Experiment Study On The Effect Of Hederagenin On Anti Osteoporosis

Osteoporosis(OP)is a kind of bone metabolic disease which occurs frequently in middle-aged and old men and women,especially in postmenopausal women.Recently,the number of people suffering from osteoporosis has reached nearly 7% of the total population in China owing to the serious trend of aging of population.For this reason,it is urgent to search for more effective drugs and potential treatments for osteoporosis.Inhibiting the formation and exercise bone resorption of osteoclasts may prove to be a potential effective method for the treatment of osteoporosis.Hederagenin(Hed)is a kind of Pentacyclic triterpenoids,which is commonly found in many natural plants.It has been reported that Hed and its derivatives have numerous effects,such as antidepressant,anti-inflammatory,anti-cancer and anti-diabetes,and the like.However,it is unclear whether Hed can prevent and cure osteoporosis.Therefore,this study aims to explore whether Hed also has potential therapeutic effect on osteoporosis.In order to explore the effect of Hed on osteoclast differentiation and function,macrophage model RAW264.7 cells are used as the research object,this kind of cell model can differentiate into osteoclasts induced by RANKL.First of all,we conduct an experiment in vitro by culturing RAW264.7 cells to grope for the optimal functional concentration of Hed on RAW264.7 cells,The results showed that Hed has no obvious toxic effect on cells when the concentration of Hed was lower than 100μM.Hence,50 μM Hed was used as the maximum concentration for subsequent experiments.Then we co incubated RANKL with different concentrations of Hed to observe whether Hed could curb RAW264.7 cells differentiated into osteoclasts.Tartrate resistant acid phosphatase(TRAP)staining and F-actin ring formation detection of osteoclasts was conducted to explore the inhibition of Hed on the formation of osteoclasts.The results of cell experiment in vitro demonstrated that Hed could significantly restrain RAW264.7 cells differentiate into osteoclasts and repress the formation of F-actin ring of osteoclasts.For the purpose of further verifying the prevention and treatment effect of Hed on osteoporosis,A kind of mouse model of osteoporosis caused by bilateral ovariectomy(OVX)was constructed.All mice were divided into three groups: shamoperated group(No OVX + normal saline),control group(OVX + normal saline)and administration group(OVX + Hed).The parameters of bone volume,bone quantity,bone thickness and bone separation/spacing of mice was detected by Micro CT and The level of osteoprotegerin(OPG)and RANKL in serum of mice were tested by ELISA.The results showed that Hed could significantly reduce the level of RANKL and increase the level of OPG in serum of OVX mice.In conclusion,Hed can retard the formation of osteoclasts,significantly reduce the level of RANKL and increase the level of OPG in blood.The results of this study provide an important basis for Hed in the treatment of osteoclast mediated osteoporosis in postmenopausal women.