The Study Of Biological Characteristic And Regulatory Mechanism Of RhBMP-2Induced The VEGF Expression Of Human Adipose-derived Stem Cells

Objective：rhBMP-2is ideal build vascularized tissue engineered bone growth factor, thisproject intends to study different induction concentrations and different induction timepoints rhBMP-2regulation of human fat-derived mesenchymal stem cells express VEGFbiological principles and to explore the p38signaling pathwayregulatory role in thisprocess. Methods：cultured adipose mesenchymal stem cells isolated from adult adiposetissue cultured for three generations. RT-PCR and ELISA detected induced concentration(0ng/ml,50ng/ml,100ng/ml) and different induction time points (3h,6h,12h,18h,24h,36h,48h) from the gene and protein levels of rhBMP-2induced servant fat mesenchymalstem cells express VEGF law. Western-blot detection of different induction concentration(0ng/ml,50ng/ml,100ng/ml,200ng/ml) and different induction time points (3h,6h,24h,36h) the level of phosphorylation of the p38signaling pathway study with VEGFexpression amount of correlation. Experiment with different concentrations (5μM,10μM)using RT-PCR and ELISA detection of human adipose mesenchymal stem cell VEGFexpression changes in p38specific blocking agent sb203580.Result：the same point intime when the rhBMP-2induced concentration of100ng/ml VEGF expression highest.3h-6h and18h-24h two peak periods of the VEGF expression induced group and thecontrol group, which3h-6h induction of VEGF expression is lower than the control group(P<0.05) and18h-24h induction group the amount of VEGF expression was higher thanthat in the control group (P<0.05). The amount of the level of phosphorylation of p38signaling pathway with VEGF closely related and both showed similar trends. the p38blockers sb203580can inhibit the expression of VEGF inhibition and the higher theconcentration of the more obvious effects of VEGF expression15μmol/ml group wasalmost completely inhibited. Conclusion:rhBMP-2regulating human adipose mesenchymal stem cells in a dose-dependent and time-dependent expression of VEGF.rhBMP-2optimal induction concentration of100ng/ml3h-6h and18h-24h is the use ofrhBMP-2to promote tissue engineered bone vascularization key point in time. p38signaling pathway is involved in the process of rhBMP-2to promote humanadipose-derived mesenchymal stem cells express VEGF and plays an important role in theregulation.