Experimental Research On Signal Transduction Mechanism Change Of BMP-4 Of Osteoporosis Due To Kidney-deficiency

Purpose: To study the relativity of pathogenesis of osteoporosis due to kidney deficiency and disorganization of signal transduction mechanism of bone formation induced by BMP-4. To explore the adjustable function of Nanometer calcium compound Chinese medicine to the disorganization of signal transduction mechanism, which is tonifying kidney-Qi and strengthening essence, supplying calcium to bone and invigorating the kidney to make the bone strong.Materials and Method: The experiment includes two parts: in vivo part and in vitro part. In vivo part, we established the rat model of osteoporosis due to kidney-deficiency, used the method of ovarie-ctomy, had treated the osteoporosis rat with experimental drugs (big dose, middle dose, small dose group) for 12 weeks,which is tonifying kidney-Qi and strengthening essence, invigoratingthe kidney to make the bone strong. Simultaneously,used Gush-ukang granule and Oyster shell carbonate chewable tablets as positive control groups. And rats in the normal group and model group were not given any treatment. We had used dual ener-gy X-ray bone density instrument to detect the bone mineral density of the rats; detected the Alkaline phosphatase, tartrateresistant acid phosphatase in the blood serum with the method efbiochemistry.We had used the RT-PCR, western blotting method to investigate the expression of BMP-4, Smad5, Smad6 mRNA and proteins. In vitro part, we cultured the osteoblastof the newborn rats calvarias: identified the osteoblast cultured in v-itro with the method of enzyme cytochemistry(ALP- Gomori),by the method of serum pharmacol- ogy, cultured osteoblast in rats serum which preventing and treating 8 weeks by Nanometer calcium and tonic-kidney compound Chinese drugs, then evaluated the proliferation of the osteoblast by MTT method; detected the expression of mRNA of BMP-4, Smad5,Smad6 gene were detected by RT-PCR.Result: 1. In vivo part: (1) The comparison of the BMD of the femur of the experimental rats: comparing with the normal group, the BMD of the femur has evident tendency to reduce; comparing with the model group, in the experimental drug groups(big dose> middle dose> small dose group) and the Gushukang granule group, the Oyster shell carbonate chewable tablets group, the BMD of femur increased evidently. (2) The comparison of the blood biochemistry index of the experimental rats: Comparing with the normal group, the activity of TRAP and ALP in the blood serum of model group increased evidently; Comparing with the model group, the activity of TRAP and ALP in the blood serum of the experimental drug groups(big dose^ middle dose^ small dose group), Gushukang granule group and Oyster shell carbonate chewable tablets group decreased in some extent, but the experimental drug group with big dose and Gushukang granule group, comparing with model group, have no significance in statistics. While the other groups, comparing with model group, not only have significance in statistics, but also reach the level of normal group. (3) The comparison of uterus exponent of the experimental animals: the uterus exponent of the model group is lower than that of the normal group evidently, the experimental drug groups(big dose, middle dose^ small dose group). Oyster shell carbonate chewable tablets group and Gushukang granule group can increase the uterusexponent only in some extent but have no significance in statistics. (4) RT-PCR and Western blotting detection: theBMP-4 and its signal transduction factor Smad5, Smad6 exist in the normal bone tissue. Comparing with the normal group, the level of the expression of the gene and proteins of BMP-4 and Smad5 in the bone tissue of model group decreased. After the precaution treatment with the experimental drug groups(big dose> middle dose> small dose group), Gushukang granule and Oyster shell carbonate chewable tablets group can increase the mRNA and protein expression level significantly, the level of the expression of the gene and protein of smad6 of model group increased, the treatment of the experimental drug gr...