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背景:重组人骨形态发生蛋白2可以促进组织工程骨血管化,但是对于其作用于人体细胞时的生物学规律不明确。目前对于重组人骨形态发生蛋白2调节人体细胞血管内皮生长因子表达的规律国内还未见相关报道。目的:从基因和蛋白水平观察比较不同时间点重组人骨形态发生蛋白2诱导下人脂肪间充质干细胞血管内皮生长因子的表达。方法:从成人脂肪组织中分离培养脂肪间充质干细胞,取第3代细胞用于实验,分为诱导组和对照组。诱导组采用终浓度为100μg/L重组人骨形态发生蛋白2诱导人脂肪间充质干细胞,分别诱导3,6,12,18,24,36,48h后收集样本,用RT-PCR和ELISA分别从基因水平和蛋白水平检测血管内皮生长因子的表达,并与空白对照组比较。结果与结论:重组人骨形态发生蛋白2调节人脂肪间充质干细胞表达血管内皮生长因子具有时间依赖性,在不同时间点血管内皮生长因子表达量不同。与空白对照组相比,3-6h时间段重组人骨形态发生蛋白2抑制血管内皮生长因子表达(P<0.05),18-24h时间段重组人骨形态发生蛋白2促进血管内皮生长因子表达(P<0.05),当利用重组人骨形态发生蛋白2促进组织工程骨血管化时这两个时间段应当引起特别关注。
BACKGROUND: Recombinant human bone morphogenetic protein-2 promotes the vascularization of tissue-engineered bone. However, the biological rules of human bone morphogenetic protein 2 are unclear. At present, no relevant reports on the regulation of the expression of recombinant human bone morphogenetic protein 2 in human vascular endothelial growth factor have been reported in China. OBJECTIVE: To observe the expression of vascular endothelial growth factor (VEGF) induced by recombinant human bone morphogenetic protein-2 in human adipose-derived mesenchymal stem cells at different time points from the gene and protein levels. METHODS: Adipose-derived mesenchymal stem cells were isolated and cultured from adult adipose tissue. The third generation cells were used for the experiment and divided into induction group and control group. Induced human adipose-derived mesenchymal stem cells were induced with recombinant human bone morphogenetic protein-2 at the final concentration of 100μg / L, and were collected at 3, 6, 12, 18, 24, 36 and 48 hours after induced by RT-PCR and ELISA respectively The level of gene and protein level of vascular endothelial growth factor were detected and compared with the blank control group. RESULTS AND CONCLUSION: Recombinant human bone morphogenetic protein-2 regulated the expression of vascular endothelial growth factor in human adipose-derived mesenchymal stem cells in a time-dependent manner. The expression of vascular endothelial growth factor was different at different time points. Compared with the blank control group, recombinant human bone morphogenetic protein 2 inhibited the expression of vascular endothelial growth factor (VEGF) at 3-6h (P <0.05), and recombinant human bone morphogenetic protein 2 enhanced the expression of VEGF at 18-24h (P < 0.05), these two time periods deserve special attention when using recombinant human bone morphogenetic protein 2 to promote tissue engineering bone vascularization.