内毒素耐受对人中性粒细胞抗炎反应的影响
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国家自然科学基金(81370025);江苏省自然科学基金(BK20161565)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项目(2014-37)


Effects of endotoxin tolerance on anti-inflammatory responses in human neutrophils
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    摘要:

    目的:探讨脂多糖(lipopolysaccharide,LPS)诱导的耐受对人中性粒细胞分泌抗炎因子白介素(interleukin,IL)-10及细胞凋亡水平的影响。方法:采用1 μg/mL牙龈卟啉单胞菌(Porphyromonasgingivalis,P.gingivalis)LPS或者1 μg/mL大肠杆菌(Escherichia coli,E.coli)LPS刺激人中性粒细胞12 h,诱导细胞耐受。洗脱后,分别采用2种LPS再次刺激20 h,采用ELISA技术检测细胞分泌IL-10水平。同法诱导细胞耐受,洗脱后,再次刺激3 h,采用AV-PI法检测细胞凋亡水平。结果:P.gingivalis LPS或E.coli LPS单次刺激人中性粒细胞后,IL-10分泌水平均较刺激前明显增加(P<0.05),凋亡水平则无明显改变(P>0.05)。2种LPS分别重复刺激之后,IL-10分泌水平均较单次刺激后显著增加(P<0.05)。P.gingivalis LPS重复刺激3 h后,中性粒细胞短期内凋亡水平与单次刺激后无明显改变(P>0.05),而E.coli LPS重复刺激3 h后,细胞短期内凋亡水平较单次刺激后明显下降(P<0.05)。结论:内毒素耐受能促进人中性粒细胞分泌IL-10,但不同LPS诱导的耐受对细胞凋亡水平的影响可能存在差异。

    Abstract:

    Objective: To observe the effects of endotoxin tolerance induced by lipopolysaccharide (LPS) on the production of anti-inflammatory cytokine IL-10 and the level of apoptosis in human neutrophils. Methods: Neutrophils were pretreated with 1 μg/mL Porphyromonasgingivalis (P.gingivalis) LPS or 1 μg/mL Escherichia coli (E.coli) LPS for 12 h to induce cell tolerance. Then, the cells were washed and stimulated with the same LPS for additional 20 h (for ELISA) or 3 h (for flow cytometry). Levels of IL-10 in supernatants and apoptosis of neutrophils were detected by ELISA and flow cytometry, respectively. Results: After 20 h, the amount of IL-10 secreted by neutrophils treated with P.gingivalis LPS or E.coli LPS was increased significantly compared with that without any stimulation (P<0.05), while no significant changes of apoptosis in neutrophils with LPS stimulation were observed after 3 h (P>0.05). After restimulation with P.gingivalis LPS or E.coli LPS, IL-10 production was increased significantly compared with that secreted by neutrophils challenged with the same LPS only once (P<0.05). However, no significant changes of apoptosis in the short term were detected after P.gingivalis LPS retreatment for 3 h (P>0.05), and apoptosis levels in the short term were significantly decreased after E.coli LPS restimulations for 3 h (P<0.05). Conclusion: Endotoxin tolerance might contribute to the increased production of IL-10 in human neutrophils, but the effects of tolerance on apoptosis might depend on the types of LPS.

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陈 杨,祝祥清,程筱番,仇嘉颖,徐 艳,孙 颖.内毒素耐受对人中性粒细胞抗炎反应的影响[J].南京医科大学学报(自然科学版),2017,(1):30-34

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  • 收稿日期:2016-04-28
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  • 在线发布日期: 2017-02-16
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