双向电泳分析维生素D对牙髓细胞分化的影响.docVIP

KKME专业医学搜索引擎/ 双向电泳分析维生素D对牙髓细胞分化的影响 双向电泳分析维生素D对牙髓细胞分化的影响 作者：聂敏　边专　樊明文　赵心臣　Mikael Wendel 　　［摘要］ 目的：用双向电泳分析牙髓细胞分化过程中产生或分泌的新蛋白质。方法：在体外培养牙髓细胞，给予一定时间的1,25-双羟维生素D3，促使牙髓细胞分化，结合双向电泳技术分析细胞分化前后蛋白质合成、分泌的变化。结果：pH4-8 和相对分子量MW 20~90 kD范围的蛋白斑点分布较多。双向电泳凝胶图像分析软件ImageMaster 2D Platium比较分析，给予维生素D孵育24 h后，检测到77个新的蛋白点，同时有17个蛋白点消失。结论：维生素D促使牙髓细胞分化，并产生或分泌了一些新的蛋白质，维生素D和这些蛋白质对牙齿的发育、矿化的过程起调节作用。 　　［关键词］ 双向电泳 维生素D 牙髓细胞 发育 　　Studies of New Proteins Involved in Development of Rat Dental Pulp Cells. 　NIE Min, BIAN Zhuan, FAN Ming-wen, et al. School of Stomatology, Wuhan University, Wuhan 430079 　　［Abstract］Objective: The purpose of this paper is to study new proteins produced or secreated by rat dental pulp cells in the course of development. Methods: Isolated of rat dental pulp cells were cultured in vitro. Two groups were incubated with vitamin D for 3h and 24h respectively and one group was set as control. Two-dimensional electrophoresis was used to analysis the proteins of cell culture supernatant. Results: The differential protein expression analysis found that 77 protein spots were specifically detected, and 17 protein spots disappeared in the group treated with Vit D for 24 h. Most spots were focused in the area of pH4-8 and 20-90 kD. Conclusion: It is concluded that Vit D may play a role in the development of rat dental pulp cells, which produce some new proteins certified by 2-D electrophoresis. It is suggested that some new proteins have been produced or secreted by rat dental pulp cells in the course of development. ［Key words］ 2-D electrophoresis Vitamin D Dental pulp cells Development 双向电泳第一相即等电聚焦凝胶电泳，依据蛋白质分子的等电点进行分离：在含有载体两性电解质形成的一个连续而稳定的线性pH梯度中，如果偏离蛋白质的等电点，带有电荷的蛋白质分子可以在电场中移动，一直到蛋白质迁移到其等电点位置时，其净电荷为零，在电场中不再移动。双向电泳第二相即梯度SDS电泳，依据蛋白质的分子量进行分离。最终，样品经过电荷和质量两次分离后，得到点状电泳结果，分离准确精细。所以，双向电泳是蛋白质组学的重要研究方法之一。随着固定pH梯度胶条的出现，双向电泳(2 -DE) 已经成为蛋白质组研究中的核心技术之一； 双向电泳/质谱蛋白质组解决方案能够在整体水平上，大规模、快速地鉴定复杂蛋白样品［1］。 牙齿发育过程中，基因表达的调控受到多种激素的调节， 如转录因子、维生素D等［2］。成骨细胞、成牙本质细胞、成牙