牙龈卟啉单胞菌脂多糖诱导THP-1细胞分泌IL-1β、TNF—α、IL-6能力及其相关Toll样受体的差异性  

Diversity of Porphyromonas gingivalis lipopolysaccharide induced IL-1β, TNF-α and IL-6 levels in THP-1 cells and the associated Toll-like receptors

作　　者：楼小戟[1] 张迪亚[1] 严杰[2] 李盛来[3] 陈莉丽[1] 

机构地区：[1]浙江大学医学院附属第二医院口腔科,杭州310009 [2]浙江大学紫金港校区医学院病原生物学系 [3]浙江大学医学院附属口腔医院

出　　处：《中华微生物学和免疫学杂志》2008年第8期717-721,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金面上项目（30471888）

摘　　要：目的探讨牙龈卟啉单胞菌（Porphyromonas gingivalis，Pg）脂多糖（LPS）诱导人单核-巨噬样细胞THP-1分泌IL-1β、TNF—α、IL-6能力及相关Toll样受体（TLR）的差异。方法采用酚水法提取赡ATCC33277株脂多糖（Pg-LPS），并用红外光谱法和鲎试验进行鉴定。采用ELISA试剂盒定量检测Pg—LPS作用的THP-1细胞分泌IL-1β、TNF-α和IL-6水平。采用TLR2或TLR4单克隆抗体阻断试验联合ELISA检测，了解Pg-LPS结合靶细胞上Toll样受体的类型。实验中采用大肠杆菌O111：B4株脂多糖（E—LPS）作为对照。结果1μg／ml Pg-LPS作用THP-1细胞，分别于0．5、6和6h检测分泌IL-1β、TNF-α和IL-6的水平明显升高（P〈0．01）；1μg／ml E—LPS作用THP-1细胞，分别于6、24和24h检测分泌IL-1β、TNF-α和IL-6水平亦明显升高（P〈0．01）。Pg—LPS和E—LPS诱生的上述细胞因子最高浓度相近（P〉0．05）。TLR2或TLR4单抗均可有效阻断Pg-LPS作用的THP-1细胞分泌IL-1β或IL-6（P〈0．05），但仅TLR2单抗显示了阻断TNF-α分泌的作用（P〈0．05）。E-LPS诱导THP-1细胞分泌上述3种细胞因子的活性仅可被TLR4单抗所阻断（P〈0．05）。结论Pg-LPS诱导THP-1细胞分泌IL-1β、TNF-α和IL-6活性略高于E—LPS，TLR2而非TLR4是Pg—LPS介导靶细胞分泌上述细胞因子的主要受体。Objective To determine the diversity of Porphyromonas gingivalis lipopolysacchafide （LPS） induced IL-1β, TNF-α and IL-6 levels in THP-1 cells and the associated Toll-like receptors （TLR）. Methods P. gingivalis strain ATCC33277 LPS （ Pg-LPS ） was prepared using phenol-water method and then identified by both infrared spectrometry and limulus test. The levels of IL-1β, TNF-α and IL-6 secreted by THP-1 cells under inducement of Pg-LPS were quantitatively detected using commercial ELISA kits. The bloc- king test using TLR2 or TLR4 monoclonal antibody （McAb） plus the ELISA were used to determine the types of Pg-LPS binding TLR on the surface of target cells. In this study, a commercial LPS from E. coli strain O111：134 （E-LPS） was used as the control. Results When 1 μg/ml of Pg-LPS induced THP-1 cells for 0.5,6 and 6 h, or 1 μg/ml of E-LPS induced for 6, 24 and 24 h, respectively, the levels of TNF-α, IL-1β and IL-6 were increased obviously （P 〈 0. 01 ）. However, the maximal concentration of the three cytokines induced by Pg-LPS were similar to that induced by E-LPS （ P 〉 0.05 ）. TLR2 or TLR4 McAb could block the effects of Pg-LPS inducing THP-1 cells to secrete IL-1β and IL-6 （ P 〈 0.05 ）, whereas only TLR2 McAb displayed the inhibition of TNF-α secretion （ P 〈0.05 ）. On the contrast, only TLR4 McAb showed the effects blocking the three cytokines secretion in the THP-1 cells under inducement of E-LPS （ P 〈 0.05 ）. Conclusion Pg-LPS shows a slight higher activity inducing THP-1 cells to secrete IL-1β, TNF-α and IL-6 than E-LPS. TLR2, but not TLR4, is the major receptor of Pg-LPS on the target cells to mediate the secretion of the three cytokines.

关 键 词：牙龈卟啉单胞菌 脂多糖 THP-1细胞 炎性细胞因子 TOLL样受体 

分 类 号：R781.42[医药卫生—口腔医学]