Effect Of Glucagon Like Peptide-1Analogues On Gene Expression Of Wnt Signaling Pathway-related Factors In Human Osteoblasts

ObjectiveTo investigate the effect of glucagon like peptide-1analogues (Liraglutide) on the proliferation of the human osteoblasts and the mRNA expression of Wnt signaling pathway-related factors, and to investigate the possible mechanism of action on osteoblasts, explore the relationship between glucagon like peptide-1(GLP-1) and bone metabolism, provide new ideas for the GLP-1applied to the treatment of diabetes and osteoporosis.Methods1.The human osteoblastes are culture in vitro. Osteoblast morphology was observed under inverted phase contrast microscope, were performed osteocalcin immune histochemical staining.2.The human osteoblastes are culture in vitro, the effect of different dose of GLP-1(Omol/L、10-7mol/L、10-8mol/L、10-9mol/L)on primary culture human osteoblastes proliferation is assessed by Cell Counting Kit-8(CCK-8) method after24hours.3.The human osteoblastes are culture with different dose of GLP-1analogues(Omol/L、10-7mol/L、10-8mol、10-9mol/L),and the expression of Wnt-3a、 LRP-5、β-catenin mRNA of Wnt signaling pathway in primary culture human osteoblasts is analyzed by real time polymerase chain reaction (Real-time RT-PCR).4.Statistical analysis:data are presented as x±S. All statistical analysis is performed with windows SPSS13.0. The different between the group are determined by One-way ANOVE. The different between random two groups are determined by LSD. Differences are considered significant at a value of P<0.05. Results1.Most of primary culture human osteoblasts were elongated spindle-shaped, star or irregular polygons.And the osteocalcin immune histochemical staining was positive.2.Proliferation of primary culture human osteoblast was promoted by GLP-1(Omol/L、10-7mol/L、10-8mol/L、10-9mol/L) after24hours. GLP-1could promote the proliferation of human osteoblasts, while the proliferation rate of osteoblasts declined along with the concentration increase.The experimental groups and compared with the control groups were considered significant at a value of P<0.05.3.RT-PCR examination revealed:Low concentration of GLP-1(10-9mol/L) could up-regulate the mRNA expression levels of Wnt-3a、LRP-5、β-catenin in human osteoblasts,In contrast,high concentrations of GLP-1(10-7mol/L、10-8mol/L)could down-regulate the mRNA expression levels of Wnt-3a、LRP-5、β-catenin in human osteoblasts, the experimental groups and compared with the control groups were considered significant at a value of P<0.05.Conclusion1.GLP-1can promote the proliferation of human osteoblasts.2.Low concentration of GLP-1could up-regulate the mRNA expression levels of Wnt-3a, LRP-5and β-catenin in human osteoblasts, Wnt signaling pathway may be involved in the regulatory process.3.High concentrations of GLP-1could down-regulate the mRNA expression levels of Wnt-3a, LRP-5and β-catenin in human osteoblasts, it may promote bone formation of osteoblasts through other pathways.