| Objective Osteopontin as the breakthrough point,through animal experiments,BushenJianpi Decoction JianGu granule affect to ovariectomized rats bone resorption,to further clarify the role of the prevention and treatment of postmenopausal osteoporosis.Methods1.Animal grouping,Modeling,Treatment and Drawn:Selected the 3 months of clean level female SD rats 81,randomized into 2 groups:the Sham-operation-group 27 and the Ovariectomized(OVX)group 54.The OVX group was removed the ovary.The Sham-operation-group was not removed the ovary.The OVX group were randomly divided into 2 groups:Saline-treat-group and Jiangu granule-treat-group.3 days after operation,once a day,the rats of the Jiangu granule-treat-group were treated with Jiangu granule 5.8g/kg/d,After dissolving the granules with 2ml of saline and feeding,the rats of Sham-operation-group and Saline-treat-group took NS 2ml/d.After 4,8 and 12 weeks of treatment,9 rats were randomly chosen and to be killed In each group.Take the day before the collection of 24 hours urine,Centrifuge to remove the precipitate,and frozen stored at-80? refrigerator,For urinary DPD and urine Cr detection.Take the abdominal aorta blood,and frozen stored at-80?,For the detection of serum TRACP5b.Isolated left femur for bone biomechanics.Separation of Lumbar L3-5 for Bone Mineral Detection.Isolated lumbar L1 and L2 was used for mRNA and the protein determination of OPN,Itgav and Itg?3 in bone tissue.2.Indexes Detection:Bone mineral densities(BMD)were measured by Discovery Wi dual-energy X-ray absorptiometry for the Lumbar vertebra.The maximum load force of bone was measured with IG-A1000N universal material testing machine.Concentrations of serum TRACP-5b and urine DPD were measured by ELIS A.Detection of creatinine content in urine by biochemical Method.The expressions of OPN,Itgav,Itg?3 mRNA in bone tissue were measured by quantitative PCR(Q-PCR).The content of OPN,Itg?v,Itg?3 protein in bone tissue were measured by Western Bloting.Results1.Results of BMD:4 weeks,8 weeks,12 weeks of lumbar vertebra bone mineral density indicators showed that,The BMD of the Saline-treat-group and Jiangu granule-treat-group were lower than those of the Sham-operation-group;There was significant difference between the Saline-treat-group and the Sham-operation-group at 8 weeks and 12 weeks(P<0.01);In the same period the Jiangu granule-treat-group were higher than those in the Saline-treat-group,There was no significant difference between the two groups at 4 weeks(P>0.05),There was significant difference at 8 and 12 weeks(P<0.01).2.Results of bone biological force:4 weeks,8 weeks,12 weeks Bone biomechanical tests showed,The maximal loading force of the Saline-treat-group and Jiangu granule-treat-group were lower than those of the Sham-operation-group;There was significant difference between the Saline-treat-group and the Sham-operation-group(P<0.01);In the same period the Jiangu granule-treat-group were higher than those in the Saline-treat-group,There was no significant difference between the two groups at 4 weeks(P>0.05),There was significant difference at 8 and 12 weeks(P<0.01).3.Results of TRACP-5b:4 weeks,8 weeks,12 weeks serum TRACP-5b levels were measured,The serum TRACP-5b levels of the Saline-treat-group and Jiangu granule-treat-group were lower than those of the Sham-operation-group;There was significant difference between the Saline-treat-group and the Sham-operation-group(P<0.01);In the same period the Jiangu granule-treat-group were lower than those in the Saline-treat-group,There was significant difference(P<0.01).4.Results of DPD/Cr:4 weeks,8 weeks,12 weeks the Urine DPD/Cr were measured,The Urine DPD/Cr of the Saline-treat-group and Jiangu granule-treat-group were lower than those of the Sham-operation-group in the same period;There was significant difference between the Saline-treat-group and the Sham-operation-group(P<0.01);In the same period the Jiangu granule-treat-group were lower than those in the Saline-treat-group,There was no significant difference between the two groups at 4 weeks(P>0.05),There was significant difference at 8 weeks(P<0.01).The difference was statistically significant At 12 weeks.5.Results of OPN,Itg?v,Itg?3 mRNA in bone tissue:4 weeks,8 weeks,12 weeks Real-time quantitative PCR to detect the OPN,Itg?v,Itg?3 mRN A expression,The result displayed that,the Saline-treat-group and Jiangu granule-treat-group were higher than those of the Sham-operation-group,There was significant difference(P<0.01);In the same period the Jiangu granule-treat-group were lower than those in the Saline-treat-group,There was significant difference at 8 and 12 weeks(P<0.01).6.Results of OPN,Itg?v,Itg?3 protein in bone tissue:4 weeks,8 weeks,12 weeks Western Bloting to detect the OPN,Itg?v,Itg?3 mRNA expression,The result displayed that,the Saline-treat-group and Jiangu granule-treat-group were higher than those of the Sham-operation-group,There was significant difference(P<0.01);In the same period the Jiangu granule-treat-group were lower than those in the Saline-treat-group,There was significant difference at 8 and 12 weeks(P<0.01).Conclusion:1.After ovariectomy,the expression of OPN,Itg?v and Itg?3 in bone tissue was increased,and the functional activity of osteoclasts was enhanced,leading to the occurrence of PMOP.2.The expression of OPN,Itg?v and Itg?3 in bone tissue was down-regulated after the treatment of Jiangu granule,inhibit the activity of osteoclasts,and effectively relieve the bone resorption after ovariectomy in rats. |
PDF Full Text Request