华西口腔医学杂志

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不同fimA基因型牙龈卟啉单胞菌刺激口腔上皮细胞白细胞介素-8的表达

郭永华1,2, 吴亚菲3, 刘天佳3, 张静宜1, 肖晓蓉1, 赵蕾3   

  1. 1.口腔疾病研究国家重点实验室, 四川大学, 四川成都610041;2.首都医科大学附属北京口腔医院牙周黏膜科, 北京100050;3.四川大学华西口腔医学院口腔内科学教研室, 四川成都6
  • 收稿日期:2008-12-25 修回日期:2008-12-25 出版日期:2008-12-20 发布日期:2008-12-20
  • 通讯作者: 吴亚菲,Tel:028-81801057
  • 作者简介:郭永华(1977-),女,山东人,住院医师,博士
  • 基金资助:

    国家自然科学基金资助项目(30471890)

Interleukin-8 regulations of oral epithelial cells by Porphyromonas gingivalis with different fimA genotypes

GUO Yong-hua1,2, WU Ya-fei3, LIU Tian-jia3, ZHANG Jing-yi1, XIAO Xiao-rong1, ZHAO Lei3   

  1. 1. State Key Laboratory of Oral Diseases, Sichuan University, Chengdu 610041, China;2. Dept. of Periodontics and Oral Medicine, Beijing Stomatological Hospital, Capital University of Medical Science, Beijing 100050, China; 3. Dept. of Oral Medicine, West China College of Stomatology, Sichuan University, Chengdu 610041, China
  • Received:2008-12-25 Revised:2008-12-25 Online:2008-12-20 Published:2008-12-20
  • Contact: WU Ya-fei,Tel:028-81801057

摘要:

目的比较不同fimA基因型牙龈卟啉单胞菌(P.gingivalis)刺激下口腔上皮细胞白细胞介素-8(IL-8)的表达水平,初步探讨P.gingivalis致病性与其fimA基因型的关系。方法P.gingivalis ATCC 33277(Ⅰ型fimA)、W83、47A-1(Ⅳ型fimA)和KB细胞ATCC CCL-17共同孵育24 h,以未受刺激的KB细胞作为对照组,分别在1、3、6、24 h收集细胞和培养上清液。RT-PCR检测KB细胞IL-8 mRNA的动态表达,酶联免疫反应检测培养上清液中IL-8的动态变化。结果2种fimA基因型菌株刺激1 h,KB细胞IL-8 mRNA的表达上调至峰值,高于对照组,3~24 h IL-8 mRNA的表达水平接近对照组;P.gingivalis感染细胞后3~24 h,上清液中的IL-8水平低于对照组,Ⅳ型菌株低于Ⅰ型菌株;IL-8 mRNA及其蛋白表达不完全一致,提示IL-8的表达存在转录后水平的调节。结论fimA基因型与口腔上皮细胞IL-8的表达水平相关,提示P.gingivalis致病性与其fimA基因型相关。

关键词: 牙龈卟啉单胞菌, fimA基因型, 口腔上皮细胞, 白细胞介素-8

Abstract:

Objective The expression of heterogenic virulence properties depends on its clonal diversity. The aim of the study was to investigate the mechanism of interleukin -8(IL -8) regulations of oral epithelial cells by challenge of Porphyromonas gingivalis(P.gingivalis) with different fimA genotypes, discuss the relation between fimA genotype and its pathogenicity. Methods P.gingivalis ATCC 33277(type Ⅰ), W83(type Ⅳ), 47A -1(type Ⅳ) were assessed for their inductions of IL-8 expression in human oral epithelial cells(KB cell line, ATCC CCL- 17). KB cells without stimulation of P.gingivalis were used as control group. IL -8 mRNA expression was de - termined by reverse transcription polymerase chain reaction(RT -PCR) at different time intervals(1, 3, 6, 24 h) following continuous co culture of bacteria with KB cell line, and IL -8 protein levels in culture supernatant was determined by enzyme-linked immunosorbent assay.  esults IL-8 mRNA levels were up-regulated and reached its high peak at 1 h following both genotypes infection, then decreased to base level till 24 h. Attenuation of IL-8 protein levels was down-regulated when KB cell co-cultured with both genotypes for 3 h till 24 h, and type Ⅳ was lower than type Ⅰ. IL-8 and IL-6 mRNA expression were not consistent with their protein levels, which indicated post -transcriptional regulations. Conclusion fimA genotypes of P.gingivalis are related with the effect of IL -8 inductions, which indicates fimA genotype is associated with pathogenesis of P.gingivalis.

Key words: Porphyromonas gingivalis, fimA genotype, oral epithelial cell, interleukin-8