| Destination: The clinical treatment of bone injury has long been a research focus of scholars. From The proliferation of osteoblasts and Neovascularization plays a crucial role for bone repair and bone regeneration.Currently the view considered that leptin may participate in a variety of metabolic regulation in vivo,such as Bone metabolism.The VEGF can yet promote either Neovascularization, but also the bone growth.In recent years some experiments associated with cancer at home and abroad show that Leptin can increase the expression of VEGF in a dose-dependent manner,but in terms of osteoblasts has not been seen.To observe the effect of leptin on osteoblasts proliferation and vascular endothelial growth factor (VEGF) expression. To provide a theoretical basis for the clinical treatment of bone repair and regeneration after injury.Method: Removing the Calvarias of newborn rats aged 24h ,then Primary osteoblasts were cultured in vitro by enzymatic digestion. It could be identified as osteoblasts by the Alkaline phosphatase (ALP) staining and mineralized nodules (Alizarin red method).OD values in each group are measured By MTT method ;the expression of VEGFmRNA was assessed by RT-PCR.Consequence:1.24h after inoculation of osteoblasts,a large number of cells forms the adherent colnies,it displays fusiform,triangles,polygons and irregular shape.After the Alkaline phosphatase staining, the most of cell membrane and cytoplasm can see the dark brown particles.When the cells were about 18-20 days after the formation of calcium nodules by alizarin red staining,we can see the scattered red granular material.2.The MTT showed that after 24h, OD values in all experimental groups rises with the increasing concentration of leptin;3.The RT-PCR showed that, VEGF mRNA in neonatal rat osteoblasts has a stable expression; Semi-quantitative analysis showed that with the rising of leptin concentrations, VEGFmRNA expression increased gradually.Conclusion: Leptin can promote osteoblast proliferation; VEGFmRNA can express stability in osteoblasts; VEGFmRNA expression could be up-regulated by leptin in a dose-dependent manner. |
