| 1. ObjectiveIn 3 months, with the experiment and systematic observation to study the effect of single Chinese herb Rhizoma Drynariae,Psoralea Corylifolia and the bi-combined on bone metabolism of lumbar and femur in ovariectomized ratmodel of osteoporosis.And we take alendronate as a positive control to compare with these two kinds of Chinese traditional medicine on the effect characteristics of bone metabolism.2. MethodWe select 72 four-month-aged unmated female SD rats, weight within 250± 25 g.60 rats were randomly selected from the mold to take the surgury of bilateral ovary removal and the remaining were into false operation group. Then the model group was randomly evenly divided into 5 groups : model group, alendronate group,Psoralea Corylifolia group, Rhizoma Drynariae group and bi-combined group. They were given intragastric administration after normal feeding for 4weeks. According to body surface area conversion in human, the alendronate group were given alendronate sodium 6.87mg/kg, Similarly, 1g/kg was given in Psoralea Corylifolia group,Rhizoma Drynaria group and bi-combined group. And according to the weight,the false operation group and model group were given the same volume of distilled water daily for 12 weeks. After the experiment,we took the Arterial blood to test ALP,IL-1and IL-6.And the proximal 1/3 of right femur and lumbar spine(L5) was made to decalcified bone tissue sections to compare the trabecular bone in each group observed by microscope.Then we use the Image-pro plus 5 software for bone histomorphometry data acquisition and analysis. After using dual energy X-ray bone density instrument for bone density scans in the left femur and lumbar spine(L4),we send the left femur for biomechanical test and use three point bending test method to test and analysis its biomechanical properties.3. Result1.The general living performance of rats: the model group rats is dispirited in spirit,less shiny in hair and poor in vitality. The single Chinese herb and bi-combined group compared with model group, the ability of activity is stronger,the reaction is more sensitive and the hair is more lustrous. They also act more docile and stable in emotion in the intragastric administration,2. The weight change: Compared with the false operation group, the weight of model group gain 20% in 4 weeks after modeling. The weight of the Rhizoma Drynariae group, Psoralea Corylifolia group and bi-combined group increase by weeks of age.3. Microscopic observation on bone tissue biopsy specimens of 3 groups:Compared with the false operation group, the trabecular bone of model group decreased markedly, arranged more sparse and irregular, appeared fracture partly.Rhizoma Drynariae, Psoralea Corylifolia, bi-combined group and western medicine group compared with the model group, we find that the bone trabecula distribute more evenly, thicker, smaller in spacing,and with good reticular structure.4.Analysis of 4 groups of indexes of bone histomorphometry:4.1 Compared with false operation group, the percentage of trabecular bone area(Tb.Ar%) of model group decreased by 53.3%.4.2 Comparing with the model group, the femur and lumbar vertebrae Tb.Ar% of western medicine group, Rhizoma Drynariae group, Psoralea Corylifolia group,bi-combine d group respectively increase 98.7%, 49.8%, 47.6%, 61.6%. 4.3 Compared with false operation group, the trabecular bone volume(Tb.N) in model group decreased by 55.4%.4.4 Compared with the model group, the Tb.N of lumbar vertebrae and femur in western medicine group, Rhizoma Drynariae group, Psoralea Corylifolia group,bi-combined group increased respectively by 91.1%, 44.9%, 41.3%, 62.5%.4.5 Compared with false operation group, the trabecular separation(Tb.Sp) of model group, increased by 55.7%.4.6 Compared with the model group, the femur and lumbar vertebrae in Tb.Sp in western medicine group, Rhizoma Drynariae group, Psoralea Corylifolia group,bi-combined group decreased respectively :39.6%, 26.1%, 25.7%, 30.5%.5 The BMD and BMC values of femur and lumbar vertebrae in different groups of rats:5.1 Compared with the false operation group, BMD of model group decreased by36.3%.5.2 Compared with the model group,the BMD of femur and lumbar vertebrae in western medicine group, Rhizoma Drynariae group, Psoralea Corylifolia group,bi-combined group rat increased respectively by 60.1%, 34.2%, 35.6%, 44.1%.5.3 BMC in model group reduce by31.3% compared with the false operation.Likely,the BMC of femur and lumbar vertebrae in western medicine group, Rhizoma Drynariae group, Psoralea Corylifolia group, bi-combined group were increased by45.4%, 26.2%, 29.1%, 35.4%,comparing with the model group.6.Detection of femur biomechanic indicators in 6 groups: compared with the false operation group,the Young’s modulus maximum load, stiffness in model group decreased respectively by 28.3%, 25.2%, 43%.And the large load, the young’s modulus of rat femur, stiffness in western medicine group, Rhizoma Drynariae group, Psoralea Corylifolia group and bi-combined group compared with the model group, increased significantly.7.The content of peripheral ALP, IL-1, IL-6 in serum of 7 groups: compared with the false operation group, the content of ALP in peripheral serum increased 77.5% in model group.Compared with the model group, ALP content in peripheral serum of the western medicine group, Rhizoma Drynariae group, Psoralea Corylifolia group and bi-combined group decreased by 36.7%, 28.4%, 32.2%, 29.1%. Compared with the false operation group, IL-1, IL-6 in the peripheral serum increased by 31.3%, 55.7% in model group. The IL-6 content in serum of western medicine group, Rhizoma Drynariae group, Psoralea Corylifolia group and bi-combined group decreased by37.8%, 25.1%, 25.8%, 34.1% and the IL-1 content in serum of western medicine group,Psoralea Corylifolia group, bi-combined group decreased by 38.9%, 33.2%, 32.2%compared with the model group. However, Rhizoma Drynariae group had no significant change;.4. Conclusion1.through bilateral ovary removal surgury on rats,we can make ideal animal model of osteoporosis.2.The single herbs and bi-combined application can improve the life status of rats,making them more vibrant and stable. After intragastric administration of traditional Chinese medicine for 12 weeks,the rats were dissected at the end of the experiment, finding no obvious visceral lesions,which confirmed the high safety, less side effect in traditional Chinese medicine,proving it can be used as prevention and treatment for osteoporosis.3.The single herbs and bi-combined application could increase the bone mass of femur and lumbar vertebrae in rats, improve the structure characteristics of the bone trabecular, improve the biomechanical properties of femur and lumbar vertebra in ovariectomized osteoporosis rats model.It can also inhibit differentiation and maturation of osteoclast, slow down the bone absorption, reduce the high conversion rate of bone metabolism by reducing the blood serum levels of IL-1, IL-6, ALP levels.It also has similar effect to estrogen, which can regulate blood lipids, induce bone marrow mesenchymal stem cells into osteogenic differentiation, promote osteoblast formation, inhibit the development of osteoporosis.4. The application effect of bi-combined traditional Chinese medicine,which has a synergistic effect,is better than single Chinese herb. |
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