›› 2018, Vol. 38 ›› Issue (3): 193-196.

• 基础研究 •    下一篇

内毒素耐受对共培养的中性粒细胞和单核细胞分泌TNF-α和IL-8的影响

方媛春1,2,祝祥清1,2,顾建雨1,2,尹兰兰1,2,孙颖3   

  1. 1.
    2. 南京医科大学附属口腔医院 牙周科
    3. 南京医科大学附属口腔医院牙周科
  • 收稿日期:2017-09-13 修回日期:2017-10-28 出版日期:2018-03-28 发布日期:2018-03-16
  • 通讯作者: 孙颖 E-mail:ebolasun@njmu.edu.cn
  • 基金资助:
    国家自然科学基金;国家自然科学基金;江苏高校优势学科建设工程资助项目

Influence of endotoxin tolerance on TNF-α and IL-8 production in co-cultures of neutrophils and THP-1 cells

  • Received:2017-09-13 Revised:2017-10-28 Online:2018-03-28 Published:2018-03-16

摘要: 目的 观察脂多糖(lipopolysaccharides,LPS)诱导的耐受对共培养的中性粒细胞和人单核细胞株THP-1细胞表达抗炎因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和趋化因子白介素-8(Inter leukin-8, IL-8)水平的影响。 方法 采用1 μg/mL牙龈卟啉单胞菌(Porphyromonas gingivalis, P. gingivalis)LPS 或1 μg /mL大肠杆菌(Escherichia coli,E.coli)LPS分别刺激共培养的中性粒细胞和THP-1细胞24 h,洗脱后,再次刺激24 h,诱导细胞产生耐受。采用ELISA技术检测细胞条件培养液中 TNF-α 和 IL-8 表达水平的变化。 结果 P.gingivalis LPS或E.coli LPS单次刺激后,共培养的中性粒细胞和THP-1细胞分泌TNF-α和IL-8的水平均较刺激前明显增高(P<0.05);P.gingivalis LPS和E.coli LPS重复刺激后,共培养的中性粒细胞和THP-1细胞分泌的TNF-α水平较单次刺激后明显降低(P<0.05),但IL-8水平较单次刺激后明显增高(P<0.05)。 结论 共培养的中性粒细胞和单核细胞产生的内毒素耐受可能抑制TNF-α表达,促进IL-8表达,进而可能影响牙周组织的炎症和免疫反应。

关键词: 内毒素耐受, 共培养, 中性粒细胞, THP-1细胞

Abstract: Objective To observe the effects of endotoxin tolerance triggered by repeated stimulation of lipopolysaccharide (LPS) on TNF-α and IL-8 expressions in co-cultures of neutrophils and THP-1 cells. Methods Co-cultures of neutrophils and THP-1cells were pretreated with 1 μg/mL Porphyromonas gingivalis(P.gingivalis) LPS or 1 μg/mL Escherichia coli(E.coli) LPS for 24 h, washed and stimulated by the same LPS again for another 24 h. The levels of TNF-α and IL-8 in supernatants were detected by ELISA. Results The expression levels of TNF-α and IL-8 significantly increased in co-cultured cells stimulated by P.gingivalis LPS or E.coli LPS compared with those without any stimulation (P<0.05). The expression levels of TNF-α in cells restimulated by P.gingivalis LPS or E.coli LPS were obviously down-regulated compared with those stimulated by the same LPS only once (P<0.05), while the expression levels of IL-8 were significantly up-regulated in comparison with those secreted by co-cultures of neutrophils and THP-1 cells stimulated by the same LPS only once (P<0.05). Conclusion Endotoxin tolerance may suppress the secretion of TNF-α and promote the production of IL-8 in co-cultures of neutrophils and THP-1 cells.

Key words: endotoxin tolerance, co-culture, neutrophils, THP-1 cells

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