内毒素耐受对THP-1细胞分泌TNF-α和IL-10的影响

Effects of endotoxin tolerance on the production of TNF-α and IL-10 in THP-1 Cells

目的:观察细菌脂多糖(lipopolysaccharides,LPS)反复刺激人单核细胞株THP-1细胞,诱导产生的内毒素耐受对该细胞分泌炎症因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和抗炎因子白细胞介素(interleukin,IL)-10的影响。方法:采用1μg/ml牙龈卟啉单胞菌(Porphyromonas gingivalis,P.gingivalis)LPS或1μg/ml大肠杆菌(Escherichia coli,E.coli)LPS刺激THP-1细胞24 h,洗脱后,分别采用相同LPS再刺激24 h,构建内毒素耐受模型。采用ELISA技术检测细胞条件培养液中TNF-α和IL-10分泌水平的变化。结果:P.gingivalis LPS或E.coli LPS刺激THP-1细胞24 h后,TNF-α和IL-10分泌水平均较刺激前明显增高(P<0.05)。2种LPS重复刺激后,TNF-α分泌水平较第1次刺激后明显降低(P<0.05),IL-10分泌水平则较第1次刺激后明显增高(P<0.05)。结论:内毒素耐受能抑制THP-1细胞分泌TNF-α,促进该细胞分泌IL-10,进而可能影响牙周组织的炎症和免疫反应。

Objective：To observe the effects of endotoxin tolerance on the production of inflammatory cytokine TNF-α and antiinflammatory cytokine IL-10 in THP-1 cells.Methods：THP-1 cells were pretreated with 1 μg / ml Porphyromonas gingivalis（P. gingivalis） lipopolysaccharides（LPS） or 1μg / ml Escherichia coli（E.coli） LPS for 24 h.Then,the cells were washed and stimulated with the same LPS for additional 24 h.The levels of TNF-α and IL-10 in supernatants were detected by ELISA.Results：The amounts of TNF-α and IL-10 secreted by THP-1 cells stimulated with P.gingivalis LPS or E.coli LPS were increased significantly after 24 h（P 0.05）.After restimulations for another 24 h,TNF-α production was decreased significantly compared with that secreted by the cells stimulated with the same LPS only once（P 0.05）,while the level of IL-10 was increased significantly（P 0.05）.Conclusion： Repeated LPS stimulations triggered endotoxin tolerance and led to a decrease in TNF-α production and an increase in IL-10 production,which might have effects on periodontal inflammation.