研究两种干细胞体外矿化过程中LIM矿化蛋白1的表达

摘要/Abstract

摘要： 目的　LIM矿化蛋白1(LIM mineralization protein1, LMP1)是一种胞内非分泌型蛋白，广泛存在于各种组织中，它不仅影响骨基质的矿化而且还是成骨细胞分化和骨形成过程中重要的调节因子。本实验通过检测LMP1在牙髓干细胞和骨髓间充质干细胞体外矿化过程中的表达情况，来研究胞内信号转导分子LIM矿化蛋白1对牙髓干细胞和骨髓间充质干细胞体外矿化的作用，为LMP1作为成骨调节因子,影响骨基质矿化方面的研究提供参考。方法　酶消化法体外培养牙髓干细胞，用挑克隆细胞团的方法对牙髓干细胞进行纯化传代培养。密度梯度离心法体外培养骨髓间充质干细胞。两种细胞分别设置实验组和对照组。实验组培养液中加入矿化诱导液，对照组不加矿化诱导液。分别于培养3、5、7、14天检测碱性磷酸酶的活性。培养21天茜素红染色检测矿化结节形成。培养期间，采用荧光定量PCR方法检测LIM矿化蛋白1、骨形态发生蛋白BMP2、牙本质涎磷蛋白DSPP和Ⅰ型胶原蛋白COL1、核心结合因子RUNX2等成骨标志基因的表达，用SPSS 17.0对结果进行统计学分析。结果　培养3、5、7、14天碱性磷酸酶的活性逐渐提高，且实验组碱性磷酸酶的活性显著高于对照组。培养21天实验组可见矿化结节形成。荧光定量PCR检测到实验组及对照组各个基因均表达，且实验组表达显著高于对照组。结论　发现LIM矿化蛋白1与牙髓干细胞和骨髓间充质干细胞体外矿化过程相关，推测LIM矿化蛋白1可能作为检测牙髓干细胞和骨髓间充质干细胞矿化的一个指标。

关键词: 牙髓干细胞, 骨髓间充质干细胞, LIM矿化蛋白1, 骨形态发生蛋-2

Abstract: Objective　 LIM mineralization protein-1 is non-secret protein in the cell, widely existing in various tissues, which not only affects the mineralization of bone matrix but also is the important regulatory factor in the process of osteoblast differentiation and bone formation. This experiment evaluates the effect of intracellular signal transduction molecules LIM mineralization protein 1on dental pulp stem cells in vitro mineralization by detecting the process of mineralization. It can provide the reference for evaluating the LMP-1 as osteogenesis adjustment factor and affecting bone matrix mineralization research. Methods　 Dental pulp stem cells were cultured in vitro，purified and then subcultured . Bone marrow mesenchymal stem cells were cultured in vitro by density gradient centrifugation method. Two kinds of cells were set up respectively into the experimental group and the control group. In the experimental group, the induction fluid was added to the culture medium, but in the control group it was not added. They were detected in 3, 5, 7 and 14 days respectively. Fluorescence quantitative PCR method was used to detect the culture period expression of osteogenic marker genes LIM mineralization protein-1、BMP-2、DSPP、COL-1、RUNX-2. The results were statistically analyzed by SPSS17.0. Results　The activity of alkaline phosphatase increased and the activity of alkaline phosphatase in the experimental group was significantly higher than that in the control group at 5, 3, 7 and 14 days. The formation of mineralized nodules was seen in the experimental group at 21 days. Fluorescent quantitative PCR was used in both the experimental and control groups, and the expression of the experimental group was significantly higher than that of the control group. Conclusions　LIM mineralization protein -1 is associated with the mineralization of dental pulp stem cells in vitro and bone marrow mesenchymal stem cells. It’s speculated that LIM mineralization protein 1 May as the detection of dental pulp stem cells and bone marrow mesenchymal stem cells is an indicator of mineralization.

Key words: DPSC, bone marrow mesenchymal stem cells , LIM mineralization protein-1, BMP-2.

中图分类号:

R393

费腾孙艳艳周妍袁梦桐刘明月史欣胡伟平. 研究两种干细胞体外矿化过程中LIM矿化蛋白1的表达[J]. , 2017, 37(1): 20-24.

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