Effects Of Icariin Combined With Ginsenoside Rg1 On Osteogenic Differentiation Of Bone Marrow Mesenchymal Stem Cells

This study was to investigate the effect of icariin and ginsenoside Rg1monomer and their compatibility on osteogenic induction of bone marrow mesenchymal stem cells,and to explore their therapeutic potential in osteoporosis.Cell Counting Kit-8（CCK-8 kit） was used to detect the proliferation of cells at 24h,48h and 72h after drug administration.The effects of icariin and ginsenoside Rg1 alone and different concentrations on C57 mouse bone marrow interstitial were observed.The optimal drug concentration was screened for the proliferation of stem cells;alkaline phosphatase and alizarin red staining were performed 16 and 30 days after osteogenic induction,and the mineralization ability of different treatment groups was observed;RT-PCR and WB were used to detect The expression of bone cell marker proteins BMP2,RUNX2,OPN and OCN was investigated.The osteogenic differentiation of bone marrow mesenchymal stem cells in different treatment groups was investigated.The final concentration of icariin was 0.1μmol·L^-1,1μmol·L^-1,10μmol·L^-1,and the final concentration of ginsenoside Rg1 was 10μmol·L^-1and 20μmol·L^-1,40μmol·L^-1,respectively.After treatment with cells under normal culture conditions,the A value of each experimental group increased compared with the control group at each detection time point（P<0.05）,icariin and The optimal concentration of ginsenoside Rg1 monomer was 1μmol·L^-1and 10μmol·L^-1respectively;icariin 1μmol·L^-1and ginsenoside Rg1 was 110μmol·L^-1combination the A value of the compatibility group was greater than that of the monomer（P<0.05）.The optimal concentration of icariin and ginsenoside Rg1 was icariin 1μmol·L^-1and ginsenoside Rg1 was 110μmol·L^-1combination.After 16 days of osteogenic induction,the expression of alkaline phosphatase and the number of calcium nodules in each group were higher than those in the blank control and classical osteogenic control.The staining degree of icariin and ginsenoside Rg1 was higher than other groups.The staining tendency after 30 days of osteogenic induction was consistent with that of 16d.The degree of staining of icariin and ginsenoside Rg1 was higher than that of other groups.Compared with the control group,the drug-added group was able to significantly up-regulate the expression of osteogenic marker proteins BMP2,OCN,OPN,RUNX2,etc.The expression levels of icariin and ginsenoside Rg1 were higher than those of the monomer.In summary,the combination of icariin and ginsenoside Rg1 can increase the osteogenic capacity of bone marrow mesenchymal stem cells,which may be related to the up-regulation of osteogenic factors to promote the differentiation of bone marrow mesenchymal stem cells into osteoblasts to form new bone cells and inhibit bone marrow.Mesenchymal stem cells are related to differentiation of other cells.And icariin and ginsenoside Rg1 can significantly promote the proliferation of bone marrow mesenchymal stem cells,no obvious toxicity,and have a good prospect for the treatment of osteoporosis.