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Effect of Huoxuebushen Decoction on Expression of VEGF and OPG/RANKL in Steroid-induced Osteonecrosis of the Femoral Head

Author: YanBing
Tutor: WangHeMing
School: Fujian University of Traditional Chinese Medicine
Course: TCM orthopedics and traumatology
Keywords: Femoral head necrosis Huoxue Bushen soup Vascular endothelial growth factor OPG Receptor activator of nuclear factor -kB ligand
CLC: R285.5
Type: PhD thesis
Year: 2010
Downloads: 202
Quote: 0
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Abstract


Objective: In this study, through animal experiments, tissue morphology and cellular and molecular level, the study Huoxue Bushen decoction on the expression of VEGF and OPG, RANKL investigate the pathogenesis of femoral head necrosis and Huoxue Bushen Decoction hormone The mechanism of action of the femoral head necrosis, experimental and theoretical basis for promoting blood kidney soup therapeutic principle. Research methods. Huoxue Bushen decoction on rabbit femoral head necrosis of VEGF 1.1 130 New Zealand white rabbits were divided into five groups: normal control group, the hormone model group, high (12.88g/kg · d), medium (9.66 g / kg · d), low (6.44g/kg · d) dose Huoxue Bushen decoction treatment group (modeling Matsui modeling method). Normal control group and hormone model group, saline daily l0ml gavage. Huoxue Bushen decoction of Chinese medicine treatment group given different concentrations gavage. After continuous administration is based in batches at 8 and 12 weeks. 1.2 rabbit femoral head morphology observed under the microscope and empty lacunae count. 1.3 Determination of vascular endothelial growth factor (VEGF) by immunohistochemistry SP method. 2. Huoxue Bushen soup rabbit femoral head necrosis impact of serum OPG / RANKL 2.1 Animal grouping, modeling the same administration. Each group were sacrificed at 4, 8 and 12 weeks of 2.2 administered eight white rabbits abdominal aortic blood 3ml serum collected after centrifugation, save spare. 2.3 using a double antibody sandwich ABC-ELISA method detects rabbit serum of OPG, of RANKL content. Huoxue Bushen Decoction Serum the rat osteoblasts the OPG / RANKLmRNA expression neonatal SD rats calvaria 3.1 to take into bone cells in primary and subcultured. The take 3 substituting Growing into bone cells, osteoblasts were identified microscope microscope. 3.2 SD rats 12, divided into four groups (saline control group, high, medium and low dose Huoxue Bushen decoction of Chinese medicine group) (n = 3). 3 ml saline control rats, high in the low dose group rats at different concentrations of of 3ml blood circulation and kidney soup gavage, twice a day for three consecutive days. Femoral artery blood two hours after each group in the last administration. After centrifugation, serum was collected, stored for use. The rat osteoblast 3.3 were placed in culture medium containing serum (20% concentration), placed in the incubator. 3.4 RT-PCR method of osteoblasts, OPG RANKLmRNA expression semi-quantitative detection. Observed results 1.HE hormone model group: trabecular sparse, structural disorder, a significant reduction of bone marrow cells, bone marrow and more filled with fat cells, bone cells reduce the number of empty lacunae increased; the medicine treatment groups in the bone of the experimental period The beam structure is basically complete. High, 8 and 12 weeks in the dose of Chinese medicine group trabecular surface formation of new bone cells and bone tissue. 2.VEGF expression of four weeks, high, medium and low dose group and model group VEGF expression was significantly higher than the control group (P lt; 0.01). In the low-dose group VEGF expression than the model group (P lt; 0.05); 8 weeks, high in the low dose group VEGF expression was significantly higher than the model group (P lt; 0.01); 12 weeks high, low dose of VEGF expression was significantly higher than that in the model group (P lt; 0.01). 4 weeks 3.OPG, RANKL serum levels: high, low dose group, normal control group OPG levels were significantly higher than the model group (P lt; 0.01). Week 8: model group OPG content was significantly lower than the high in the low dose group and normal control group (P lt; 0.01). High, medium and low dose group was significantly higher than the normal control group (P lt; 0.01). Week 12: high, low dose group, normal control group OPG levels were significantly higher than the model group (P lt; 0.01). RANKL content of 8 and 12 weeks of the model group were significantly higher than the high, the low-dose group and normal control group (P lt; 0.01). 4 osteoblast cells the OPG, RANKLmRNA expression values, the medium dose traditional Chinese medicine group OPGmRNA expression value higher than the saline control group (P lt; 0.05). Was no significant difference between the high, medium and low dose group. RANKLmRNA expression values: high, medium dose traditional Chinese medicine group was significantly lower than the saline control group (P lt; 0.01). The low dose group than in the saline control group (P lt; 0.05). High dose group than in the low dose group (P lt; 0.05). Conclusion 1. Femoral head necrosis local expression of VEGF decreased inhibition of the blood vessels and bone regeneration. 2. Huoxue Bushen soup enhanced local expression of VEGF necrosis of the femoral head. 3. Decline in femoral head necrosis rabbit serum OPG content of RANKL levels rise, OPG / RANKL ratio decreased. Huoxue Bushen soup to improve the content of serum OPG, reducing RANKL content, increased OPG / RANKL ratio. Huoxue Bushen soup upregulated the expression of OPG mRNA down RANKL mRNA expression, OPG / RANKL drug targets for blood circulation and kidney soup.

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