The Effects Of The Methods Of Nourishing The Kidney-yin And Warming And Recuperating The Kidney-yang On The Expression Of OPG, RANKL In Osteoblasts And Marrow Matrix Cells Of The Model Rats Established By Bilateral Ovariectomy

The Effects of the methods of nourishing the Kidney-yin and warming and recuperating the Kidney-yang on the Expression of OPG, RANKL in Osteoblasts and marrow matrix cells of the Model Rats established by bilateral ovariectomyObjectiveTo observe the effects of the methods of nourishing the kidney-yin and warming the kidney-yang on the expression of OPG, RANKL in osteoblasts and marrow stroma cells of the model rats, which are established by bilateral ovariectomy so as to explore the mechanism of preventing osteoporosis by nourishing the kidney-yin and warming the kidney-yang.Material and MethodsFirst, sixty-six female Wistar rats were selected and divided into six groups randomly: ten in the normal control group, ten in the pseudo-operation group and forty-six 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.Then, 2 weeks after operation, the rats of the operation group were divided into four sub-groups again: eleven in model group, eleven in stilbestrol group, twelve in nourishing the Kidney-yin group and twelve in warming and recuperating the Kidney-yang group. Stilbestrol, nourishing the Kidney-yin group and warming and recuperating the Kidney-yang group were administrated orally. Distilled water was administrated orally to normal control group, pseudo-operation group and model group at the same time. The drugs or water were given once a day and last six days. There has 1 day pause every six days. The whole period last 3 months. At the third day and the sixteenth 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.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 paraffin tissue section after being decalcified to observe the protein and mRNA expression of OPG, RANKL by means of immunohistochemistry and in situ hybridization.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 Zuogui pill and Yougui pill. Zuogui pill and Yougui pill can both reduce the bone's high transformation.2．Compared with nomal control group, the protein and mRNA expression of RANKL significantly increased ,while the protein and mRNA expression of OPG decreased after the bilateral ovaries of the rats were extirpated. After 3 months, compared with model group,OPG significantly increased while RANKL significantly decreased in warming and recuperating the Kidney-yang group; OPG significantly increased while no significant change of RANKL expression in nourishing the Kidney-yin group. Conclusions1．The methods of nourishing the kidney-yin and warming and recuperating the kidney-yang can prevent and treat osteoporosis caused by bilateral ovariectomy, but the effect of warming and recuperating the kidney-yang is better than that of nourishing the kidney-yin.2．Warming and recuperating the kidney-yang can prevent and treat osteoporosis by enhancing the expression of OPG in OB and bone matrix while inhibiting the expression of RANKL in OB and bone matrix significantly. The method of nourishing the kidney-yin can only enhance the expression of OPG , then the combination of OPG and RANKL increases so as to inhibite the activity of OC. One of the mechanism is that the former can regulate both OPG and RANKL, while the latter can only regulate the OPG...