Effect Of Bushen Jianpi Huoxue Recipe On Intervention Of SFRP1 On Osteoblast Proliferation And Osteogenic Differentiation

ObjectiveTo investigate the effect of Bushen Jianpi Huoxue Decoction on osteoblasts overexpressing SFRP1 and silencing SFRP1 transfected with adenovirus,and observing cell proliferation,ALP activity and expression of bone morphogenetic proteins such as RUNX2,BMP2,OPN and ERRα.Provide new ideas and methods in the adjuvant treatment of osteoporosis and molecular biological mechanisms.MethodsThe recombinant adenoviral vector was over-expressed and silenced by sFRP1,and transfected into rat osteoblast cell line UMR106,which was divided into empty adenovirus group,overexpressed SFRP1 group and silenced SFRP1 group.According to the prepared serum of Bushen Jianpi Huoxue Recipe and normal saline blank,different interventions were performed according to the group,and the proliferation,ALP activity and expression of bone morphogenetic proteins such as RUNX2,BMP2,OPN and ERRα were observed.Result①Changes in cell viability of each groupIn the empty adenovirus group,compared with the blank serum,the cell viability of UMR106 cells was significantly increased at 24h,48h,72h after the intervention of Bushen Jianpi Huoxue Decoction serum(P<0.05).Compared with the blank serum+empty adenovirus group,the activity of UMR106 cells in the blank serum+SFRP1 silencing group was significantly increased at 24h,48h,72h(P<0.05),while the UMR106 cells in the blank serum+SFRP1 overexpression group were 24h,48h.The activity at 72h was significantly increased(P<0.05).Compared with the blank serum+SFRP1 silencing group,the activity of UMR106 cells at 24h,48h,72h was significantly increased(P<0.05).Compared with the blank serum+SFRP1 overexpression group,the activity of UMR106 cells in the drug-containing serum+SFRP1 overexpression group was significantly increased at 24h,48h,72h(P<0.05).②Changes in ALP activity in each groupIn the empty adenovirus group,compared with the blank serum,the ALP activity of UMR106 cells was significantly up-regulated after the serum containing Bushen Jianpi Huoxue Recipe(P<0.05).Compared with the blank serum+ empty adenovirus group,the ALP activity of UMR106 cells in the blank serum+SFRP1 silenced group was significantly up-regulated(P<0.05),while the ALP activity of UMR106 cells in the blank serum+SFRP1 overexpression group was significantly down-regulated(P<0.05).).Compared with the blank serum+SFRP1 silencing group,the ALP activity of UMR106 cells was significantly up-regulated in the serum-containing serum+SFRP1 group(P<0.05).Compared with the blank serum+ SFRP1 overexpression group,the ALP activity of UMR106 cells in the drug-eontaining serum+SFRP1 overexpression group was significantly up-regulated(P<0.05).③Changes in bone formation related proteins in each groupIn the Western blot analysis,the expression of RUNX2,BMP2,OPN and ERR a protein in UMR106 cells was significantly up-regulated in the empty adenovirus group compared with the blank serum(P<0.05).Compared with the blank serum+empty adenovirus group,the expression of RUNX2,BMP2,OPN and ERR a protein in UMR106 cells was significantly up-regulated in the blank serum+SFRP1 silencing group(P<0.05),while the UMR106 cells in the blank serum+SFRP1 overexpression group were RUNX2.The expression of BMP2,OPN and ERR a protein was significantly down-regulated(P<0.05).Compared with the blank serum+SFRP1 silencing group,the expression of RUNX2,BMP2,OPN and ERR a protein in UMR106 cells was significantly up-regulated(P<0.05).Compared with the blank serum+SFRP1 overexpression group,the expression of RUNX2,BMP2,OPN and ERR a protein in UMR106 cells was significantly up-regulated(P<0.05).Conclusion:Overexpression of sFRP1 can decrease the proliferation activity of UMR106 cells,decrease the activity of ALP,and down-regulate the expression of bone morphogenetic proteins such as RUNX2,BMP-2,OPN and ERRα.Silencing sFRP1 and Bushen Jianpi Huoxue Recipe can increase the proliferation activity of UMR106 cells,increase the activity of ALP and up-regulate the expression of bone morphogenetic proteins such as RUNX2,BMP2,OPN and ERR a,and the effect of the two interventions is more significant.Bushen Jianpi Huoxue Recipe can inhibit the overexpression of sFRP1 to a certain extent,and SFRP1 is not the only one in which SFRP1 is regulated by Bushen Jianpi Huoxue Recipe to regulate osteoblast metabolism,promote osteogenic differentiation and promote bone formation protein expression.Targets,there are likely to be other targets that together promote regulation of osteoblast metabolism.