The Experimental Study On The Effects Of Chinese Traditional Medicine And Western Medicines On Bone Biomechanics In Ovariectomized Rats

Objective1.An ideal Osteoporosis rats experimental model will be established to provide an available channel for study on the effects of Chinese traditional medicine and Western medicines on bone biomechanics,bone mineral density(BMD) and content change in hydroxyproline of lumbars in ovariectomized rats,at the same time to offer a good platform for determining the bone density by the bone density apparatus.2.To determine the mineral content in bones by bone density apparatus and analyze the data of the missing situation of bone mineral content in osteoporosis rats,and then confirmed whether the experimental animal models on osteoporosis rat were established.After the success of modeling on osteoporosis,then the continuous drug intervention for 3 months were given to the respective group rats to determine if there's any change in bone biomechanics and bone mineral density of thighbones and lumbars and if there's any content change in hydroxyproline of lumbars.To approach the relationships between bone biomechanism and bone mineral density and hydroxyproline,and the curative effect and mechanism of action for the treatement of Chinese traditional medicine and Western medicines for osteoporosis after all the results above have been analysed,and to provide the theoretical basis for making a choice in drugs for the treatment of postmenopausal osteoporosis.Methods1.preparation of modelSixty three-month female Wistar no mating rats(offered by animal experimental center of Nan Fang University) were randomly divided into six groups:OVX group (Ovariectomized group),SHAM group(sham-operated group),C group(Calcitonin group),T group(Tibolone Tablets group),XG group(Xianling Gubao Capsule group) and CT group(union of Calcitonin and Tibolone Tablets group),ten rats each group. to anesthetize the rats by intraperitoneal injection of 2%Pentobarbital(50mg.kg^-1), then made operation to remove both ovaries and surgical approach was at the middle of the rat back.The rats of SHAM group were only removed the same amount of fat with both ovaries.And then to observe the bite and sup,drinking,activity,changing of the weight and mental state during making the model.2 The Plan of Medicine interventionAfter 3 months of free feeding,determining the bone mineral density between OVX group and SHAM group to make sure the models were established,and then weighed them to prepare for giving accurate dosage of intervening the medicine which has been transformed from human being's.The dosage and direction of the medicine is as follows:Calcitonin,rat 4.5IU.kg^-1.d^-1,im;Tibolone Tablets,rat 0.09 Tablet kg-1.d^-1, Xianling Gubao capsule 0.54kg^-1.d^-1,and Tibolone Tablets should be crushed and then diluted for oral taking.as Xianling Gubao capsule3.Target examinationAfter 3 months of drug intervention,the rats were killed.by anaesthesia.Taking the fifth lumbar vertebrae and the femur specimens to determine bone mineral density of the femur and lumbar of all groups,and to determin the three-point bending of rat femur and the 5th lumbar vertebrae compression and determin content of Hyp in the 5th lumbar vertebrae.4.Statistical methodsAll of the data were expressed with(?)±S and analyzed by SPSS13.0,and the weight data were analyzed by repeated measures ANOVA,and the results of bone mineral density of OVX group and SHAM group compared with two means on t test of independent sampler;Comparing with means of all groups,if they were Satisfied with Levene's Test for Equality of Variances,compared with multitude means on LSD of one-way ANOVA;if they weren't Satisfied with,compared with multitude means on Tambane's T₂ method of Welch test;Correlation analysis can be adopted as to know the relationships between Hyp and the maximum load of the L₅lumbar vertebrae;test standard P＜0.05.Results1.Model circumstances.During making the model,the rats were normal in the bite and sup.Because of hurt caused by anaesthesia and operation,three rats died after 2 days of the operation.At that day 3 rats were added according their weight.No unusual case after the addition.Compared with bone mineral density of the SHAM group and the OVX group,there was a significant difference between lumbar vertebra and femur(t=13.175,P=0.000;t=12.748,P=0.000).The models were successful.2.After 3 months of drug intervention,resultof bone mineral density and biomechanics had been determined2.1 pairwise comparations of BMD of the right femur Compared with the OVX group,differences among the other groups are significant(P＜0.01) except for the C group(P=0.653);Compared with the SHAM group,differences among them are significant(P＜0.01);There are no significant differences among groups of T,C T and XL(P＞0.05).Obviously drugs above except for the Salmon calcitonin can play an significant effect on inhibition to the bone mass losing of the right femur,while the improvement didn't approach to the normal level.2.2 pairwise comparations of BMD of the lumbar vertebraeCompared with the OVX group,differences were significant among the other groups except for the C group(P=0.557);Compared with the SHAM group,differences were significant in all groups(P＜0.01);There were no significant difference among groups of T and XL(P＞0.05).but the results of C T group was obviously better than T group or XL group.This did mean that the treatment effect of the union of Calcitonin and Tibolone tablets on osteoporosis was better orther methods.2.3 pairwise comparations of the maximum load of the right femurCompared with the OVX group,differences were significant in all groups (P＜0.01);Compared with the SHAM group,differences were also significant in all groups(P＜0.01);Althrough there's no obvious difference among groups X L,T and C T(P＞0.05),significance was obvious when they compared with group C(P＜0.01).That's to say.compared with Salmon calcitonin,improvement of biomechanics of the right femur was more significant when Tibolone tablets,Xianling Gubao capsules and the union of Calcitonin and Tibolone Tablets each were used in osteoporosis rats.And they three played equivalent effects.2.4 pairwise comparations of the maximum load in the 5th Lumbar vertebrae. Compared with the OVX group,differences were significant in each group(P＜0.01); Compared with the SHAM group,differences were also significant(P＜0.01); Althrough there's no obvious difference among groups of C,T and XL(P＞0.05),but significances were obvious when they compared with CT group(P＜0.05).That's to say.Tibolone tablets,Xianling Gubao capsules and Salmon calcitonin can improve the Lumbar vertebrae biomechanics of Osteoporosis rats significantly.But the group of union of Tibolone tablets and Salmon Calcitonin was more efficient.than orther groups.3.The results of determination for content of hydroxyproline in the 5th lumbar The results of determination for content of hydroxyproline in the 5th lumbar and the lumbar maximum load of all rats were inconsistent with the normal distribution.we adopted Spearman as Correlation Coefficients to analyze data.(r=0.964,P＜0.01).The results of a high degree of linear correlation were shown between Hyp and the lumbar maximum load.Conclusions1.Bilateral oophorectomy is an ideal way of making model for primary osteoporosis. The rats except for SHAM group simulated the adverse effect because of the descending of the estrogen after menopause.It supplied an excellent model for The experimental study on the effects of Chinese traditional medicine and Western medicines on bone biomechanics in ovariectomized rats2.Osteoporosis caused by ovariectomy not only caused decline in bone mineral density but also in collagen content and bone quality.There's obvious correlation between the decline in bone collagen content and decrease in biomechanical bone strength,and osteoporotic fracture has some correlation with the decline of bone collagen content.Tibolone tablets,Xianling Gubao capsules and Salmon calcitonin can increase bone collagen content,and also provide a framework for bone mineralization in the core. 3.Calcitonin usually cannot increase BMD effectively,but Obviously improve bone quality.4.Xianling Gubao capsules could effectively increase BMD and improve bone biomechanics,reveal that Xianling Gubao capsules deserves more widely appliation in the future.5.Tibolone tablets,Xianling Gubao capsules and Salmon calcitonin can improve osteoporotic biomechanical properties of rats obviously,thereby reducing osteoporotic fracture risk.The union of Tibolone tablets and Sahnon Calcitonin can be more efficient.