| China has gradually entered the aging society,so the health of the elderly people have been paid more and more attention by modern medicine.Osteoporosis is a common,frequently-occurring disease,it seriously threatens the health of the middle-aged and elderly people,resulting in fractures and other complications,in addition to the patient himself caused great pain,but also to the society and family brought a heavy economic and living burden.Osteoporosis is a systemic skeletal disease characterized by low bone mass and microarchitectural deterioration of bone tissue,with a consequent increase in bone fragility and susceptibility to fracture.In China,the overall prevalence of osteoporosis in people over 60 years old was 36%,among which 23%were males and 49%are females.Similar findings have been made in animal medicine.Pet diseases have gradually shifted from infectious diseases to chronic and senile diseases,including bone diseases such as osteoporosis.Osteoporosis in pets was often manifested as fracture,vertebral deformation and bone pain in elderly animals.Epimedium brevicornu was a traditional Chinese medicine.It had the effect of nourishing kidney Yang,strengthening muscles and bones,removing wind and dampness,and had a good effect on osteoporosis in the elderly.Epimedium A was the most abundant active component in epimedium group.Therefore,this study explored the effect of herba epimediensis and its main component Epimedin A on the anti-osteoporosis effect of SD rats and ICR mice.The effect on the proliferation,mineralization and adipogenic differentiation of osteoblasts in vitro,as well as the effect on the bone resorption of primary osteoclasts and the mechanism of action provide experimental basis for the clinical use of icarium and the research and development of new drugs.The experiment consists of five parts:1.The water extracts of 0.5g/kg,1g/kg and 2g/kg of epimedium were used in the rat model of hormone-deficient osteoporosis induced by ovary/testicle removal.Serum bone conversion markers,tibia diameter and bone microstructure were detected.The results showed that compared with the model group,the bone volume,bone area,trabecular number,trabecular thickness,and tibial diameter were significantly increased in the drug administration group(P<0.05,0.01).It is proved that the water extract of epimedium can improve the osteoporosis induced by castration in male and female rats.2.Five flavonoid monomers with the highest content in Epimedium,icariin,baohuoside I,Epimedin A,B and C,were screened in vitro on primary osteoblast and osteoclast to detect the key indicators of osteoblast and osteoclast,ALP and TRACP.The results showed that the five flavonoids promoted the ALP secretion of MSCs respectively(P<0.05,0.01),and there was no significant difference between the two groups.Epimedium A,B and C significantly inhibited the TRACP activity of osteoclasts(P<0.05,0.01).It was proved that icariin,baohuoside I,Epimedium A,B and C can play a certain role in bone protection.Among them,Epimedin A had the strongest bone protective effect,so Epimedin A was selected for subsequent studies.3.5,10,20mg/kg Epimedin A was used to observe the anti-osteoporosis effect of castration-induced ICR in male and female mice.After the administration,mice were sacrificed and serum biochemical,three-point mechanical experiments and bone microstructure detection were conducted.The results showed that 8 weeks after ICR intragastric administration of zhaohopidin A in both male and female mice,the bone mass,number of trabecular bone and bone strength of the mice increased significantly,the trabecular space decreased significantly,and the bone conversion index was significantly improved.The results showed that Epimedium A could improve bone microstructure and increase bone strength in both male and female mice with osteoporosis induced by castration.4.Epimedin A was applied to mesenchymal stem cells from bone marrow of normal mice to observe its effect on osteoblast differentiation,lipogenic differentiation and mineralization ability of mesenchymal stem cells.ELISA and Western Blotting techniques were applied to investigate the molecular mechanism of promoting bone formation.The results showed that epimedium A could promote the differentiation of MSCs cells into osteoblasts and inhibit their lipogenic differentiation,promote the mineralization of osteoblasts and increase the levels of OPG/RANKL and ERK protein phosphorylation in osteoblasts,thus promoting the formation of bone.5.Epimedin A was used in osteoclasts induced by mononuclear cells in bone marrow of normal mice to observe its effect on osteoclast differentiation and bone absorption capacity.Biological methods such as MSD and Western Blotting were applied to study its anti-osteoporosis effect and related molecular mechanisms.The results showed that Epimedin A can inhibit the formation,differentiation and bone absorption of osteoclasts,possibly by regulating the phosphorylation of cjun in the MAPK pathway and AKT protein in the AKT pathway.To sum up,the water extract of Epimedium can improve the osteoporosis induced by castration in both male and female rats.Among the five flavonoids with the highest content in Epimedium,Epimedin A had the strongest comprehensive ability to promote bone formation and inhibit bone absorption.Epimedin A had an anti-osteoporosis effect on both male and female castrated mice,which is related to its promotion of MSCs differentiation into osteoblasts,inhibition of its lipogenic differentiation,and reduction of osteoclast formation and differentiation.Further studies revealed that Epimedin A might play a bone protective role through OPG/RANKL,MAPK and AKT signaling pathways. |
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