内毒素诱导正常人单核细胞活化中CD69、TNF-α、IL-6的表达变化及意义  

The change and significance of CD69, TNF-α, IL-6 in the monocytes stimulated by endotoxin of healthy people

作　　者：李珊珊[1] 李璐[1] 张丹丹[1] 冯霞[1] 丁美[1] 陈煜[1] 

机构地区：[1]首都医科大学附属北京佑安医院人工肝中心,100069

出　　处：《北京医学》2013年第12期1003-1006,I0002,共5页Beijing Medical Journal

基　　金："十二五"国家科技重大专项(2012ZX 1000-2004-006);北京市卫生系统高层次卫生技术人才培养计划(2013-3-071);北京市科技计划课题(Z131100004613030)

摘　　要：目的探讨内毒素诱导正常人单核细胞活化时抗原分子CD69、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的变化及意义。方法采集10例健康人外周静脉血20 ml,经乙二胺四乙酸(EDTA)抗凝。通过Ficoll-Hypaque联合Percoll分离液分离纯化单核细胞后体外培养,设未干预组为对照,实验组予内毒素分别刺激30 min、1、2、4 h后收集细胞与上清液。通过流式细胞术检测单核细胞表面CD69的阳性细胞百分比,Luminex方法检测上清液中TNF-α、IL-6的浓度。结果分离的单核细胞纯度达95.5%。单核细胞表面CD69的表达率在内毒素刺激2、4 h分别为(25.1±1.8)%、(41.7±1.4)%,明显高于对照组[(22.1±1.4)%,P均﹤0.05]。上清液中TNF-α在内毒素刺激2、4 h分别为(453.654 6±163.616 8)pg/ml、(1 633.787 8±528.839 9)pg/ml,明显高于对照组[(0.119 1±0.213 5)pg/ml,P均﹤0.05]。内毒素刺激4 h时IL-6为(1464.451 0±488.378 3)pg/ml,明显高于对照组[(0.053 9±0.095 5)pg/ml,P﹤0.05]。结论CD69、TNF-α、IL-6的表达上调与内毒素诱导单核细胞的活化密切相关。Objective To investigate the change and expression of CD69,TNF-α,IL-6 in monocytes isolated from healthy people stimulated by endotoxin in vitro. Methods Twenty milliliter peripheral blood was sampled and anticoagulant from 10 healthy people. Monocytes were separated from the blood using Ficoll lymphocyte and Percoll isolation, then the purity of monocytes were checked. The untreated cells served as control group while the experimental group was stimulated by endotoxin （100 ng/ml）. After cultured for 30 rain, l,2,4 h, cells and supernate were collected, CD69 expression was detceted by flow eytometry, TNF-α and IL-6 level was examined by Luminex. Results Monocytes purity reached 95.5% by using Fieoll and Percoll isolation. After stimulation by endotoxin, CD69 expression in the stimulant group at 2,4 h[（25.1±1.8）%,（41.7±1.4）%] was significantly higher than that in the control group [（22.1±1.4）%, P〈 0.05]. The expression of TNF-α which secreted from monocytes became significantly higher in the stimulant group than that in the control group at 2,4 h [（453,654 6±163.616 8）pg/ml, （1 633.787 8±528.839 9）pg/ml vs. （0.119 1±0.213 5）pg/ml, P 〈 0.05]. IL-6 was significantly higher in the stimulant group than that in the control group at 4h [（1 464.451 0±488.378 3） pg/ml vs. （0.053 9±0.095 5）pg/ml, P〈 0.05]. Conclusion The upexpression of CD69, TNF-α, IL-6 is associated with the activation of the monocytes stimulated by endotoxin.

关 键 词：内毒素 单核细胞 CD69 肿瘤坏死因子-α 白细胞介素-6 

分 类 号：R575.3[医药卫生—消化系统]