酪酸对大鼠成骨细胞系中骨唾液酸蛋白转录的作用

发布时间：2018-05-27 03:06

本文选题：酪酸 + 成骨　；参考：《广东医学》2016年11期

【摘要】：目的研究酪酸调节大鼠成骨细胞系(ROS17/2.8)中骨唾液酸蛋白(BSP)转录的作用机制。方法使用能生成BSP的大鼠成骨细胞系(ROS17/2.8),分为试验组和对照组。试验组首先加入无血清α-基本必须培养基(α-MEM)配制的(10-8~10~(-2))mol/L浓度梯度的酪酸12 h,确定最优刺激浓度为10~(-4)mol/L,然后使用10~(-4)mol/L的酪酸作用于ROS17/2.8细胞3、6、12 h,对照组加入等量不含酪酸的无血清α-MEM。用Northern杂交试验检测BSP mRNA的表达;用瞬时转染方法检测BSP启动子的转录活性变化;用凝胶迁移实验验证酪酸对核蛋白与相关基因区段结合的作用。结果酪酸作用ROS17/2.8细胞12 h后BSP mRNA的表达水平升高。酪酸对BSP基因启动子转录活性的上调作用主要通过启动子p LUC3(-116~+60)中的纤维母细胞生长因子2响应区段(FRE),在凝胶迁移试验中,酪酸促进了FRE片段与相应核蛋白的结合。结论酪酸通过BSP启动子中的FRE片段增强BSP基因的转录,从而在成骨早期增强成骨细胞活性。
[Abstract]:Objective to study the mechanism of tyroic acid regulating the transcription of bone sialic acid protein (BSPs) in rat osteoblast cell line (ROS17 / 2.8). Methods the rat osteoblast cell line, ROS17 / 2.8, which could produce BSP, was divided into two groups: experimental group and control group. The experimental group was treated with serum-free 伪 -essential medium (伪 -MEM) for 12 h with a concentration gradient of 10 ~ (-8) ~ 10 ~ (-10) ~ 10 ~ (-1) O ~ (-1) mol 路L ~ (-1). The optimal concentration of tyroic acid was determined to be 10 ~ (4) 渭 mol / L. Then, the ROS17/2.8 cells were treated with 10~(-4)mol/L for 3 ~ 612 h. The control group was added a serum-free 伪 -MEM without butyric acid. The expression of BSP mRNA was detected by Northern hybridization assay, the transcriptional activity of BSP promoter was detected by transient transfection method, and the effect of casein on the binding of nucleoprotein to related gene regions was verified by gel migration assay. Results the expression of BSP mRNA in ROS17/2.8 cells increased 12 h after treatment with butyric acid. The upregulation of the transcriptional activity of BSP gene promoter by caseic acid was mainly through the FRE2 response region of promoter pLUC3- 116-60. In the gel migration test, casein promoted the binding of FRE fragment to the corresponding nucleoprotein. Conclusion tyroic acid enhances the transcription of BSP gene through the FRE fragment of BSP promoter, thus enhances the activity of osteoblasts in the early stage of osteogenesis.
【作者单位】：天津市口腔医院口腔颌面外科;山西省人民医院口腔颌面外科;日本大学松户齿学部;
【分类号】：R781.4

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