Pharmacodynamic Molecular Mechanism Of Fennel Total Flavonoids Based On OPG/RANKL/RANK System In Postmenopausal Osteoporosis Model

Objective:To study the effects of cumin total flavones on postmenopausal osteoporosis models and partial surgical removal of postmenopausal osteoporosis in young and older mice induced by complete castration,and to observe the effect of cumin total flavonoids on the proliferation of osteoblast-like cells Saos-2,the influence of potential method on postmenopausal osteoporosis model in young rats,and the role and characteristics of cumin total flavonoids based on OPG/RANKL/RANK system on the pharmacodynamic molecular mechanism of postmenopausal osteoporosis model were discussed.Method:Experiment 1:The postmenopausal osteoporosis model of young mice was induced by complete castration in surgery.The animals were grouped and anesthetized.Except the sham operation group,the other groups were completely removed from the bilateral ovaries.The sham operation group had the same process but Do not remove the ovaries.Penicillin was given for seven days after modeling,and the drug was started on the eighth day of modeling.The drug was continuously administered for 8 weeks.Animal weight was recorded once a week.Urine was collected within 24 hours of the mice two days before material collection.Detection of Ca,P,CRE in the blood;2 hours after the last administration,whole blood was collected by removing eyeballs,and the supernatant was separated after centrifugation for serum index detection;X-ray detection of changes in bone density of femurs in animals;bone strength of small animals The instrument performs biomechanical（maximum load）detection;kidneys,livers,femurs,uterus,thymus,spleen are separated and the changes in organindexes are detected.HE staining is used to observe the morphological changes of organ tissues,ALP staining and Trap staining The distribution of osteoblasts and osteoclasts in femoral tissue was observed by immunohistochemical method.The content of OPG/RANKL/RANK system related proteins in femoral tissue was observed by immunohistochemical method.Experiment 2:The postmenopausal osteoporosis model of older mice was induced by complete castration in surgery.Animals were grouped and anesthetized.Except for the sham operation group,the other ovaries were completely removed from both groups.Do not remove the ovaries.Penicillin was given for seven days after the model was made,and the drug was started on the eighth day of the model.The drug was continuously administered for 8 weeks,and the animal weight was recorded once a week.After 2 hours of the last dose,whole blood was collected by removing blood from the eyeballs.Serum indicators to be tested;X-ray detection of changes in bone density of femurs in animals;small animal bone strength tester for biomechanical（maximum load）detection;kidney,liver,femur,uterus,thymus,spleen and organ index Changes in morphology of organ tissues were observed by HE staining.Experiment 3:The postmenopausal osteoporosis model in young rats was induced by surgical partial castration.Animals were grouped and anesthetized.Except for the sham operation group,bilateral ovaries were surgically removed in the other groups.Do not remove the ovaries.Penicillin was given for seven days after the model was made,and the drug was started on the eighth day of the model.The drug was continuously administered for 8 weeks,and the animal weight was recorded once a week.After 2 hours of the last dose,whole blood was collected by removing blood from the eyeballs.Serum indicators to be tested;X-ray detection of changes in bone density of femurs in animals;small animal bone strength tester for biomechanical（maximum load）detection;kidney,liver,femur,uterus,thymus,spleen and organ index Changes in morphology of organ tissues were observed by HE staining.Experiment 4:Using the MTT method to study the optimal concentration of cumin total flavonoids to promote the proliferation of osteoblast-like cells Saos-2.Based on this,study the OPG/RANKL/RANK system of cumin total flavones on osteoblast-like cells The effects of indicators and levels of Ca²⁺and ALP were investigated.The effects of cumin total flavones on the proliferation of osteoblast-like cells Saos-2 cells and the OPG/RANKL/RANK system were studied.Results:Experiment 1:Complete castration of young mice can be used to replicate a postmenopausal osteoporosis model.The results show that compared with the sham operation group,the model group can significantly increase the weight of mice,the liver,kidney,femur,uterus,thymus,and spleen indexes are significantly reduced,the tibial biomechanics（maximum load）is significantly reduced,and bone density The change is significantly reduced.Serum OPG,E₂,Ca,TGF-β,IL-10,IL-18levels were significantly reduced,RANKL,RANK,P,AKP,IL-3,IL-7,IL-8 levels were significantly increased,and Ca in urine The level is significantly reduced and the CRE level is significantly increased,OPG levels in femoral tissue were significantly reduced,RANKL and RANK levels were significantly increased,osteoblast distribution was significantly reduced,and osteoclasts were significantly increased in femoral tissue.The volume of each organ of the mouse atrophied,and significant lesions appeared.The experimental results suggest that the surgical castration method