比较牙龈卟啉单胞菌脂多糖诱导THP-1和HL-60细胞分泌IL-1β能力及相关信号通路受体的差异  

Effects of Porphyromonas gingivalis lipopolysaccharide on induction of IL-1β in THP-1 and HL-60 cells and the associated signaling receptors

作　　者：李盛来[1] 张迪亚[2] 严杰[3] 陈莉丽[2] 

机构地区：[1]浙江大学医学院附属口腔医院,杭州310006 [2]浙江大学医学院附属第二医院口腔科 [3]浙江大学医学院病原生物学系

出　　处：《现代口腔医学杂志》2008年第2期176-179,共4页Journal of Modern Stomatology

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脂多糖(Porphyromonas gingivalis,Pg-LPS)诱导人单核巨噬细胞株THP-1和人粒细胞白血病细胞株HL-60分泌白介素-1β(interleukin,IL-1β)能力差异,并了解其相关Toll样受体。方法采用酚水法提取牙龈卟啉单胞菌ATCC33277株脂多糖(Pg-LPS)。采用ELISA试剂盒定量检测Pg-LPS作用的THP-1和HL-60细胞分泌IL-1β水平。采用TLR2或TLR4抗体阻断试验联合ELISA检测,了解Pg-LPS结合不同靶细胞上Toll样受体的类型。实验中采用大肠杆菌脂多糖(E-LPS)作为Pg-LPS对照。结果1μg/mlPg-LPS作用6h或1μg/mlE-LPS作用24h,THP-1细胞分泌的IL-1β水平明显升高(P<0.01),但Pg-LPS和E-LPS诱生的细胞因子最高浓度相近(P>0.05)。而1μg/mlPg-LPS作用24h或1μg/mlE-LPS分别作用48h,HL-60细胞分泌的IL-1β水平明显升高(P<0.01),且Pg-LPS诱生的最高浓度明显高于E-LPS(P<0.05),但HL-60细胞分泌的上述胞因子最高浓度均明显低于THP-1细胞(P<0.01);TLR2和TLR4抗体均可有效阻断Pg-LPS诱导THP-1细胞分泌IL-1β的活性(P<0.05),但仅发现TLR2抗体可阻断Pg-LPS诱导HL-60细胞分泌IL-1β的活性(P<0.05)。E-LPS诱导THP-1细胞或HL-60细胞分泌IL-1β的活性仅可被TLR4抗体所阻断(P<0.05)。结论Pg-LPS诱导THP-1和HL-60细胞分泌IL-1β高于E-LPS。TLR2和/或TLR4作为Pg-LPS受体取决于细胞种类。Objective To explore the effects of Porphyromonas gingivalis lipopolysaccharide （Pg - LPS） on induction of interleukin -1β （ IL -1β） in THP - 1 and HL - 60 cell and the associated Toll - like receptors. Methods Pg - LPS （ Pg strain, ATCC 33277） was prepared using phenol - water method. The level of IL -1β secreted by THP - 1 cells or HL - 60 cells were quantitatively detected using commercial enzyme - linked immunosorhent assay kit （ELISA）. The blocking test using antibodies against Toll -like receptor （TLR） 2 or TLR4 antibody plus the ELISA were used to determine the types of Pg - LPS binding TLRs on the surface of target cells. A commercial E. coli LPS （ E - LPS） was used as a Pg - LPS control in this study. Results When 1 μg/ml Pg - LPS acted for 6h or 1 μg/ml E - LPS acted for 24h, the levels of IL -1β secreted by THP - 1 cells were obviously increased （ P 〈 0.01 ）. The maximal concentrations of eytokines induced by Pg - LPS were similar to that induced by E - LPS （ P 〉 0.05 ）. When 1 μg/ml Pg - LPS acted for 24h or 1 μg/ml E- LPS acted for 48h, the levels of IL-1β secreted by HL- 60 cells were remarkably increased （ P 〈 0.01 ）. The maximal concentrations of the eytokines secreted by HL - 60 cells were signifieandy lower than that secreted by THP - 1 cells （ P 〈0.01）. The maximal concentrations of IL-1β was obviously higher than that induced by E -LPS （ P 〈 0.05 ）. Anti - TLR2 antibody or anti - TLR4 antibody could block the effects of Pg - LPS inducing THP - 1 cells to secrete IL -1β （ P 〈 0.05 ）, whereas only Anti - TLR2 antibody displayed the inhibition of IL -1β secretion in HL - 60 cells （ P 〈 0.05 ）. Conclusion Pg - LPS shows higher activity inducing THP - 1 or HL - 60 cells to secrete IL -1β compared to E - LPS. TLR2 and/or TLR4 acted as the Pg - LPS receptors is dependent on cell types.

关 键 词：牙龈卟啉单胞菌 脂多糖 细胞 IL-1Β Toll样受体 

分 类 号：R781.4[医药卫生—口腔医学]