文献类型：期刊文献

中文题名：人单核-巨噬细胞和中性粒细胞杀灭问号钩端螺旋体能力差异及其机制

英文题名：Human mononuclear macrophages and neutrophils： differences in killing ability and mechanism against Leptospira interrogans

作者：阮萍[1];林旭瑷[2];张颖颖[2];周海丽[2];严杰[2];陈旭[2]

机构：[1]绍兴文理学院医学院医学检验系,312000;[2]浙江大学医学院病原生物学系,杭州310058

年份：2017

卷号：37

期号：6

起止页码：412

中文期刊名：中华微生物学和免疫学杂志

外文期刊名：Chinese Journal of Microbiology and Immunology

收录：CSTPCD、、北大核心2014、Scopus、CSCD2017_2018、北大核心、CSCD

基金：国家自然科学基金（81671974,81471907）

语种：中文

中文关键词：问号钩端螺旋体;单核-巨噬细胞;中性粒细胞;活性氧;一氧化氮;胞内游离Ca2+;Intracellular;free;Ca2+

外文关键词：Leptospira interrogans;Mononuclear macrophage;Neutrophil;Reactive oxygen species;Nitric oxide

中文摘要：目的 了解人单核-巨噬细胞和中性粒细胞杀灭问号钩端螺旋体（简称钩体）能力差异及其机制.方法 人THP-1单核细胞和HL-60细胞分别用佛波酯（PMA）和全反式维甲酸（ATRA）预处理,使其分化为单核-巨噬细胞和中性粒细胞.采用激光共聚焦显微镜法检测问号钩体黄疸出血群赖型赖株感染THP-1单核-巨噬细胞和HL-60中性粒细胞内总活性氧（ROS）、一氧化氮（NO）水平和游离钙离子浓度（[Ca2＋]i）变化.采用荧光分光光度法检测总ROS、NO抑制剂和胞内游离Ca2＋螯合剂（NAC、SMT和BAPTA/AM）作用前后THP-1单核-巨噬细胞和HL-60中性粒细胞杀灭胞内问号钩体的能力及其差异.结果 THP-1单核-巨噬细胞和HL-60中性粒细胞感染问号钩体后总ROS、NO水平和[Ca2＋]i均显著升高（P〈0.05）,但前者总ROS、NO水平和[Ca2＋]i显著高于后者（P〈0.05）.THP-1单核-巨噬细胞杀灭胞内问号钩体能力显著强于HL-60中性粒细胞（P〈0.05）.抑制胞内总ROS、NO和游离Ca2＋可导致THP-1单核-巨噬细胞杀灭胞内问号钩体能力显著下降（P〈0.05）,但对HL-60中性粒细胞无明显影响.结论 单核-巨噬细胞而非中性粒细胞是感染时清除问号钩体的主要吞噬细胞,高水平总ROS、NO和胞内游离Ca2＋与单核-巨噬细胞杀灭问号钩体能力密切相关.

外文摘要：Objective To understand the differences in killing ability and mechanism of human mononuclear macrophages and neutrophils against Leptospira interrogans.Methods Human THP-1 and HL-60 cell lines were respectively pretreated with PMA （phorbol 12-myristate 13-acetate） and ATRA （all-trans retinoic acid） to induce their differentiation into macrophages and neutrophils.Confocal microscopy was used to detect the changes in total ROS （reactive oxygen species） and NO （nitric oxide） levels as well as free Ca2＋ concentration （[Ca2＋]i） in THP-1 monocyte-derived macrophages and HL-60 cell-derived neutrophils after infection with Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai.Fluorospectrophotometry was applied to analyze the differences in killing ability between THP-1 monocyte-derived macrophages and HL-60 cell-derived neutrophils against intracellular leptospires before and after treatment with total ROS and NO inhibitors and intracellular free Ca2＋ chelator.Results The total ROS and NO levels and [Ca2＋]i in THP-1 monocyte-derived macrophages and HL-60 cell-derived neutrophils were significantly increased after infection with the spirochete （P〈0.05）.Moreover,the total ROS and NO levels and [Ca2＋]i in the former were significantly higher than those in the latter （P〈0.05）.The THP-1 monocyte-derived macrophages had stronger killing ability against intracellular leptospires than the HL-60 cell-derived neutrophils （P〈0.05）.Inhibiting total intracellular ROS,NO or free Ca2＋ could result in decreased killing ability of THP-1 monocyte-derived macrophages against intracellular leptospires,but not affect the killing ability of HL-60 cell-derived neutrophils.Conclusion Mononuclear macrophages rather than neutrophils act as the main phagocytes eliminating Leptospira interrogans.High levels of total intracellular ROS,NO and free Ca2＋ are closely associated with the ability of mononuclear macrophages to kill Leptospira interrogans.