【摘要】 为探讨成骨分化的主要调节因子Runt相关转录因子2(Runt-related transcription factor 2,Runx2)在白藜芦醇(resveratrol,RSVL)诱导小鼠骨髓间充质干细胞(bone marrow mesenchymal stem cell,BMSCs)向成骨细胞分化中的作用,以及以Runx2为研究起点,通过生物信息学分析方法以表明白藜芦醇诱导BMSCs向成骨分化的可能机制。本研究用不同浓度的RSVL(10-9mol/L-10-7mol/L)作用小鼠骨髓间充质干细胞8 d。实时逆转录-聚合酶链反应及Western印记法检测Runx2的mRNA和蛋白质表达水平。生物信息学方法分析及预测鼠Runx2蛋白的性质及特征。结果表明,与对照组相比,10-9mol/L、10-8mol/L、10-7mol/L白藜芦醇诱导Runx2的mRNA表达水平分别增加6%(P> 0. 05)、30%(P<0. 01)、49%(P<0. 01);与对照组相比,10-9mol/L、10-8mol/L、10-7mol/L白藜芦醇诱导Runx2蛋白表达水平分别增加12%(P> 0. 05)、72%(P <0. 01)、94%(P <0. 01)。由此可见,白藜芦醇可显著上调BMSCs中Runx2的mRNA和蛋白质表达水平(P<0. 01),且10-7mol/L的白藜芦醇诱导作用最强。生物信息学预测得知,Runx2属于碱性不稳定蛋白质;二级结构中无规卷曲占主导;存在102个潜在的磷酸化位点、97个潜在O-糖基化位点和8个N-糖基化位点; Runx2具有利于蛋白质间相互作用的结构特点; P1启动子区潜在的转录因子结合位点共28处。疏松的结构特征、丰富的修饰位点及多种转录因子结合位点为探讨白藜芦醇诱导BMSCs的分子机制提供参考。更多还原

【Abstract】 We aim to investigate the role of the main regulatory factor Runt-associated transcription factor2(Runx2) when bone marrow mesenchymal stem cells(BMSCs) are induced by resveratrol(RSVL)into osteoblastic cells. We take Runx2 as the starting point,and use bioinformatics analysis to elucidate the possible mechanism of RSVL inducing osteogenic differentiation of BMSCs. The mRNA and protein levels of Runx2 were detected by real-time reverse transcription-polymerase chain reaction and Western blotting with different concentrations of RSVL(10-9 mol/L-10-7 mol/L) for 8 days. Bioinformatics was used to analyze and predict the properties and characteristics of mouse Runx2 protein. As a result,compared with the control group,the mRNA level of Runx2 induced by 10-9 mol/L,10-8 mol/L,10-7 mol/L RSVL increased by 6%(P > 0. 05),30%(P < 0. 01) and 49%(P < 0. 01),respectively.Compared with the control group,the level of Runx2 proteins by 10-9 mol/L,10-8 mol/L,10-7 mol/L RSVL increased by 12%(P> 0. 05),72%(P < 0. 01),and 94%(P < 0. 01). In summary,RSVL could significantly up-regulate the mRNA expression and protein levels of Runx2 in BMSCs(P< 0. 01),and the induction of RSVL by 10-7 mol/L was the most significant. Bioinformatics prediction analysis shows that Runx2 is an alkaline unstable protein; random crimping is dominant in secondary structures;There were 102 potential phosphorylation sites, 97 potential O-glycosylation sites and eight Nglycosylation sites; Runx2 has structural characteristics conductive to protein-protein interaction; There are 28 potential transcription factor binding sites in the P1 promoter region. The loose structural characteristics,abundant modification sites and multiple transcription factor binding sites provide a reference for exploring the molecular mechanism of RSVL-induced BMSCs.更多还原