effect on the expression of RANKL and the level of IL-6. Herba ecliptae could reduce the expression of RANKL ,decrease the level of IL-6 and increase the level of CT in the peripheral blood, but it had no effect on OPG and IL-1. The above Chinese medicines had no significant effect on PTH in the peripheral blood. |
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