华西口腔医学杂志

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粪肠球菌脂磷壁酸对人牙周膜成纤维细胞表达Toll样受体2及白细胞介素-1β的影响

靳路远 罗小良 姜艳 谢晓莉   

  1. 中南大学湘雅医院口腔科, 长沙410008
  • 收稿日期:2012-02-25 修回日期:2012-02-25 出版日期:2012-02-01 发布日期:2012-02-01
  • 通讯作者: 谢晓莉,Tel:13973164578
  • 作者简介:靳路远(1985—),女,山西人,硕士
  • 基金资助:

    湖南省科技厅基金资助项目(2010FJ3067)

Study on the expression of Toll like receptor 2 and interleukin-1β induced by Enterococcus faecalis lipoteichoic acid on human periodontal ligament cells

Jin Luyuan, Luo Xiaoliang, Jiang Yan, Xie Xiaoli   

  1. Dept. of Stomatology, Xiangya Hospital, Central-south University, Changsha 410008, China
  • Received:2012-02-25 Revised:2012-02-25 Online:2012-02-01 Published:2012-02-01
  • Contact: Xie Xiaoli,Tel:13973164578
  • About author:Jin Luyuan(1985—),女,山西人,硕士

摘要:

目的探讨体外培养的人牙周膜成纤维细胞(HPDLCs)在粪肠球菌脂磷壁酸(LTA)刺激下表达Toll样受体2(TLR2)及白细胞介素-1β(IL-1β)的情况。方法体外分离培养HPDLCs,采用流式细胞术检测0.1、1、10 μg·mL-1粪肠球菌LTA刺激24 h后HPDLCs表面TLR2表达的改变,酶联免疫吸附法检测12、24、48 h后IL-1β的分泌情况,并用相同质量浓度的大肠杆菌内毒素作为对照;用TLR2中和抗体预处理HPDLCs 1 h,观察1 μg·mL-1 LTA刺激24 h后其IL-1β的分泌量。结果LTA可致HPDLCs表面TLR2表达增加(P<0.05);与对照组相比,粪肠球菌LTA可致HPDLCs分 泌IL-1β增加(P<0.05),刺激12 h后可检测到IL-1β,48 h内呈上升趋势。TLR2中和抗体对粪肠球菌LTA诱导HPDLCs 分泌IL-1β无明显封闭作用。结论粪肠球菌LTA可引起HPDLCs表面TLR2表达及IL-1β分泌增加。

关键词: 粪肠球菌, 脂磷壁酸, Toll样受体2, 白细胞介素-1β, 根尖周病

Abstract:

Objective To investigate the expression of Toll like receptor 2(TLR2) and interleukin-1β(IL-1β) of cultured human periodontal ligament cells(HPDLCs) activated by Enterococcus faecalis(E.faecalis) lipoteichoic acid (LTA). Methods HPDLCs that were obtained from the periodontal tissues of healthy humans were maintained in proper condition. Flow cytometry was used to detect the expression of TLR2 on normal HPDLCs and infectious HPDLCs which were incubated with 0.1, 1, 10 μg·mL-1 E.faecalis LTA for 24 h. IL-1β was detected by enzyme linked immunosorbent assay(ELISA) after incubating with LTA of the above concentration for 12, 24 and 48 h or pretreated with TLR2 neutralizing antibody for 1 h and then co-cultured with 1 μg·mL-1 LTA for 24 h. Results E.faecalis LTA promoted the expression of TLR2 in normal HPDLCs. The difference had statistical significance(P<0.05). IL-1β secretion could be detected 12 h after stimulation with LTA and increasingly escalate within 48 h(P<0.05). TLR2 neutralizing antibody had no evident effect on IL-1β generation stimulating by E.faecalis LTA. Conclusion E.faecalis LTA can increase the expression of TLR2 and IL-1β in normal HPDLCs.

Key words: Enterococcus faecalis, lipoteichoic acid, Toll like receptor 2, interleukin-1β, periapical disease