| In recent years,the disease of osteoporosis has become more serious.The World Health Organization has listed osteoporosis as one of the three major diseases of middle-aged and elderly people.Traditional Chinese medicine believes that osteoporosis belongs to the category of "bone callus".There are many reasons for this disease,such as aging,degenerative changes in organ physiological function,decreased secretion of sex hormones,endocrine secretions including thyroxine,insulin,insulin-like growth factor(IGF),and nutrients including protein,lack of vitamin D and osteoporosis caused by kidney function damage.As a traditional medicinal material,antler has been found that velvet antler has obvious calcium supplementation and can improve osteoporosis in rats.Therefore,the study tried to use the sika deer antler as the main raw material,and it was compatible with traditional Chinese medicines with the same roots as Pueraria lobata,psoralen and cucumber seeds.The animal experiment was used to replicate the osteoporosis model to explore the effect of compound antler capsule on dexamethasone combined with low calcium feed.The role of osteoporosis was to provide new ideas and theoretical basis for the in-depth research and application of traditional Chinese medicine in pharmacological action of bone metabolism.Sixty 12-week-old clean female SD rats were rando mLy divided into 5 groups.: control group(Con),model group(Model),calcium gluconate group(CG),Compound antler low group(APPC200),Compound antler high group(APPC400),12 per group.The control group was given 1.2 mL of distilled water per day,the model group was given normal saline by intragastric administration,the calcium gluconate group was given 0.6 mL of calcium gluconate per day.The APPC group was intragastrically administered with different doses of compound velvet capsules,which were 200mg/kg and 400mg/kg respectively.After continuous gavage for 10 weeks,except for the control group,the rats in the other groups changed the normal feed to low-calcium feed from the fifth week,and intramuscular injection of dexamethasone 1mg/kg/3d.After the end of the experiment,the rats were anesthetized with ether,blood was taken from the abdominal artery,and the weight of the femur and tibia were recorded.Serum for calcium(Ca),phosphorus(P),alkaline phosphatase(ALP),estradiol(E2),osteocalcin(OCN),tartrate-resistant acid phosphatase(TRAP),calcitonin(CT)Indicator measurement by ELISA.Soft tissue peeling and smearing on the leg bones of the rats.The right femur is used as a measure of bone density.Determination of hydroxyproline in the right tibia.The right leg bone was used as a measure of ash,calcium and phosphorus content.The left tibia was used for the determination of hematoxylin-eosin staining(HE),Masson staining,and immunohistochemistry(IHC),and the left femur of the rats was tested by PCR.The results of the experiment showed that the 3 months old SD rats were injected with dexamethasone(1mg/kg/3d)combined with 6 weeks of low calcium feed to successfully construct the rat osteoporosis model.Compared with the Con group,the weight of the Model group showed a downward trend.The drinking water in the Model group was higher than that in the Con group,the bone density decreased significantly(P<0.05),the dry weight,ash weight,and ashes/dry weight were all decreased(P>0.05),serum calcium was significantly increased(P<0.05),serum phosphorus was significantly decreased(P<0.05),bone microstructure was destroyed,a large number of fat cells were produced,and trabecular bone had been broken.The experimental results of SD rats in each group showed that the APPC400 group could significantly increase the bone mineral density of the rats(P<0.05),and the bone density of the APPC group was higher than that of the CG group.The ash weight/dry weight of the Model group was lower than that of the APPC group and the CG group,but the difference was not significant(P>0.05).The contents of bone calcium and phosphorus in APPC group and CG group were significantly higher than those in Model group(P<0.05),and the contents of bone calcium and phosphorus in APPC group were higher than those in CG group.The ELISA analysis of serum Ca,P,ALP,E2,OCN,TRAP and CT in each group showed that the levels of OCN,E2,P and CT in the serum of APPC group and CG group were significantly increased(P<0.05).Ca,ALP,and TRAP were significantly lower(P<0.05),but the results of each serum index in the APPC group were better than those in the CG group.Morphological observation of bone tissue showed that the trabecular bone of the bone part in the Model group was significantly reduced and more fat cells appeared,and the collagen fibers were significantly reduced.The APPC group and the CG group had visible trabecular bone,and the fat cells were significantly reduced compared with the Model group.Compared with the CG group,the APPC group had an increased trabecular area and fewer fat cells.The collagen fiber content in the APPC group and the CG group was significantly higher than that in the Model group.The APPC group significantly increased the expression levels of AP-1 and MMP9.Real-time quantitative analysis showed that APPC group and CG group could significantly decrease the expression of RANKL mRNA in rat bone(P<0.05),and significantly increase OPG mRNA expression(P<0.05),and the expression of Runx2 mRNA was significantly increased(P<0.05).Dexamethasone combined with low-calcium feed can successfully establish a rat osteoporosis model.Compound antler capsule has the function of preventing osteoporosis.Its mechanism of action is: blocking the binding of RANKL to RANK by binding of RANKL to OPG.Due to the lack of transcriptional activation signals generated by RANKL-RANK,osteoclast differentiation and maturation are impaired,bone absorption is inhibited,and OPG/RANKL is regulated.By regulating the expression of OPG/RANKL/RANK system and Runx2 gene,osteoblast differentiation and mineralized calcium and phosphorus metabolism,calcium salt deposition and bone formation are promoted to prevent osteoporosis. |
PDF Full Text Request