The Regulation Of Part A Extracted From Zhuangguzhitong Decoction In Alcohol On Estrogen Relevant Factors In Alcohol On Estrogen Relevant Factors In Ovariectomized Female Rats With Osteoporosis

Objective1. To discuss the relevance of osteoporosis to estrogen and its correlation factors.2. To discuss the pharmacodynamic mechanism of part A extracted from Zhuangguzhitong Decoction in alcohol on osteoporosis, mainly observe the Chinese herbs'effects of hormone levels, serum biochemical parameters, bone mineral density and bone biomechanics, bone formation,bone resorption and its related cytokine proteins and gene expression of ovariectomized female rats.MethodsThe first part Literature ReviewAdopting computer network technology, as well as literature search and review of epidemiological research methods, to study and analyse the etiology and pathogenesis, clinical classification, treatment, therapies and current research progress of troditional Chinese medicine in preventing and treating osteoporosis. Numerous researches show that treating osteoporosis with traditional Chinese herbs is effective with little side effects and no drug resistance, it is an area worthy of exploration.The second part Experimental research of part A extracted from Zhuangguzhitong Decoction in alcohol on ovariectomized female rats with osteoporosis.Chose 75 three months' SD female rats weighing 250g,which were randomly divided into 5 groups:control group(n=15), sham operation group (n=15), model group(n=15), Zhuangguzhitong Decoction group(n=15), part A extracted from anhydrous alcohol group A(n=15). When taking out stitches 5 days after operation, starting administering medications for 13 weeks. Studied its efficacy with blood and bone samples from the bone mineral density,bone biomechanics,pathology, immunohistochemical method (SABC) and situ hybridization were used to detect estrogen-related cytokines protein and gene expression of cytokines and so on. Adopt radioimmunoassay and enzyme-linked immunosorbent assay from serum E2, PTH, CT, BGP, ALP and so to study its mechanism of action.ResultsThe first part Literature ReviewBone is the location of osteoporosis, and has closely relationship with the liver and kidney. The deficiency of kidney qi and lack of yin fluid of the liver and kidney are essential of the disease, and bone stasis are indications. The treatment shoud comply with "tonifying the liver and kidney can make muscles and bones strong "theory in the "Ben Cao Jing Shu ". and adopt tonifying the kidney to make bone strong, filling lean marrow, tonifying the liver to strong muscle,promoting blood circulation to stop pain.According to Chinese medicine, "Kidneys advocate bone"theory, as tested factors, zhuangguzhitong Decoction is curative in clinic.The second part Experimental research of part A extracted from Zhuangguzhitong Decoction in alcohol on ovariectomized female rats with osteoporosis.1. The influence of part A extracted from Zhuangguzhitong Decoction in alcohol on the serum biochemical parameters of ovariectomized female rats.Compared with model group, Zhuangguzhitong Decoction can significantly elevate the levels of serum E2, CT, BGP, lower serum PTH, ALP levels (p<0.05); serum CT was significantly higher in Alcohol A group (p<0.05), serum ALP decreased significantly (p <0.05); Alcohol A group can increase serum E2, BGP but has no statistical significance (p>0.05); Alcohol A group can lower serum PTH but has no statistical Significance (p> 0.05).2. The influence of part A extracted from Zhuangguzhitong Decoction in alcohol on the bone mineral density and bone biomechanics of ovariectomized female rats.Compared with model group, bone mineral density and lumbar spine bone mineral density increased significantly in the whole decoction group and Alcohol A group,and the difference has statistically significance (p<0.05). Compared with model group, the whole decoction group and Alcohol A group can increase the maximum bending strength and tibia maximum stress significantly, the difference has statistically significance (p<0.05).3. The influence of part A extracted from Zhuangguzhitong Decoction in alcohol on the bone tissue and ultrastructure of pathologic of ovariectomized female rats. Routine pathological sections of bone tissue were observed under light microscope and the results were as follows.Compared with model group,the whole decoction Group and and Alcohol A group'distribution of bone trabecular density is equality, trabecular and marrow ratio returned to normal, osteoblasts can be observed under microscopeTEM ultrastructure of bone tissue results were as followsCompared with model group.the whole decoction Group and and Alcohol A group has normal mitochondria in bone cells, no significant changes in bone cell apophysis, bone lacuna reduced so significantly that appears flat oval.4. The influence of part A extracted from Zhuangguzhitong Decoction in alcohol on estrogen relevant factor-- protein of ovariectomized female rats.Using Immunohistochemical method (SABC) to detecte the expression of MMP-2, OPG, in model group,compared with the control group, the levels were significantly lower, the difference has statistically significance(p<0.05); Compared with the model group, the MMP-2, OPG protein levels in the whole decoction Group and Alcohol A group were significantly higher, the difference has statistically significance(p<0.05). Compared with the control group,the protein expression levels of MMP-9, IL-6 in model group were significantly higher, the difference has statistically significance(p<0.05); Compared with the model group, the levels in the whole decoction Group and Alcohol A group were significantly lower, the difference has statistically significance(p<0.05).5. The influence of part A extracted from Zhuangguzhitong Decoction in alcohol on estrogen relevant factor—gene of ovariectomized female rats.Testing TGF-β1mRNA, with the method of Situ hybridization, compared with the control group, in model group, I collagen mRNA expression was significantly reduced, the difference has statistically significance(p<0.05);compared with the model group expression was Increased in the whole decoction Group and Alcohol A group,and the difference has statistically significance (p<0.05).Conclution1. In traditional Chinese medicine, the deficiency of kidney qi and lack of yin fluid of the liver and kidney were essential of the disease, and bone stasis were indications. According to the theory that "Kidneys advocate bone"theory. Studying Zhuangguzhitong Decoction's effect on preventing and treating osteoporosis, regulating the balance of estrogen-related factors of ovariectomized female rats, promoting bone formation and inhibiting bone resorption to make the bone reconstruction be carried out.2. Of ovariectomized female rats, the serum E2, CT, BGP, ALP were decreased, the serum PTH was increased, the femoral and lumbar bone density were decreased, both maximum stress and bending strength of the tibia were declined.In bone tissue,the expression of bone formation factors MMP-2, OPG were reduced but bone resorption associated factor MMP-9, IL-6 were increased, TGF-β1mRNA and collagen type I mRNA' expression were decreased. So as to promote bone resoiption and inhibit bone formation, cause osteoporosis. 3. Part A extracted from Zhuangguzhitong Decoction in alcohol can increase serum E2, CT, BGP, ALP, decrease serum PTH levels, it can increase the femur and lumbar spine bone mineral density, increase the maximum bending strength and tibia maximum stress,increase the expression of bone formation factors MMP-2, OPG, Part A extracted from Zhuangguzhitong Decoction in alcohol can decrease the bone resorption associated factor MMP-9, IL-6, increase TGF-β1mRNA and collagen typeⅠmRNA' expression. It reduced bone turnover.promoted bone formation and inhibited bone resorption, improved the molecular structure of bone matrix and strengthened the biomechanical properties of bone in treating osteoporosis.