SFRP3 Can Promote Osteogenic Differentiation Of Rat Bone Mesenchymal Stem Cells Through Inhibiting Wnt5a Which Triggers Non Classical Wnt Signaling Pathway

Purposes Rat bone marrow mesenchymal stem cells through culturing and identifying,intervened the stimulation of s FRP3 and Wnt5 a factors,detected in the alkaline phosphatase,crimp protein on the cell membrane(Frizzled)and the change of intracellular β-catenin protein content,explored whether s FRP3 can promote the osteogenic differentiation of BMSCs and whether the Wnt signal pathway effected of mesenchymal stem cells into osteoblasts.They provided new theories and methods for the treatment of osteoporosisMethods Bone marrow mononuclear cells were obtained from bone marrow of 4-6 week old rats,and were purified by multiple fluid transfer and cell adhesion characteristics of BMSCs.Rat bone marrow mesenchymal stem cells(MSCs)were cultured on DMEM medium for primary culture and passage culture.Firstly,we used MTT to study the effects of s FRP3 and Wnt5 a on the proliferation of mouse BMSCs.We studied the effects of different concentrations of s FRP3 and Wnt5 a on osteogenic differentiation of mouse BMSCs.s FRP3 and Wnt5 a treated cells with different concentrations as a stimulating factor,and a blank control group,bone induction,on the sixth day of alkaline phosphatase staining,using ELASA assay,alkaline phosphatase in the cytoplasm of the level.Then we study whether s FRP3 promotes rat BMSCs osteogenic differentiation through Wnt signaling pathway.1n M s FRP3,1n M Wnt5 a and 1n M s FRP3+1n MWnt5 a were added to the hole plate,and the establishment of the control group,using ELASA assay,detected the level of alkaline phosphatase detection in the cytoplasm.The level of total protein of each cell extracted and detected Frizzled and the content of β-catenin using Western blot protein.Results 1 s FRP3 and Wnt5 a inhibited the proliferation of mouse BMSCs.With the increase of concentration,the inhibition was enhanced and the number of cells was decreased 2 Compared with the control group,with the increase of the concentration of s FRP3,the level of alkaline phosphatase in the cytoplasm increased gradually,and there was statistical significance between the two groups,P < 0.05 3 Compared with the control group,with the increase of the concentration of Wnt5 a,the level of alkaline phosphatase in the cytoplasm of the cells gradually decreased,and statistical analysis was performed in each group,P < 0.05,the difference was statistically significant 4 The levels of alkaline phosphatase in the cytoplasm of 1n M s FRP3,1n MWnt5 a,1n M s FRP3+1n MWnt5 a and blank control group were detected.s FRP3 group > s FRP3+1n MWnt5 a group > Blank control group> Wnt5 a group,the statistical analysis between the groups,P < 0.05,the difference is statistically significant.5 Frizzled and β-catenin protein in the membrane of 1n M s FRP3,1n MWnt5 a,1n M s FRP3+1n MWnt5 a and blank control group were detected by Western-blot.According to the protein electrophoresis strips from deep to shallow arrangement,the order of s FRP3 group,blank control group,s FRP3+1n MWnt5 a group,Wnt5 a group.Conclusions 1 s FRP3 and Wnt5 a inhibited the proliferation of rat bone marrow mesenchymal stem cells.2 s FRP3 can promote bone marrow mesenchymal stem cells to differentiate into osteoblasts,and promote the expression of alkaline phosphatase,whereas Wnt5 a inhibited rat bone marrow mesenchymal stem cells to differentiate into osteoblasts,and inhibit the expression of alkaline phosphatase.3 s FRP3 inhibited Wnt5 a and promoted the differentiation of rat bone marrow mesenchymal stem cells into osteoblasts and promoted the expression of alkaline phosphatase 4 s FRP3 inhibits Wnt5 a induced non classical Wnt signaling pathway,which promotes the differentiation of rat bone marrow mesenchymal stem cells into osteoblasts.