| Objectives:Establish a disease model of osteoporosis(OP)in rats;Treatment with phytoestrogen salvia miltiorrhiza;To investigate the effect of salvia miltiorrhiza combined with calcium carbonate D3 on bone metabolism in ovariectomized osteoporosis rats,and to provide a theoretical basis for the use of phytoestrogens in the treatment of osteoporosis.Methods:1.Establish rat osteoporosis disease model:Twenty 3-month-old SPF-grade healthy female SD rats were subjected to bilateral ovariectomy to construct a postmenopausal SD rat model of osteoporosis.After building 8 w,the pathological changes of femoral metaphyseal bone and the changes of serological parameters(blood calcium,phosphorus,serum alkaline phosphatase,and estradiol levels)in the rat femoral metaphysis were determined.The rats successfully modeled were randomly divided into OP model group,Salvia miltiorrhiza group,Salvia miltiorrhiza+calcium carbonate D3 group,5 rats in each group,and 5 SD female healthy rats of the same batch were randomly selected as normal controls.2.Drug treatment:After the model was successfully established,the drugs were administered in groups according to the treatment conditions.The salvia miltiorrhiza group was given lmg·kg-1 salvia miltiorrhiza injection,the combination group was given 1mg·kg-1 salvia miltiorrhiza injection and calcium carbonate D3 150mg·kg-1,the normal control group and the OP model group were given equal volume of normal saline(0.9%Nacl)once a day for 5 weeks.3.Detection indicators:After 5 weeks of administration,the weight changes of rats in each group were detected,and the contents of serum calcium,serum phosphorus,alkaline phosphatase and estradiol were measured.At the same time,the femoral metaphysis bone tissue of rats was collected for decalcification,paraffin embedding,and HE staining,and histomorphological observation and evaluation were conducted by optical microscope,and the bone trabecular structure and width of each group were observed.At the same time,the decalcified tissue was scanned by electron microscopy to detect the structural changes of femoral trabeculae in each group.Results:1.After the establishment of the osteoporosis disease model,the body weight and serum alkaline phosphatase in rats increased,while the serum estradiol,blood calcium and blood phosphorus decreased compared with normal rats.Compared with the normal control group,the weight and serum alkaline phosphatase in the OP model group were significantly increased(p<0.01).Serum estradiol and serum calcium were significantly decreased(p<0.01),and blood phosphorus was lower than that of normal rats(p<0.05).In the OP model group,pathological staining and scanning electron microscopy showed that the trabecular bone became thinner,the spacing was wider,and fractures occurred to varying degrees.2.Serological indicators:after 5w of drug treatment,compared with the normal control group,serum calcium,serum phosphorus and serum estradiol levels in the OP model group significantly decreased(p<0.01),and serum alkaline phosphatase significantly increased compared with the normal control group(p<0.01).Compared with the OP model group,serum calcium and serum phosphorus in the combination group were significantly increased(p<0.01),serum estradiol was increased compared with the OP model group(p<0.05),and serum alkaline phosphatase was decreased(p<0.05).Blood calcium,blood phosphorus and serum estradiol in salviae miltiorrhiza group increased compared with OP model group(p<0.05),while serum alkaline phosphatase decreased(p<0.05).3.Histopathological changes:Through optical microscope observation and analysis,the bone trabeculae in the OP model group were fractured and became sparse,and the thickness of bone trabeculae became thinner with different degrees of fracture,and there were a large number of fat cells infiltration in the bone marrow cavity.The bone morphology of rats in the salvia miltiorrhiza group was improved compared with the model group,but the infiltration of fat cells in the bone marrow cavity was still observed.Compared with the OP model group,the bone trabecular thickness of the rats in the combined treatment group increased significantly,and the fat cells in the bone marrow cavity decreased.The bone tissue morphology of the rats in the combined treatment group was approximately similar to that in the normal control group.4.Scanning electron microscope:After 5 weeks of salvia miltiorrhiza treatment,the rats in the normal control group had abundant cancellous bone in the distal femur,and the number of bone trabeculae was the largest in each group,with close arrangement and good connectivity,forming a three-dimensional network structure.In the OP model group,the three-dimensional network structure of bone trabeculae was damaged,and the bone trabeculae became thinner.The number of bone trabeculae decreased and the thickness was uneven.The distance between trabeculae increased and the arrangement was irregular.The number of bone trabeculae of rats in the salvia miltiorrhiza group was higher than that in the model group.Compared with the normal control group,the bone trabeculae of the rats in the combined group were slightly less,with more uniform thickness and better preservation of the three-dimensional network structure.Conclusions:1.SD female rats undergoing bilateral ovariectomy can successfully establish a rat osteoporosis disease model,and it is a scientific and feasible modeling method with simple construction,short modeling period,good stability and strong repeatability;2.Salvia miltiorrhiza has therapeutic effect on bone damage in ovariectomized osteoporosis rats,and the combined effect of calcium carbonate D3 treatment on bone tissue repair is better than that of salvia miltiorrhiza alone.3.Salvia miltiorrhizae regulates calcium and phosphorus metabolism,ALP,and exerts Estrogen-like effects have a repairing effect on the structure and number of damaged trabeculae,make bone conversion tend to be balanced,and promote the repair and repair of bone tissue. |
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