will interfere with the formation of Ca by affecting the OPG/RANKL/RANK system to form an osteoporosis model.Compared with the model group,Xianling Gubao Capsule Group and Cumin Total Flavonoid Dose Group can improve the animal body weight reduction trend,improve various organ indexes,and significantly increase tibial biomechanics（maximum load）and bone density.Xianling Gubao capsule group and cumin dosage groups can increase serum OPG,E₂,Ca,TGF-β,IL-10,IL-18levels,and reduce RANKL,RANK,P,AKP,IL-3,IL-7,IL-8 level,significantly increase the level of calcium in urine,significantly reduce the level of CRE,OPG levels in femoral tissues were significantly increased,RANKL and RANK levels were significantly reduced,osteoblast distribution in femoral tissues was significantly increased,and osteoclasts were significantly reduced.Improve the pathological changes of various organs,and play the effect of treating postmenopausal osteoporosis.The experimental results suggest that cumin total flavones can be used to treat postmenopausal osteoporosis by interfering with the OPG/RANKL/RANK system,affecting estrogen secretion,interfering with the expression of inflammatory factors mediated by it,and promoting bone reconstruction.Experiment 2:Complete castration of older mice can be used to replicate the postmenopausal osteoporosis model.The results show that compared with the sham operation group,the model group can significantly increase the weight of mice,the liver,kidney,femur,uterus,thymus,and spleen indexes are significantly reduced,and the femoral bone density and tibia biomechanics（maximum load）are significantly reduced.reduce.Serum OPG,E₂,Ca,SOD,and LEP levels were significantly reduced,and RANKL,RANK,P,ALP,BGP,IL-6,and IL-13 levels were significantly increased.The volume of organs in older mice atrophied,and significant lesions appeared.The experimental results suggest that age factors combined with surgical castration will influence the formation of Ca by affecting the OPG/RANKL/RANK system to form an osteoporosis model.Compared with the model group,the Xianling Gubao Capsule Group and Cumin Total Flavonoid dosage groups can cause animal weight loss,increase organ index,and significantly increase femoral bone density and tibia biomechanics（maximum load）.Increase serum OPG,E₂,Ca,SOD,LEP levels,reduce RANKL,RANK,P,ALP,BGP,IL-6,IL-13 levels,improve pathological changes in various organs,and treat postmenopausal osteoporosis Effect.The experimental results suggest that cumin total flavones can prevent and treat postmenopausal osteoporosis by intervening the OPG/RANKL/RANK system.Experiment 3:Partial castration of young rats can be used to establish a postmenopausal osteoporosis model.The results show that compared with the sham operation group,the model group can significantly increase rat weight,significantly reduce liver,kidney,femur,uterus,thymus,and spleen indexes,significantly reduce tibia biomechanics（maximum load）,and bone density Significantly reduced.Serum OPG,E₂,FSH,Ca,BGP,LEP,IL-4,IFN-γlevels were significantly reduced,and RANKL,RANK,P,TNF-α,M-CSF,IL-1,and TRAF6 levels were significantly increased.The volume of various organs in rats was atrophied,and significant lesions appeared.The experimental results suggest that the surgical partial castration method will interfere with the formation of Ca by affecting the OPG/RANKL/RANK system to form postmenopausal osteoporosis.Compared with the model group,Xianling Gubao Capsule Group and Cumin Total Flavonoid Dose Group can reduce the weight of young rats,increase the organ index at the same time,significantly increase the tibial biomechanics（maximum load）,and make bone density Significantly increased serum OPG,E₂,FSH,Ca,BGP,LEP,IL-4,IFN-γlevels increased,RANKL,RANK,P,TNF-α,M-CSF,IL-1,TRAF6 levels decreased,To improve the pathological changes of various organs,to play the effect of treating postmenopausal osteoporosis.The experimental results suggest that cumin total flavones can prevent and treat postmenopausal osteoporosis by intervening the OPG/RANKL/RANK system.Experiment 4:The total flavones of cumin can significantly increase the proliferation rate of osteoblast Saos-2 by MTT method;the total flavones of cumin can significantly increase the OPG and Ca levels in osteoblast Saos-2,and significantly reduce the Saos-2 RANKL and RANK levels.The results of osteoblast proliferation and cell biochemical index level measurement suggest that Cumin total flavonoids can improve the Ca content of cells through the OPG/RANKL/RANK system and have an important effect on the bone reconstruction process.Conclusion:Cumin total flavonoids can improve the post-menopausal osteoporosis animal model and the surgical part of postmenopausal osteoporosis in young mice and older mice by surgery.Castration-induced postmenopausal osteoporosis in young rat models of body weight,bone density,biomechanics（maximum load）,improvement of OPG/RANKL/RANK system-related serum indicators,urine caused by changes in estrogen levels in animal models Fluid indicators and pathological changes of visceral tissues can improve the distribution of osteoblasts and osteoclasts in femoral tissue.Cumin total flavonoids can promote the proliferation of osteoblast-like cells Saos-2 and improve related indicators and can effectively alleviate postmenopausal osteoporosis.