The Effect Of Wnt/β-catenin Signaling Pathway On Regulation Of The Osteogenic Differentiation Of Bone Marrow Mesenchymal Stem Cells In Rats Of Vaspin

Objective:To observe the effect of vaspin on osteogenic differentiation of bone marrow mesenchymal stem cells in SD rats and the potential mechanismsMethod:1.Bone marrow mesenchymal stem cells from 4 weeks SD male rats were isolated and culutured,then were observed and identified by flow cytometry.2.Experimental groups: control group, experimental group, to join the osteogenic induction liquid and different concentrations of vaspin(10ng/ml,50ng/ml, 100 ng/ml)and100 ng/ml vaspin + DKK1(Wnt/β-catenin signaling pathway inhibitor).After 7days,alkaline phosphatase activity were detected by microplate reader;the m RNA expression of ALP、Runx2 and β-catenin were detected by real time quantitative PCR.3.100ng/ml vaspin and osteogenic induction liquid intervene BMSCs.After21 days,alizarin red stained for mineralized nodules.Results:1.Rat bone marrow mesenchymal stem cells at the third passage were fusiform and grew in clusters,and were positive for CD44、CD90,but negative for CD34.2.The effect of vaspin on osteogenic differentiation of bone marrow mesenchymal stem cells:Alkaline phosphatase activity was increased in a dose-dependent manner,compared with the control group, there were no significant differences in the low concentration group(P > 0.05), but the other two groups was significantly increased(P <0.05); ALP、Runx2m RNA was increased in a dose-dependent manner.Compared with the control group,ALPm RNA was significantly increased in different concentration groups(P<0.05).Runx2 m RNA had no significant differences in the low concentration group,but the other two groups was significantly increased(P<0.05).3.Wnt/β-catenin signaling pathway:β-cateninmRNA was increased in a dose-dependent manner. Compared with the control group, only 100ng/ml vaspin group was significantly increased(P<0.05). After joining the DKK1,compared with the 100ng/ml vaspin,alkaline phosphatase activity and ALP、Runx2、 β-catenin m RNA expression were significantly decreased(P<0.05).4.Compared with the control group,mineralized nodules were brown and increased.Conclusions:The rat bone marrow mesenchymal stem(BMSCs) cells provide experimental model for osteogenic differentiation in vitro.Vaspin can promote osteogenic differentiation of BMSCs in a dose-dependent manner,and raise the Runx2 m RNA expression through Wnt/β-catenin signaling pathway to promote osteogenic differentiation.