龈下菌斑诱导的耐受对人巨噬细胞分泌TNF-α和IL-10的影响及机制
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国家自然科学基金(81370025);江苏省高等学校大学生实践创新训练计划(201310312017Z);江苏高校优势学科建设工程资助(2014-37)


Effects and mechanisms of endotoxin tolerance induced by subgingival plaques on the productions of TNF-α and IL-10 in human macrophages
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    摘要:

    目的:观察龈下菌斑诱导的耐受对人巨噬细胞分泌炎症因子肿瘤坏死因子α(tumor necrosis factor α,TNF-α)和抗炎因子白介素-10(inter leukin 10,IL-10)的影响,以及与Toll样受体(Toll-like receptor,TLR)2-TLR4的关系。方法:采集中-重度慢性牙周炎患者及健康人的龈下菌斑,分别重复刺激,诱导人巨噬细胞产生耐受。采用ELISA 技术检测TNF-α和IL-10表达水平的变化,采用流式细胞技术检测TLR2-TLR4表达水平的改变。结果:2种菌斑第1次刺激巨噬细胞后,TNF-α和IL-10水平均较刺激前明显增高(P < 0.05)。2种菌斑重复刺激后,TNF-α分泌水平均较第1次刺激后明显降低(P < 0.05),IL-10水平均与第1次刺激后无明显差别(P > 0.05)。尽管2种菌斑第1次刺激后,TLR2-TLR4表达水平均较刺激前明显升高(P < 0.05),2种菌斑重复刺激后,TLR2-TLR4表达水平均与第1次刺激后无明显差别(P > 0.05)。结论:龈下菌斑诱导的耐受能够抑制TNF-α的分泌,进而可能影响牙周组织的炎症和免疫反应,但这种改变与TLR2-TLR4表达水平无关。

    Abstract:

    Objective:To study the effects of endotoxin tolerance induced by subgingival plaques on the productions of inflammatory cytokine TNF-α and anti-inflammatory cytokine IL-10 and the protein expressions of Toll-like receptor 2, 4 (TLR2, 4)in human macrophages. Methods:Macrophages were pretreated with subgingival plaques from moderate to serve chronic periodontitis patients or healthy controls (24h), washed (2h)and treated with the same plaques again (24 h). Levels of TNF-α and IL-10 in supernatant were detected by ELISA. Moreover, TLR2, 4 protein expressions in these cells were explored by flow cytometry. Results:The amounts of TNF-α and IL-10 secreted by macrophages stimulated with both kinds of subgingival plaques were increased significantly after 24h(P < 0.05). After repeated stimulation with the same plaques, secretions of TNF-α were decreased significantly compared with those following single challenge(P < 0.05), while only comparable levels of IL-10 were confirmed in both two kinds of plaques- tolerized cells(P > 0.05). However, no significant changes of TLR2 and 4 were detected in macrophages retreated with these plaques compared with those following single stimulations. Conclusion:Secretions of TNF-α could be suppressed by endotoxin tolerance induced by subgingival plaques, which might contribute to limiting periodontal inflammation. However, development of this tolerance might not be affected by TLR2 and 4.

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卢 伟,贡丹军,孙梦君,徐 艳,孙 颖.龈下菌斑诱导的耐受对人巨噬细胞分泌TNF-α和IL-10的影响及机制[J].南京医科大学学报(自然科学版),2015,(5):722-726

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  • 收稿日期:2015-01-13
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  • 在线发布日期: 2015-05-22
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