| Objective To explore the effect of Electroacupuncture on bone density,biomechanical changes and gene and protein expression of Wnt3 a and beta-catenin in ovariectomized rats,and to reveal the mechanism of Electroacupuncture on ovariectomized rats at the molecular level.Methods1.Animal grouping and intervention methods.Sixty SPF female Sprague-Dawley rats,aged about 60 days and weighing 180 ~ 220 g,were randomly divided into two groups: blank group and sham operation group.They were randomly divided into model group,drug group and acupuncture group with 12 rats in each group.Bone mineral density(BMD)and bone mineral content(BMD)were measured to confirm the success of osteoporosis model.After the treatment,the acupuncture group took "Shenshu","Pi Yu","Sanyinjiao","Zusanli",each time electroacupuncture stimulated 30 min and took the same body as the drug group.Distilled water,once a day,continuously treated for 5 days,every 20 days for a course of treatment,for a total of 3 courses of treatment;the drug group rats were gavaged with estradiol valerate solution for the same course of treatment;the blank group was not treated and was fed routinely;the model group was treated with estradiol valerate.Rats in sham-operation group were given distilled water of the same volume as drug group for the same course of treatment,and rats in each group were weighed once a week in order to adjust the dosage.The rats were anesthetized and the femur,tibia and4 th lumbar vertebrae were separated and preserved.2.Indicator detection2.1 total bone mineral density and bone density detection: using double energy X-ray bone density instrument(DXA)to detect the bone density of the left femur and the lumbar spine.2.2 biomechanical detection:(1)three-point bending test: the maximum load and modulus of elasticity of the left femur of BMD were tested by the ags-j series precision electronic universal material testing machine.(2)compression test: the vertebra with bone density was separated out of the fourth lumbar vertebra,placed on the loading platform,and the maximum load and elastic modulus were recorded and calculated.2.3 expression of Wnt3 a and beta-catenin m RNA: RT PCR assay.2.4 expression of Wnt3 a and beta-catenin: immunohistochemical detection.Results1.Biomechanical testing Results of three-point bending test and compression test: compared with sham operation group,the elastic modulus and maximum load of the model group decreased significantly(P<0.05),and the elastic modulus of the drug group and acupuncture group was significantly lower than that of the model group.The maximum load increased significantly(P<0.05),and there was significant difference between the acupuncture group and the drug group compared with the sham operation group(P<0.05).After 13 weeks of feeding,the results of whole body bone mineral density test showed that compared with the blank group,the BMD and bone mineral content of the model group decreased significantly(P<0.05),which proved that the model of osteoporosis was established successfully.The results of lumbar bone mineral density test showed that the BMD of model group was significantly lower than that of blank group and sham-operation group(P<0.05),that of acupuncture group and drug group was significantly higher than that of model group(P<0.05),and that of acupuncture group and drug group was significantly higher than that of sham operation group(P<0.05).The difference was statistically significant(P<0.05).2.Biomechanical testing Results of three-point bending test and compression test: compared with sham operation group,the elastic modulus and maximum load of the model group decreased significantly(P< 0.05),and the elastic modulus of the drug group and acupuncture group was significantly lower than that of the model group.The maximum load increased significantly(P <0.05),and there was significant difference between the acupuncture group and the drug group compared with the sham operation group(P<0.05).3.Wnt3 a and ?-catenin gene expression The lowest expression of Wnt3 a and ?-catenin protein in the model group was significantly lower than that in the control group and sham-operation group(P<0.05),but the expression of Wnt3 a and ?-catenin protein in the model group was significantly higher than that in the model group(P<0.05),and the expression of Wnt3 a and ?-catenin protein in the model group was significantly higher than that in the model group(P<0.05).4.Wnt3 a and ?-catenin protein expression The lowest expression of Wnt3 a and ?-catenin gene in the model group was significantly lower than that in the control group and sham-operation group(P<0.01),but the expression of Wnt3 a and ?-catenin in the model group was significantly higher than that in the model group(P<0.05).Conclusion Acupuncture can effectively increase the bone density of rats,prevent the decrease of bone mass,slow down the degeneration of bone mechanics,strengthen the bone strength,and have a certain therapeutic effect on postmenopausal osteoporosis,and acupuncture treatment of postmenopausal osteoporosis is through regulation of wnt3a(starting factor)and ?-catenin(mediated factor)Protein and gene expression to activate the Wnt signaling pathway,this signal transduction pathway plays an important role in the process of differentiation and proliferation of osteoblast,which regulates the balance of bone remodeling system so as to prevent and treat postmenopausal osteoporosis,thus revealing the mechanism from molecular level. |
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