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培养的人牙髓细胞中LIM矿化蛋白1及其相关分子的半定量逆转录聚合酶链反应分析

Semi-quantitative RT-PCR analysis of LIM mineralization protein 1 and its associated molecules in cultured human dental pulp cells.

作者信息

Zhang Qi, Wang Xiaoying, Chen Zhuo, Liu Gang, Chen Zhi

机构信息

Key Laboratory for Oral Biomedical Engineering of Ministry of Education and Department of Cariology & Endodontics, Luoyu Road 237, School and Hospital of Stomatology, Wuhan University, Wuhan, Hubei Province 430079, PR China.

出版信息

Arch Oral Biol. 2007 Aug;52(8):720-6. doi: 10.1016/j.archoralbio.2007.02.005. Epub 2007 Mar 21.

Abstract

OBJECTIVE

LIM mineralization protein 1 (LMP-1), an intracellular signaling molecule, regulates osteoblast differentiation and maturation, as well as bone formation. However, the role of LMP-1 in the differentiation of human dental pulp cells and formation of dentin has not been determined. The study was to investigate the expression of LMP-1, the related proteins, such as bone morphogenetic proteins 2, 6 and 7 (BMP-2, BMP-6 and BMP-7), and core binding factor alpha 1 (Cbfa1) during the differentiation of cultured human dental pulp cells and the formation of mineralized nodules.

DESIGN

Differentiation of human dental pulp cells was induced by dexamethasone, asorbic acid and beta-glycerophosphate. The formation of mineralized nodules, was determined by Von Kossa staining and immunocytochemistry detection of dentin sialoprotein. Expression of LMP-1, the related proteins and the differentiation marker alkaline phosphatase (ALP) was analysed by reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTS

The expression of LMP-1, BMP-2, BMP-6, BMP-7 and Cbfa1 was significantly increased in the process of dental pulp cells differentiation and the formation of mineralized nodules, while the pattern of the expression was distinct.

CONCLUSIONS

The elevated level of LMP-1, BMPs and Cbfa1 expression indicated they might play a role in the differentiation of human dental pulp cells and the formation of mineralized nodules.

摘要

目的

LIM矿化蛋白1(LMP-1)是一种细胞内信号分子,可调节成骨细胞的分化、成熟以及骨形成。然而,LMP-1在人牙髓细胞分化和牙本质形成中的作用尚未明确。本研究旨在探讨培养的人牙髓细胞分化及矿化结节形成过程中LMP-1、骨形态发生蛋白2、6和7(BMP-2、BMP-6和BMP-7)以及核心结合因子α1(Cbfa1)等相关蛋白的表达情况。

设计

用地塞米松、抗坏血酸和β-甘油磷酸诱导人牙髓细胞分化。通过Von Kossa染色和牙本质涎蛋白的免疫细胞化学检测确定矿化结节的形成。采用逆转录-聚合酶链反应(RT-PCR)分析LMP-1、相关蛋白及分化标志物碱性磷酸酶(ALP)的表达。

结果

牙髓细胞分化及矿化结节形成过程中,LMP-1、BMP-2、BMP-6、BMP-7和Cbfa1的表达显著增加,但其表达模式各异。

结论

LMP-1、骨形态发生蛋白和Cbfa1表达水平升高表明它们可能在人牙髓细胞分化和矿化结节形成中发挥作用。

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