| Purpose: To study the dependablity of the pathogenesis of osteoporosis due to kidney deficiency and the influence of Smurf1 and BMP7/Smads protein in the kidney,femur,hypothalamus. To explore the adjustable function of Nanometer calcium compound Chinese medicine to the mechanism, which is tonifying the kidney to consolidate the essence and strengthening bone and marrow, supplying calcium to bone and invigorating the kidney to make the bone strong.Material and method: Using the medthod of ovriectomy,we established the rat model of osteoporosis due to Kidney-deficiency; treated the osteoporosis rat with experimental drugs (high dose, small dose group) for 12 weeks, which is tonifying the kidney to consolidate the essence and strengthening bone and marrow , invigorating the kidney to make the bone strong. Simultaneously, used Gushukang granule and Gaitianli tablets and Buzhongyiqi herb as positive control groups. And rats in the normal group and model group were not given any treatment. We had used dual energy X-ray bone density instrument to detect the bone mineral density of the rats;observed configuration of the kidney, hypothalamus,femur tissue with the method of HE stain, We had used the IHC method to investigate the expression of smad1/5,BMP7,Smurf1 proteins in the kidney, hypothalamus,femur, hypothalamus of the rats.Results1.BMD data of the left femur of rats: model gourp got an obviously decrease of the level of BMD (p<0.01);and all the medicines groups (nanometer calcium kidney-tonifying herbs, Gushukang granule and Gaitianli and Buzhongyiqi herb) have increased the level of BMD compare with the model groups, while the high dose groups has got the best effects(p<0.01).2. The comparison of uterus exponent of the experimental rats: the uterus exponent of the model group is lower than that of the normal group evidently, all the medicines groups can increase the uterus exponent only in some extent but have no significance in statistics.3. The comparison of the morphometry :of the femur and uterus and vagina tissues in the experimental rats:We observed the HE stain femur parts of the experimental the rats:in normal group, the shape and structure of the bone trabecula are complete, and the arrangement are.normal;In the model group, the arrangement of bone trabecula structure is rarefaction, the bone trabecula thickness. decreased, the marrow cavity area increased obviously;while in all the medicines groups . We observed the HE stain uterus parts of the experimental the rats:In normal group, configuration of the cornua uteri were normal, In the model group, configuration of the cornua uteri were shrinking, all the medicines groups can reform configuration of the cornua uteri compare with the model groups.We observed the HE stain vagina parts of the experimental the rats: In normal group, epithelium of the vagina were thick and normal;In the model group, epithelium of the vagina were thin .4.Immunohistochemical method to detect the BMP7The BMP7 protein expression in the femur of rats : model groups got a decreasd in BMP7 protein expression compared with normal groups ,but no meaning in the statisical comparison. all medicine groups increasd BMP7 protein expression to differrent level compared with model groups, but no meaning in the statisical comparison.The BMP7 protein expression in the kidney of rats : model groups got a obviously decreasd in BMP7 protein expression compared with normal groups (p<0.01). all medicine groups increasd BMP7 protein expression to differrent level compared with model groups, nanometer calcium kidney-tonifying herbs gruops with high dose get the highest increase(p<0.01).The BMP7 protein expression in the hypothalamus of rats : model groups got a obviously increasd in BMP7 protein expression compared with normal groups (p<0.01). all medicine groups decreasd BMP7 protein expression to differrent level compared with model groups,(p<0.01).5. Immunohistochemical method to detect the Smad1/5 :The Smad1/5 protein expression in the femur of rats : model groups got a obviously decreasd in Smad1/5 protein expression compared with normal groups (p<0.01). all medicine groups increasd Smad1/5 protein expression to differrent level compared with model groups, nanometer calcium kidney-tonifying herbs gruops with high dose get the highest increase(p<0.01).The Smad1/5 protein expression in the kidney of rats : model groups got a obviously decreasd in Smad1/5 protein expression compared with normal groups (p<0.01). all medicine groups increasd Smad1/5 protein expression to differrent level compared with model groups, nanometer calcium kidney-tonifying herbs gruops with high dose get the highest increase(p<0.01).The Smad1/5 protein expression in the hypothalamus of rats : model groups got a obviously increasd in Smad1/5 protein expression compared with normal groups (p<0.01). all medicine groups decreasd Smad1/5 protein expression to differrent level compared with model groups, nanometer calcium kidney-tonifying herbs gruops with high dose get the highest decrease(p<0.01).6.Immunohistochemical method to detect the Smurf1:The Smurf1 protein expression in the femur of rats : model groups got a decreasd in Smurf1 protein expression compared with normal groups (p<0.01). all medicine groups increasd Smurf1 protein expression to differrent level compared with model groups, nanometer calcium kidney-tonifying herbs gruops with high dose get the highest increase(p<0.01).The Smurf1 protein expression in the femur of rats : model groups got a obviously decreasd in Smurf1 protein expression compared with normal groups (p<0.01). all medicine groups increasd Smurf1 protein expression to differrent level compared with model groups, nanometer calcium kidney-tonifying herbs gruops with high dose get the highest increase(p<0.01).The Smurf1 protein expression in the femur of rats : model groups got a obviously increasd in Smurf1 protein expression compared with normal groups (p<0.01). all medicine groups decreasd Smurf1 protein expression to differrent level compared with model groups,(p<0.01).Conclusion:1.The rat model of osteoporosis due to kidney-deficiency can be established successfully by the method of ovariectomy.2.Nanometer calcium kidney-tonifying herbs can distinctly increase the bone density3.In normal rats, kidney,the epiphysis of femur, hypothalamus can express the BMP7 ,and its signal transduction molecule Smad1/5 in the protein level.That means kidney , femur ,hypothalamus tissues in normal rats exist the pathway of Smad1/5 of BMP7.4.The decreasing of Smad1/5 and BMP7 and Smurf1 protein expression in the kidney and femur of rats while the increasing of Smad1/5 and BMP7 and Smurf1 protein expression in hypothalamus ,may be the one of the mechanism of osteoporosis due to kidney deficiency.5.The level of Smad1/5 and BMP7 and Smurf1 protein expression in the kidney and femur of rats can obviously increased by nanometer calcium kidney-tonifying herbs, while the level of Smad1/5 and BMP7 and Smurf1 protein expression in hypothalamus had obviously decreased by nanometer calcium kidney-tonifying herbs ,which indicated the chinese herbs with kidney-tonifying,calcium-supplying action are able to prevent and cure osteoporosis due to kidney deficiency indirectly by inducing the expression of Smad1/5 and BMP7 and Smurf1 in the kidney and femur,and restraining the expression of Smad1/5 and BMP7 and Smurf1 in the hypothalamus.
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