Baqui A A, Meiller T F, Chon J J, Turng B F, Falkler W A
Department of Oral Medicine, Dental School, University of Maryland, Baltimore, USA.
Oral Microbiol Immunol. 1998 Jun;13(3):173-80. doi: 10.1111/j.1399-302x.1998.tb00729.x.
This study focused on the effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) and lipopolysaccharide of the putative periodontal pathogens Porphyromonas gingivalis or Fusobacterium nucleatum on IL-6 production by THP-1 cells (a human monocytic cell line). Resting THP-1 cells were alternatively treated with GM-CSF (50 IU/ml) and lipopolysaccharide of P. gingivalis or F. nucleatum, in varying concentrations for varying time periods. IL-6 production in supernatant fluids of treated cells was evaluated by an enzyme-linked immunosorbent assay (ELISA) and a reverse transcription polymerase chain reaction (RT-PCR) was used to evaluate gene expression. Untreated THP-1 cells did not produce IL-6 as determined by ELISA. RT-PCR also failed to detect IL-6 mRNA in untreated THP-1 cells, indicating that IL-6 was not constitutively produced. After stimulation of THP-1 cells with lipopolysaccharide of F. nucleatum or P. gingivalis, IL-6 was produced, peaking at 4 h (200-300 pg/ml) and thereafter sharply declining by 8 h. When GM-CSF was added together with lipopolysaccharide of P. gingivalis or F. nucleatum, there was a synergistic quantitative increase in production of IL-6 as measured by ELISA as compared with lipopolysaccharide alone. IL-6 mRNA was detected by RT-PCR, 15 min after stimulation with lipopolysaccharide of either P. gingivalis or F. nucleatum. GM-CSF supplementation with lipopolysaccharide of P. gingivalis shortened the transcription of IL-6 mRNA to 5 min, a shift which was not observed with lipopolysaccharide of F. nucleatum, possibly indicating a different mechanism of initiation of transcription. Production of IL-6 by GM-CSF-treated THP-1 cells in the presence of lipopolysaccharide of oral microorganisms may provide a model for studying the role of macrophages in acute and chronic periodontal diseases, including the clinical periodontal exacerbation as observed in chemotherapy patients receiving GM-CSF for bone marrow recovery.
本研究聚焦于粒细胞巨噬细胞集落刺激因子（GM-CSF）以及假定的牙周病原体牙龈卟啉单胞菌或具核梭杆菌的脂多糖对THP-1细胞（一种人单核细胞系）产生白细胞介素-6（IL-6）的影响。静息的THP-1细胞分别用GM-CSF（50 IU/ml）以及不同浓度的牙龈卟啉单胞菌或具核梭杆菌脂多糖处理不同时间段。通过酶联免疫吸附测定（ELISA）评估处理后细胞上清液中IL-6的产生情况，并采用逆转录聚合酶链反应（RT-PCR）评估基因表达。ELISA检测显示，未处理的THP-1细胞不产生IL-6。RT-PCR也未能在未处理的THP-1细胞中检测到IL-6 mRNA，表明IL-6不是组成性产生的。用具核梭杆菌或牙龈卟啉单胞菌脂多糖刺激THP-1细胞后，可产生IL-6，在4小时达到峰值（200 - 300 pg/ml），此后到8小时急剧下降。当GM-CSF与牙龈卟啉单胞菌或具核梭杆菌脂多糖一起添加时，与单独使用脂多糖相比，ELISA检测显示IL-6的产生在数量上有协同增加。在用牙龈卟啉单胞菌或具核梭杆菌脂多糖刺激15分钟后，通过RT-PCR检测到IL-6 mRNA。GM-CSF与牙龈卟啉单胞菌脂多糖共同作用可将IL-6 mRNA的转录缩短至5分钟，而具核梭杆菌脂多糖未观察到这种变化，这可能表明转录起始机制不同。在存在口腔微生物脂多糖的情况下，GM-CSF处理的THP-1细胞产生IL-6可能为研究巨噬细胞在急性和慢性牙周疾病中的作用提供一个模型，包括在接受GM-CSF促进骨髓恢复的化疗患者中观察到的临床牙周病情加重。
