上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

牙龈卟啉单胞菌脂多糖及荚膜多糖诱导THP-1细胞分泌IL-8的分析

王依玮，刘大力，束蓉   

  1. 上海交通大学 医学院附属第九人民医院牙周病科，上海市口腔医学重点实验室，上海 200011
  • 出版日期:2017-04-28 发布日期:2017-05-04
  • 通讯作者: 束蓉，电子信箱：shurong1977@163.com。
  • 作者简介:王依玮（1988—），女，住院医师，硕士；电子信箱：wangyiwei880110@163.com。
  • 基金资助:

    上海市科学技术委员会基础研究重点项目（12JC1407502）；上海市科学技术委员会医学重点项目（13411951300）；上海高校高峰高原学科建设项目

Analysis of IL-8 secretion in THP-1 cells stimulated by lipopolysaccharide and capsular polyssacharide extracted from Porphyromonas gingivalis
 

WANG Yi-wei, LIU Da-li, SHU Rong   

  1. Department of Periodontology, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine; Shanghai Key Laboratory of Stomatology, Shanghai 200011, China
  • Online:2017-04-28 Published:2017-05-04
  • Supported by:

    Key Fundamental Research Program of Science and Technology Commission of Shanghai Municipality, 12JC1407502; Key Medical Project of Science and Technology Commission of Shanghai Municipality, 13411951300; Shanghai Summit & Plateau Disciplines

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摘要：

目的 ·分离牙龈卟啉单胞菌（Pg）脂多糖（LPS）及荚膜多糖（CPS），分析两者体外诱导人单核细胞系THP-1细胞分泌IL-8的能力。方法 ·采用凝胶过滤色谱法分离已获得的Pg ATCC33277及SJD4多糖混合物，并作用于THP-1细胞。采用ELISA法检测THP-1细胞在不同菌株LPS和CPS的不同浓度及不同作用时间下，其上清液中IL-8的含量。结果 ·成功分离获得ATCC33277及SJD4的LPS及CPS，其相对分子质量分别为20 000～50 000及150 000～500 000；分离所得LPS及CPS均具有体外诱导THP-1细胞分泌IL-8的能力，且存在时间-剂量效应；在0.1 μg/mL浓度下，模式菌株ATCC33277的LPS诱导能力均显著高于其CPS（均P<0.05），而临床菌株SJD4在作用24 h时其CPS的诱导能力显著高于LPS（P<0.05）。结论 · CPS作为Pg多糖提取物的组成成分，可单独体外诱导THP-1细胞分泌IL-8，且存在时间-剂量效应。

关键词: 牙龈卟啉单胞菌, 脂多糖, 荚膜多糖, THP-1细胞, 白细胞介素-8

Abstract:

Objective · To isolate lipopolysaccharide (LPS) and capsular polysaccharide (CPS) from Porphyromonas gingivalis (Pg) and analyze their ability to induce THP-1 cells to secrete interleukin 8 (IL-8) in vitro. Methods · LPS and CPS were isolated from the polysaccharides by gel filtration chromatography, which had already been extracted from Pg ATCC33277 and SJD4. The isolated LPS and CPS from different strains were used to stimulate THP-1 cells in different doses and for different periods. The level of IL-8 in the cell culture suspension was measured by ELISA. Results · LPS and CPS were successfully separated from the polysaccharides in Pg ATCC33277 and SJD4. The relative molecular masses of LPS and CPS were 20 000–50 000 and 150 000–500 000, respectively. LPS and CPS could induce THP-1 cells to produce IL-8 in vitro in a time- and dose-dependent manner. The effect of LPS from ATCC33277 on secretion of IL-8 was greater than that of CPS with the dose of 0.1 μg/mL (P<0.05). However, the difference could only be found at 24 h in SJD4, and the effect of CPS was greater (P<0.05). Conclusion · As a component of polysaccharides extracted from Pg, CPS can induce THP-1 cells to secrete IL-8 in vitro in a time- and dose-dependent manner.

Key words: Porphyromonas gingivalis, lipopolyssacharide, capsular polyssacharide, THP-1 cell, IL-8