南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (06): 842-.

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甘露聚糖结合凝集素对人中性粒细胞功能的影响

陈阿德,王燕,张丽芸,卢晓,左大明,陈政良   

  • 出版日期:2013-06-20 发布日期:2013-06-20

Effects of mannan-binding lectin on the functions of human polymorphonuclear cells

  • Online:2013-06-20 Published:2013-06-20

摘要: 目的研究甘露聚糖结合凝集素(MBL)对人中性粒细胞(PMN)功能的影响。方法采用ELISA和Dot blot检测MBL与微
生物的结合;应用流式细胞术检测及荧光显微镜观察MBL对PMN吞噬功能的影响;用qPCR分析MBL对PMN表达IL-1β、
TNF-α和CD11b mRNA的影响,以ELISA检测MBL对PMN分泌TNF-α和IL-6的影响;应用NBT还原法检测MBL对PMN呼
吸爆发的影响。结果MBL与微生物结合并呈浓度依赖关系。无人血清时,MBL对PMN吞噬C. albicans 和E.coli 无明显影
响。存在MBL缺陷人混合血清时,MBL可促进PMN对C. albicans的吞噬且这种作用可被甘露聚糖所阻断,上调PMN IL-1β、
TNF-α、IL-6及CD11b表达水平,刺激PMN呼吸爆发。结论MBL以补体凝集素途径依赖方式增强PMN吞噬功能,并促进炎性
细胞因子分泌。

Abstract: Objective To investigate the effects of mannan-binding lectin (MBL) on the functions of human polymorphonuclear
cells (PMNs). Methods ELISA and Dot blot were performed to examine the binding between MBL and the microorganisms.
Flow cytometry and fluorescence microscopy were employed to analyze the phagocytosis of FITC-labeled microognisms by
the PMNs. Real-time quantitative PCR was used to detect the expression levels of IL-1β, TNF-α and CD11b mRNA in the
PMNs, and ELISA used to detect the levels of TNF-α and IL-6 in the supernatants of PMN culture. Nitro-blue tetrazolium
reduction assay was used to estimate the levels of superoxide production. Results MBL bound to the microorganisms in a
dose-dependent manner. MBL had no significant effect on phagocytosis of C. albicans and E.coli by the PMNs in the absence of
human serum, but in presence of mixed MBL-deficient human sera, MBL promoted the phagocytosis of C. albicans, which
could be blocked by mannan. Mannan treatment increased the expressions of IL-1β, TNF-α, IL-6 and CD11b and enhanced
superoxide production in the PMNs. Conclusion MBL can promote phagocytosis of microorganisms by PMNs and increase
the expressions of proinflamatory cytokines from PMNs in a complement lectin pathway-dependent manner.