骨髓间充质干细胞诱导成骨细胞模型中活化转录因子4基因的表达  

Expression of activating transcription factor 4 in osteogenic induction of bone marrow mesenchymal stem cells

作　　者：吴丹[1] 李苏伶[1] 王璐[1] 徐凌[1] 向华居 

机构地区：[1]重庆医科大学附属口腔医院,重庆市400016 [2]重庆市巫溪县中医院,重庆市巫溪县405800

出　　处：《中国组织工程研究》2014年第1期21-26,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:研究发现活化转录因子4为近调控成骨细胞分化和功能的活化因子,在成骨细胞分化过程中起着至关重要的作用。目的:探索SD大鼠骨髓间充质干细胞成骨诱导过程中活化转录因子4基因的表达变化及其意义。方法:以全骨髓贴壁法培养SD大鼠骨髓间充质干细胞,传代至第3代细胞时加入成骨诱导剂,在诱导的第0,1,4,7,10,13,16,19天分别采用PT-PCR,Western blot动态监测活化转录因子4基因及蛋白的表达变化,以未进行成骨诱导细胞做对照。结果与结论:RT-PCR结果显示,活化转录因子4 mRNA的表达随着细胞成骨分化程度加剧而升高,16 d达到峰值。Western blot分析结果显示,活化转录因子4蛋白表达量随着骨髓间充质干细胞骨化程度加剧,表达水平亦呈上升趋势,于16 d达到高峰,19 d维持在高水平状态。实验组各时间点活化转录因子4 mRNA和活化转录因子4蛋白表达与对照组相比差异有均有显著性意义(P<0.05)。结果表明诱导成骨细胞中活化转录因子4表达增强,提示活化转录因子4的增强与骨髓间充质干细胞成骨分化能力呈正相关。BACKGROUND：Activating transcription factor 4 is found as an activating factor that can regulate osteogenic differentiation and function, and plays a critical role in the osteogenic differentiation. OBJECTIVE：To investigate the expression and significance of activating transcription factor 4 in the osteogenic differentiation of bone marrow mesenchymal stem cells from Sprague-Dawley rats. METHODS：Bone marrow mesenchymal stem cells from Sprague-Dawley rats were cultured using the whole bone marrow adherence method, and ossification revulsant was added to induce passage 3 cells. cells with no osteogenic induction served as controls. RT-PCR and western blot assay were employed to dynamical y monitor expression of activating transcription factor 4. RESULTS AND CONCLUSION：RT-PCR results showed that activating transcription factor 4 mRNA expression increased with the increasing osteogenic differentiation, and peaked at day 16. Western blot analysis showed that the protein expression of activating transcription factor 4 tended to increase with the increasing osteogenic differentiation, peaked at day 16 and stil maintained at a higher level at day 19. Compared with the uninduced cells, activating transcription factor 4 in the induced cells exhibited a higher expression at mRNA and protein levels （P＆lt;0.05）. These findings indicate that activating transcription factor 4 expression is elevated during osteogenic differentiation, showing a positive correlation with osteogenic ability of bone marrow mesenchymal stem cells.

关 键 词：干细胞 骨髓干细胞 骨髓间充质干细胞 成骨诱导 活化转录因子4 矿化结节 茜素红染色 ACTIVATING transcription factor 4 

分 类 号：R394.2[医药卫生—医学遗传学]