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IL-1β对人牙髓成纤维细胞中MMP-9影响的体外研究

王娉婷1,佟雪璐1,陈新梅2   

  1. 1. 天津医科大学口腔医院
    2. 四川大学华西口腔医院
  • 收稿日期:2011-04-01 修回日期:2011-09-01 出版日期:2012-02-15 发布日期:2012-02-15
  • 通讯作者: 陈新梅

The Effect of Interleukin-1β on MMP-9 Expression of Human Pulp Cells in Vitro

  • Received:2011-04-01 Revised:2011-09-01 Published:2012-02-15 Online:2012-02-15

摘要: 目的:探讨牙髓炎关键介质IL-1β对人牙髓成纤维细胞中MMP-9表达水平的影响。方法:体外培养人牙髓成纤维细胞,取4-8代细胞接种于六孔板板孔内作为实验样本,实验组细胞样本培养过程中加入IL-1β,对照组不加IL-1β,常规培养。采用链霉素抗生素蛋白-过氧化酶免疫组化法检测并比较实验组和对照组细胞MMP-9免疫组化染色阳性率和染色颗粒分布特点;采用明胶酶谱法检测并比较实验组和对照组细胞MMP-9的明胶酶谱和负染酶带的积累光密度值。结果:免疫组化结果显示,经IL-1β刺激后的人牙髓成纤维细胞中MMP-9的表达增强(P<0.01),染色阳性颗粒主要分布于细胞胞浆中,少量分布于细胞外基质中;明胶酶谱分析显示,实验组MMP-9水平升高,实验组负染酶带积累光密度值在五个检测时间点均高于对照组(P<0.01)。结论:IL-1β能够促进人牙髓成纤维细胞中MMP-9的合成和分泌,从而导致牙髓中细胞外基质的降解。

关键词: 牙髓成纤维细胞, 基质金属蛋白酶, 白细胞介素-1, 免疫组化, 明胶酶谱

Abstract: Objective: The purpose of this study was to investigate the effect of interleukin-1β (IL-1β), the key factor of dental pulp inflammation, on the synthesis and secretion of MMP-9 of human pulp cells in vitro. Methods: Human pulp cells were separated and cultured in vitro. Cells of 4th-8th generation were incubated in 6 wells plates and divided into two groups. The cells in experimental group were treated with IL-1β, while the cells in control group were not. The streptavidin peroxdase conjugated method was applied to investigate immunohistochemical positive stained rate of cells and the distribution of positive stained particles. immunohistochemical The gelatin zymography method was also applied to investigate the expression of MMP-9 in human dental pulp cells with IL-1β or not, and gelatin zymography to examine the production of MMPs by dental pulp cell in course of cultures by the number of integrated optical density. Results: The results of immunohistochemical method and gelatin zymography revealed that MMP-9 were expressed in normal human dental pulp cells, and IL-1β could increase this level significantly(P<0.01). The immunohistochemical positive stained particles were found mainly in the cytoplasm, while a few of them were found in the extra-cellular matrix. Upon treated with IL-1β, the cells demonstrate significantly elevated levels of synthesis and secretion of MMP-9, showing the higher number of integrated optical density(P<0.01). Conclusion: MMP-9 is probably one of the matrix-degrading MMPs in dental pulp. IL-1β could have the potential to stimulate the production of MMP-9 in human pulp cells and thus result in the inflammation of dental pulp.

Key words: pulp cell, matrix metallproteinase, interleukin-1β, immunohistochemical, gelatin zymography