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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1μg/m L牙龈卟啉单胞菌(Porphyromonasgingivalis,P.gingivalis)LPS或者1μg/m L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。
Objective: To investigate the effects of lipopolysaccharide (LPS) -induced tolerance on the secretion of anti-inflammatory cytokines IL-10 and apoptosis in human neutrophils. Methods: Human neutrophils were stimulated with 1 μg / mL LPS or Porphyromonas gingivalis (LPS) or 1 μg / mL Escherichia coli (E. coli) LPS for 12 h. . After elution, the cells were stimulated with two kinds of LPS again for 20 h, and the levels of IL-10 secreted by the cells were measured by ELISA. The same method of induction of cell tolerance, after elution, re-stimulation 3 h, AV-PI detection of apoptosis levels. Results: After single stimulation of human neutrophil by P. gingivalis LPS or E. coli LPS, the level of IL-10 secretion increased significantly (P <0.05) and the level of apoptosis did not change significantly (P> 0.05) . After repeated stimulation with LPS, the level of IL-10 secretion increased significantly (P <0.05) after single stimulation. After repeated stimulation of P.gingivalis LPS for 3 h, the apoptosis of neutrophils in the short term did not change significantly after single stimulation (P> 0.05), while the apoptosis of cells in 3 h after repeated stimulation by E. coli LPS After a single stimulus decreased significantly (P <0.05). Conclusion: Endotoxin tolerance can promote the secretion of IL-10 by human neutrophils, but the effects of LPS-induced tolerance on the level of apoptosis may be different.