细菌细胞壁人髓系细胞触发受体-1天然配体的研究  

Study on the natural ligand（s） of triggering receptor expressed on myeloid cell-1 on bacteria cell wall

作　　者：余志辉 黄红川[1] 毛璞[1] 刘冬冬[1] 莫红缨[1] 何为群[1] 刘晓青[1] 黎毅敏[1] 

机构地区：[1]广州医学院第一附属医院,广州呼吸疾病研究所,广东510120

出　　处：《中国危重病急救医学》2010年第6期335-339,共5页Chinese Critical Care Medicine

基　　金：基金项目：国家自然科学基金资助项目（30871135）

摘　　要：目的 寻找人髓系细胞触发受体-1（TREM-1）的天然配体,为阐明脓毒症的发病机制提供理论依据.方法 分离人外周血中性粒细胞和单核细胞进行原代培养.在两种分离培养的细胞中分别加入热失活的金黄色葡萄球菌、铜绿假单胞菌、结核分枝杆菌或高渗诱导金黄色葡萄球菌L型、铜绿假单胞菌L型处理24 h.在两种分离培养的细胞中分别加入超声提取的金黄色葡萄球菌细胞壁、铜绿假单胞菌细胞壁或结核分枝杆菌细胞壁处理24 h.在两种分离培养的细胞中分别加入各细胞壁三大组分（细胞壁多糖、细胞壁脂类、细胞壁蛋白）处理24 h.用荧光定量聚合酶链反应（PCR）检测各处理组细胞TREM-1 mRNA水平,用酶联免疫吸附法（ELISA）检测各处理组细胞培养上清液中肿瘤坏死因子-α（TNF-α）和白细胞介素-1β（IL-1β）浓度.结果 金黄色葡萄球菌和铜绿假单胞菌的菌体、细胞壁和细胞壁多糖处理后中性粒细胞和单核细胞TREM-1mRNA水平以及细胞培养上清液中TNF-α和IL-1β浓度均明显升高.与空白对照组相比,金黄色葡萄球菌细胞壁多糖处理后中性粒细胞TREM-1 mRNA水平增加了（3.86±0.20）倍,单核细胞TREM-1 mRNA水平增加了（5.15±0.56）倍（均P＜0.05）;铜绿假单胞菌细胞壁多糖处理后中性粒细胞TREM-1 mRNA水平增加了（4.03±0.15）倍,单核细胞TREM-1 mRNA水平增加了（7.22±0.73）倍（均P＜0.05）.加入能竞争性结合TREM-1配体的LP17可以削弱此效应;而结核分枝杆菌的菌体、细胞壁和细胞壁组分均无此效应.结论 人TREM-1天然配体位于金黄色葡萄球菌和铜绿假单胞菌的细胞壁上,成分可能为细菌细胞壁多糖.：Objective To look for the natural ligand（s） of human triggering receptor expressed on myeloid cell-1 （TREM-1）, in order to provide the theoretical basis for elucidation of the pathogenesis of sepsis.Methods Neutrophils and monocytes isolated from human peripheral blood were treated with heat-inactivated Staphylococcus aureus , Pseudomonas aeruginosa , Mycobacterium tuberculosis, Staphylococcus aureus L-form or Pseudomonas aeruginosa L-form respectively for 24 hours.The cell wall was extracted from Staphylococcus aureus, Pseudomonas aeruginosa and Mycobacterium tuberculosis by ultrasound.Neutrophils and monocytes were isolated and treated with the cell wall respectively for 24 hours.Neutrophils and monocytes were isolated and treated with three main components from bacterial cell wall （polysaccharides,lipids and proteins） respectively for 24 hours.The level of TREM-1 mRNA was measured with fluorescent quantitative polymerase chain reaction （PCR）, and the concentrations of tumor necrosis factor-α （TNF-α）and interleukin-1β （IL-1β） were measured with enzyme-linked immunosorbent assay （ELISA）.Results The TREM-1 mRNA level and the concentrations of TNF-α and IL-1β in cell supernatant of neutrophils and monocytes were upgraded when treated with cell, cell wall and cell wall polysaccharides of Staphylococcus aureus and Pseudomonas aeruginosa.Compared with the blank control group, the TREM-1 mRNA level of neutrophils and monocytes was upgraded to （3.86± 0.20）-fold and （5.15±0.56）-fold respectively when treated with cell wall polysaccharides of Staphylococcus aureus （both P〈0.05）;the TREM-1 mRNA level of neutrophils and monocytes was upgraded to （4.03±0.15）-fold and （7.22±0.73）-fold respectively when treated with cell wall polysaccharides of Pseudomonas aeruginosa （both P〈0.05）.The effect could be attenuated by the addition of LP17 which could bind TREM-1 ligand.This attenuating effect was not found when the cells were treated with cell, cell wall or cell

关 键 词：脓毒症 髓系细胞触发受体-1 配体 多糖 

分 类 号：R459.7[医药卫生—急诊医学]