CAJ | 학술논문

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3,6,12,18,24,36,48 h 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,3-6 h 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),18-24 h 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.
배경:중조인골형태발생단백2가이촉진조직공정골혈관화,단시대우기작용우인체세포시적생물학규률불명학.목전대우중조인골형태발생단백2조절인체세포혈관내피생장인자표체적규률국내환미견상관보도.목적:종기인화단백수평관찰비교불동시간점중조인골형태발생단백2유도하인지방간충질간세포혈관내피생장인자적표체.방법:종성인지방조직중분리배양지방간충질간세포,취제3대세포용우실험,분위유도조화대조조.유도조채용종농도위100μg/L 중조인골형태발생단백2유도인지방간충질간세포,분별유도3,6,12,18,24,36,48 h 후수집양본,용 RT-PCR 화 ELISA 분별종기인수평화단백수평검측혈관내피생장인자적표체,병여공백대조조비교.결과여결론:중조인골형태발생단백2조절인지방간충질간세포표체혈관내피생장인자구유시간의뢰성,재불동시간점혈관내피생장인자표체량불동.여공백대조조상비,3-6 h 시간단중조인골형태발생단백2억제혈관내피생장인자표체(P <0.05),18-24 h 시간단중조인골형태발생단백2촉진혈관내피생장인자표체(P <0.05),당이용중조인골형태발생단백2촉진조직공정골혈관화시저량개시간단응당인기특별관주.
@@@@BACKGROUND: Recombinant bone morphogenetic protein-2 can promote tissue engineering bone vascularization, but its biological rules targeting human cel s are not clear. At present, there is in which report on recombinant bone morphogenetic protein-2 adjusts the expression of vascular endothelial growth factor in human cel s. OBJECTIVE: To observe and compare the expression of vascular endothelial growth factor in human adipose-derived mesenchymal stem cel s on gene level and protein level at different time points after induced with human recombinant bone morphogenetic protein-2. METHODS: Adipose-derived mesenchymal stem cel s were separated from adult human adipose tissues and cultured until passage 3, then divided into induced group and control group. The cel s in the induced group were induced by human recombinant bone morphogenetic protein-2 which final concentration was 100 μg/L, then the samples were col ected at 3, 6, 12, 18, 24, 36 and 48 hours after induction. Reverse transcription-PCR and enzyme-linked immunosorbent assay were used to detect vascular endothelial growth factor expression on gene level and protein level, compared with the control group. RESULTS AND CONCLUSION: Human recombinant bone morphogenetic protein-2 adjusted vascular endothelial growth factor expression of adipose mesenchymal stem cel s in a time-dependent manner, and the expression of vascular endothelial growth factor changed at different time points. Compared with the control group, human recombinant bone morphogenetic protein-2 could suppress vascular endothelial growth factor expression at 3-6 hours (P < 0.05), while at 18-24 hours, human recombinant bone morphogenetic protein-2 could promote vascular endothelial growth factor expression (P < 0.05). These two time periods should be paid attention when using human recombinant bone morphogenetic protein-2 to promote tissue engineering bone vascularization.