Genistein Enhances The Anti-osteoporosis Effect Of Vitamin D3 In Vitro

In recent years,osteoporosis has attracted great attention by medical workers and researchers.The amount of bone formed by the proliferation and differentiation of osteoblasts cannot compensate for the bone surface depression formed by osteoclast bone resorption,which the key cause of osteoporosis.Vitamin D3(VD3)and genistein(Gen)have become popular for studying osteoporosis due to their wide source and low price.In this paper,human osteosarcoma MG-63,mouse macrophage RAW264.7,mouse precranial cranial bone cells MC3T3-E1 and RAW264.7 were directly contacted to establish a co-cultivation mode to explore the effect of vitamin D3 combined with genistein on the proliferation and differentiation of osteoblasts and osteoclasts and the possible mechanism.To investigate effects of different concentrations of vitamin D3,genistein and the combination on the proliferation of osteoblast MG-63,the results showed that the combination of vitamin D3 and genistein at a concentration of 6.25 μM significantly increased the proliferation activity of osteoblasts to 1.4 times,and the activity of alkaline phosphatase(ALP)to 1.3 times,which promoted the formation of calcium nodules.It promotes the expression of osteoprotegerin(OPG)and inhibits the expression of receptor Activator for Nuclear Factor-κ B Ligand(RANKL).The mRNA expression ratio of OPG and RANKL is increased to 21 times.Vitamin D3 can promote the differentiation and maturation of osteoclasts,which is at a concentration of 6.25 ~ 50.00 μM,and genistein can reverse this promotion.Vitamin D3 can increase the expression of cathepsin K in osteoclasts to 1.6 times,and the expression of combined genistein cathepsin K to 0.7 times.The results showed that vitamin D3 showed a concentration-dependent enhancement of osteoclast differentiation and maturation.Combined with genistein can significantly inhibit osteoclast differentiation and maturation,and indirectly play an anti-osteoporosis role.In the constructed osteoblast / osteoclast co-culture system,vitamin D3 combined with genistein can enhance the osteoblast ALP activity and the accumulation of differentiated calcium nodules.Vitamin D3 at 50 μM can increase tartrate resistant acid phosphatase(TRAP)activity by 1.13 times,while the combination of genistein can reverse the promotion effect and decrease to 0.8 times.The results show that the co-culture system can enhance the interaction between the cells,and the effect of the drug is consistent with the direction of single cell culture.In summary,this paper studies the effects of vitamin D3 and genistein on the proliferation and differentiation of osteoblasts and osteoclasts in MG-63,RAW264.7-induced osteoclasts and osteoblast / osteoclast co-culture systems.It provides an effective reference for vitamin D3 in the treatment of osteoporosis.