脂多糖对大鼠牙髓细胞ALP、BSP、DSPP表达的影响

上海口腔医学 ›› 2014, Vol. 23 ›› Issue (4): 431-435.

脂多糖对大鼠牙髓细胞ALP、BSP、DSPP表达的影响

王艳丽¹, 潘克清², 孙艳², 邓婧²

1.青岛大学医学院,山东青岛 266003;
2.青岛大学附属医院口腔内科,山东青岛 266003

收稿日期:2013-12-06 出版日期:2014-08-20 发布日期:2014-10-20
通讯作者: 邓婧,Tel: 0532-82912065, E-mail:dengjing3333@126.com
作者简介:王艳丽(1987-), 女, 硕士研究生, E-mail:wangyanli123wan@126.com
基金资助:
青岛市科技支撑计划项目(10-3-3-3-6-nsh)

Effect of lipopolysaccharide on the expression of ALP、BSP、DSPP in rat dental pulp cells

WANG Yan-li¹, PAN Ke-qing², SUN Yan², DENG Jing²

1.Qingdao University Medical College. Qingdao 266003;
2.Department of Oral Medicine, Affiliated Hospital of Qingdao University. Qingdao 266003, Shandong Province, China

Received:2013-12-06 Online:2014-08-20 Published:2014-10-20
Supported by:
Supported by Qingdao Municipal Science and Technology Supporting Project(10-3-3-3-6-nsh).

摘要/Abstract

摘要： 目的:研究脂多糖(lipopolysaccharide,LPS)对大鼠牙髓细胞牙本质涎磷蛋白(dentin sialophosphoprotein,DSPP)、骨涎蛋白(bone sialoprotein,BSP)及碱性磷酸酶(alkaline phosphatase,ALP)表达的影响。方法:采用组织块法获得大鼠牙髓细胞,体外常规培养并进行鉴定,以含0.1、1、10、100和10000 ng/mL牙龈卟啉单胞菌(Porphyromonas gingivalis,P.g)的LPS作用牙髓细胞1、3、5 d,用实时定量PCR检测DSPP、ALP、BSP mRNA表达的变化,采用SPSS17.0软件包对数据进行统计学分析。结果:镜下贴壁后的细胞形态多样,多呈成纤维样细胞形态,还有部分多角形细胞,胞质突起。实时定量PCR结果显示,与对照组相比,1、10 ng/mL LPS组大鼠牙髓细胞DSPP、ALP、BSP的mRNA表达增高,100、10000 ng/mL LPS组DSPP、ALP、BSP的mRNA表达均降低;在1、3、5 d时,1、10、100和10000 ng/mL LPS组mRNA表达逐渐减少。0.1 ng/mL LPS对mRNA的表达无显著影响。3种因子呈现相似的表达变化趋势。结论:低剂量P.g LPS能促进牙髓细胞ALP、BSP、DSPP的表达,高剂量时则抑制ALP、BSP、DSPP的表达;随着培养时间的延长,促进作用逐渐减弱,抑制作用逐渐增强。

关键词: 牙髓细胞, 脂多糖, 矿化因子

Abstract: PURPOSE: To study the changes of expression of dentin sialophosphoprotein (DSPP), bone sialoprotein (BSP) and alkaline phosphatase (ALP) in rat dental pulp cells, which were treated with lipopolysaccharide (LPS). METHODS: Rat dental pulp cells were cultured by the method of tissue block in vitro and identified. The cells were treated with Porphyromonas gingivalis (P.g) LPS at concentrations of 0.1, 1, 10, 100 and 10000 ng/mL. The effects of treatment were respectively examined by the expression of DSPP, BSP and ALP in rat dental pulp cells with real time PCR (RT-PCR) after 1, 3, 5 days of culture. The data was statistically analyzed with SPSS17.0 software package. RESULTS: Under phase contrast microscopy, the adherent cells were found to be primarily typical fibroblasts, and part of the polygonal cells with protuberant cytoplasm. RT-PCR showed that rat dental pulp cells expressed higher mRNA of BSP, DSPP and ALP at concentrations of 1 ng/mL and 10 ng/mL, whereas the expression of BSP, DSPP and ALP mRNA were lower at concentrations of 100 ng/mL and 10000 ng/mL. After 1, 3, 5 days treatment, the expression of BSP, DSPP and ALP gradually reduced in the cells treated with 1, 10, 100 and 10000 ng/mL LPS; while they had no significant change at the concentration of 0.1 ng/mL LPS. CONCLUSIONS: Lower concentration of P.g LPS promotes the expression of ALP, BSP and DSPP in rat dental pulp cells, but larger concentration has inhibitory effect. With the increase of incubation time, the promoting effect decreases gradually, while the inhibition effect increases gradually.

Key words: Dental pulp cells, Lipopolysaccharide, Mineralization factor

中图分类号:

R781

王艳丽, 潘克清, 孙艳, 邓婧. 脂多糖对大鼠牙髓细胞ALP、BSP、DSPP表达的影响[J]. 上海口腔医学, 2014, 23(4): 431-435.

WANG Yan-li, PAN Ke-qing, SUN Yan, DENG Jing. Effect of lipopolysaccharide on the expression of ALP、BSP、DSPP in rat dental pulp cells[J]. Shanghai Journal of Stomatology, 2014, 23(4): 431-435.

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