内毒素刺激诱导人外周血细胞促炎细胞因子的释放

Effects of lipopolysaccharide stimulation on secretion of pro-inflammatory cytokines from mononuclear cells of human peripheral blood

目的：探讨内毒素（LPS）体外刺激对人外周血细胞高迁移率族蛋白B1（HMGB1）及肿瘤坏死因子（TNF）-α、白介素（IL）-6和IL-10分泌的影响.方法：采集20例健康志愿者外周血,与培养基等体积混合后体外培养,给予不同浓度LPS（100 ng/ml,500 ng/ml）刺激,分别于刺激后0、2、6、12、24 h收集细胞培养上清液,ELISA法检测其HMGB1和TNF-α、IL-6、IL-10的含量.结果：体外LPS刺激诱导外周血细胞分泌HMGB1增加,12 h后表达持续升高（P〈0.01）,分泌水平呈明显时间依赖性;TNF-α、IL-6产生亦呈时间、剂量依赖性,分别于24 h和6～12 h达分泌高峰（P〈0.01）;IL-10表达量均较低.结论：TNF-α、IL-6为早期炎症介质,HMGB1为介导内毒素所致迟发死亡的重要晚期炎症介质,它们在脓毒症的发生、发展中可能扮演重要角色.

Objective：To observe the effect of lipopolysaccharide （LPS） stimulation on secretion of high mobility group box-1 protein （HMGB1）, tumor necrosis factor-α （TNF-α）, interleukin 6 （IL-6） and IL-10 from cells of human peripheral blood in vitro. Methods： Peripheral blood samples were collected from 20 healthy volunteers and cultured with the equal volume of cell culture medium in vitro. After treatment with LPS （100 ng/ml or 500 ng/ml） for 0, 2, 6, 12, or 24 hours, culture supernatants were collected and levels of HMGB1, TNF α, IL-6 and IL-10 were measured with ELISA. Results： A significant release of HMGB1 in human blood was observed after cultured with LPS for 12 hours （P（0. 01）, showing a time-dependent response manner. Similarly, production of TNF-α and IL-6 peaked at 24 hours and 6-12 hours in a dose-dependent manner after stimulation with LPS （P〈0. 01）. Expression of IL-10 remained in low level after stim- ulation with LPS in various doses. Conclusions： HMGB1 is a late inflammatory factor in inducing the late death as a result of endotoxin challenge. TNF-α and IL-6 are important early pro-inflammatory cytokines acting as the motivator in the de velopment of sepsis. Both early and late pro-inflammatory cytokines appear to be involved in the development of sepsis.