天津医药

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牙龈卟啉单胞菌脂多糖对人牙龈成纤维细胞炎症刺激作用的研究*

魏丛丛1,蒋少云1,邓嘉胤2   

  1. 1. 天津医科大学口腔医院牙周科
    2. 天津医科大学口腔医院
  • 收稿日期:2012-10-19 修回日期:2013-02-05 出版日期:2013-05-15 发布日期:2013-05-15
  • 通讯作者: 魏丛丛

Inflammatory Stimulation of Porphyromonas Gingivalis Lipopolysaccharide on Human Gingival Fibroblasts

Cong-Cong WEI, ,   

  1. Department of Periodontology, Stomatology Hospital of Tianjin Medical University, Tianjin 300070, China
  • Received:2012-10-19 Revised:2013-02-05 Published:2013-05-15 Online:2013-05-15
  • Contact: Cong-Cong WEI

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摘要： 【摘要】 目的 探讨牙龈卟啉单胞菌(P.g)脂多糖(LPS)刺激人牙龈成纤维细胞(HGFs)分泌炎症细胞因子的作用及其机制。方法 组织块法培养HGFs,用不同浓度(1mg/L、10mg/L)的P.gLPS刺激HGFs后6h和12h,采用酶联免疫法(ELISA)检测细胞分泌肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β),定量real-timePCR检测Toll样受体(TLR)2和TLR4在HGFs中的表达。结果 与非P.gLPS刺激组相比较,P.gLPS刺激HGFs6h和12h后,TLR2的表达和炎症细胞因子TNF-α、IL-1β分泌显著升高,呈浓度依赖性(P<0.05),而且随时间的延长TNF-α、IL-1β分泌明显增加(P<0.05)。TLR4在P.gLPS刺激HGFs6h后的表达明显升高(P<0.05),随着P.gLPS浓度增加其表达量升高更明显,但LPS刺激细胞12h后,TLR4的表达与对照组无明显差别(P>0.05)。结论 P.gLPS能刺激HGFs分泌炎症细胞因子,可能是早期主要通过其表面的TLR2、TLR4来介导炎症细胞因子的分泌,随后主要通过其表面的TLR2来介导炎症反应。

关键词: 人牙龈成纤维细胞, 脂多糖类, 成纤维细胞, 牙龈, Toll样受体2, Toll样受体4, 肿瘤坏死因子α, 白细胞介素1β

Abstract: [Abstract] Objective To explore the effect and mechanism of Porphyromonas gingivalis (P.g) lipopolysaccharide (LPS) on the secretion of inflammatory cytokines in human gingival fibroblasts (HGFs). Methods  HGFs were separated and cultured. Tumor necrosis factor-α and interleukin-1β in the HGFs were examined by enzyme-linked immunosorbent assay at 6 h and 12 h after stimulation with different concentration of LPS (1 μg/ml, 10 μg/ml). Meanwhile, the expressions of TLR2, 4 were examined by real-time PCR. Analysis of variance (ANOVA) statistical analysis was performed with SPSS10.0 software package. Results   Compared with the control group (without LPS stimulation), the expression of TLR2 and secretion of inflammatory cytokine (TNF-α、IL-1β) in HGFs were enhanced obviously at 6 h, 12 h after stimulation of LPS in P.g LPS concentration- and time-dependent manner (P<0.05). The expression of TLR4 was enhanced obviously at 6h after stimulation of P.g LPS in P.g LPS concentration-dependent manner (P<0.05). However, the expression of TLR4 showed no difference between LPS-stimulated group and non-LPS-stimulated group. Conclusion    P.g LPS stimulates HGFs to secrete inflammatory cytokines mainly through TLR2, 4 at early stage. After that, the expression of TLR2 plays an important role in inflammatory reaction induced by p.g LPS in HGFs.

Key words: Human gingival fibroblast, lipopolysaccharides, Fibroblasts, gingival, toll-like receptor 2, Toll-like receptor 4, tumor necrosis factor-alpha, 白细胞介素1β