| Objective:1.The rats with kidney yang deficiency type osteoporosis were prepared and gavaged with jiaweiyanghe decoction at different concentrations to detect the bone density and bone tissue morphology of the rats,and to explore the therapeutic effect and dose-ef fect relationship of jiaweiyanghe decoction on them.2.Extract and culture and identify osteoblasts,and explore the effect of jiaweiyanghe decoction at different concentrations on proliferation and mineralization of osteoblasts and the dose-effect relationship.3.The contents of blood calcium,blood phosphorus,E2,ALP,β-catenin,LRP5 and Ru nx2 were detected to explore the effect of jiaweiyanghe decoction on bone metabolis m and the relationship between its therapeutic mechanism and Wnt/β-catenin signaling pathway.Methods:Study 1: select 60 female SD rats,10 in each group were randomly divided into blan kgroup,model group,jiaweiyanghe decoction low dose group,jiaweiyanghe decoction medium dose group,jiaweiyanghe decoction high dose group,positive drug group,e xcept the blank group,the rest of the group are making osteoporosis model with kidn ey Yang deficiency rats,respectively to 0.5% CMC-Na,jiaweiyanghe decoction(0.5g/ml,1 g/ml,2 g/ml),E2V(0.2 mg/kg)to fill the stomach for 2 months.The genera l conditions of the rats in each group were observed,and the rats’ body weight was measured.After the rats were sacrificed,the left femur was taken for bone mineral d ensity(BMD)measurement,and the pathological staining sections of the right femur were used to observe the bone tissue morphology.Study 2: sixteen newborn SD Suckling rats were selected,and 4 of each group were randomly divided into blank group,jiaweiyanghe decoction low dose group,jiaweiyan ghe decoction medium dose group,jiaweiyanghe decoction high dose group.Osteoblas ts were sacrificed,cultured and identified.The prepared blank and the serum with different concentrations of jiaweiyanghe decoction were used to intervene osteoblasts,and the proliferation and mineralization of osteoblasts in each group were observed.Results:1.Femoral BMD of rats: compared with the model group,the femoral BMD of rats i n each treatment group increased to different degrees,except for no significant differe nce in the jiaweiyanghe decoction low dose group(P>0.05),and significant difference in the medium,high-dose and positive drug groups with jiaweiyanghe decoction(P<0.05).2.Bone pathology staining section: compared with the model group,the bone microst ructures of the low-dose jiaweiyanghe decoction group were not significantly improved;the bone trabeculae of the middle-dose and high-dose jiaweiyanghe decoction group were thickened and the spacing was reduced;the bone marrow cavity structure was n ot significantly absent;the bone microstructures of the positive drug group were close to those of the OP rats of the blank group,with abundant and continuous bone trab eculae.3.Detection of bone metabolism related indicators: compared with the model group,s erum calcium level increased significantly in each drug group(P<0.05).The blood ph osphorus level increased to some extent in the low and medium dose groups,but ther e was no significant difference(P>,0.05),while that in the high dose and positive dr ug groups increased significantly(P<0.05).E2 level increased to a certain extent in t he low-dose group of jiaweiyanghe decoction,but there was no significant difference(P>,0.05),while that in the medium dose group,the high-dose group and the positive drug group increased significantly(P<0.05).ALP level decreased significantly in all groups(P<0.05).4.Serum levels of β-catenin,LRP5 and Runx2 were determined: compared with the model group,levels of β-catenin were significantly increased in each group(P<0.05).Although the level of LRP5 increased to a certain extent in the low-dose group of jia weiyanghe decoction,there was no significant difference(P>0.05),while the level in t he medium,high-dose and positive groups of jiaweiyanghe decoction increased signific antly(P<0.05).The level of Runx2 increased to a certain extent in the low-dose grou p of jiaweiyanghe decoction,but there was no significant difference(P>0.05),while t hat in the medium,high-dose and positive groups increased significantly(P<0.05).5.Proliferation of osteoblasts: compared with the blank group,the growth rate of oste oblasts of all drug components was faster(P<0.05),among which the high-dose grouphad the best effect of promoting proliferation in a dose-dependent manner.6.Mineralization of osteoblasts: compared with the blank group,calcium salt deposition of osteoblasts and formation of mineralized nodules were promoted in all the drug a dministration groups,among which the effect of the low-dose group with jiaweiyangh etang was slightly weaker,that of the high-dose group was the best,and that of the medium-dose group was in between.Conclusion:1.The general conditions of rats were observed,femoral BMD of rats was detected,and bone microstructure was observed,proving the successful preparation of rats with kidney yang deficiency type osteoporosis model.2.Jiaweiyanghe decoction can improve the blood calcium,blood phosphorus and E2 le vels in rats with kidney-yang deficiency osteoporosis model,reduce ALP,affect bone metabolism in rats,and promote calcium salt deposition.3.The addition of jiaweiyanghe decoction can improve the serum levels of β-catenin,LRP5 and Runx2 in rats,and activate the Wnt/ β-catenin signaling pathway.4.Jiaweiyanghe decoction can promote the proliferation and mineralization of osteoblast s,which is of great significance to stabilize the balance of bone metabolism.According to the review,jiaweiyanghe decoction can achieve anti-osteoporosis eff ect by increasing bone density,improving bone microstructure and regulating bone me tabolism in a dose-dependent manner,and its effect is related to the activation of Wn t/β-catenin signaling pathway. |
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