白细胞介素-10对小鼠中性粒细胞吞噬和杀菌活性的影响
Effects of interleukin-10 on phagocytosis and bactericidal activity of neutrophils in mouse model
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摘要: 目的: 探讨白细胞介素-10(IL-10)对中性粒细胞(PMN)吞噬和杀菌活性的影响,以了解IL-10对非特异性免疫的作用机制。方法: 小鼠麻醉后从眼球取血,与不同浓度IL-10在37℃水浴中作用2 h,然后加FITC标记大肠埃希菌,置37℃水浴中15 min,溶血剂去除红细胞,流式细胞仪检测各IL-10浓度组PMN的吞噬率。小鼠麻醉后局部消毒,摘眼球,无菌方法取血。与不同浓度IL-10在37℃下作用2 h,然后加大肠埃希菌置37℃水浴1 h,同时设PMN杀菌0 min组、无IL-10的PMN杀菌对照组,各组样品作适当稀释后接种琼脂平板,最后观察各组菌落数,与对照组比较,计算IL-10对杀菌力影响。结果: PMN的吞噬功能在IL-10浓度为1 ng/ml组减低(15.87 ±14.25)%,但差异无统计学意义(P > 0.05)。而PMN的杀菌功能在IL-10浓度为0.1 ng/ml组降低(39.59 ±37.36)%,在IL-10浓度为1 ng/ml组则降低(42.28 ±22.24)%,差异均有统计学意义(P <0.05)。结论: IL-10对PMN的吞噬功能有轻度的抑制作用,对PMN的杀菌功能有明显的抑制作用,对非特异性免疫亦有抑制作用。Abstract: Objective: To investigate the effects of inteluekin-10(IL-10) on the neutrophil phagocytosis and bactericidal activity in mouse model. Methods: Whole blood of mice was pretreated with IL-10 at different concentrations,ranged from 1 pg to 100 ng/ml for 2 hours at 37℃,and then incubated with FITC labeled E. coli at 37℃ for 15 minutes. After lysing red blood cells the neutrophil phagocytosis was measured by flow cytometry. In the bactericidal assay,whole blood of mice pretreated with IL-10(0.1 and 1 ng/ml) for 2 hours at 37℃ was incubated with E. coli in water bath at 37℃ for 1 hour,and then cultured on agar plates. The number of colonies was counted and the inhibitory effect of IL-10 on killing activity was calculated. Results: The reduced rates of phagocytosis was (15.87 ±14.25)% only in the group that pretreated with IL-10 at 1 ng/ml,but with no significance(P > 0.05). The relative rates of bactericidal activity of neutrophil in the groups of IL-10 at 0.1 ng/ml and 1 ng/ml were significanty reduced(39.59 ±37.36)% and (42.28 ±22.24)% (P < 0.05),respectively. Conclusions: IL-10 slightly decreases the phagocytosis of the neutrophils,but potently suppresses the bactericidal activity of neutrophils. IL-10 is capabling of down-regulating non-specific immunity.