Poly(I∶C)和LPS共刺激对人气道上皮细胞趋化因子表达的影响及机制  

Effects on chemotactic factor expression in bronchial epithelial cells by co-stimulation of poly(I:C) and lipopolysaccharide and the underlying mechanism

作　　者：张景熙[1] 胥武剑[1] 韩一平[1] 白冲[1] 李强[1] 

机构地区：[1]上海第二军医大学附属长海医院呼吸内科,上海200433

出　　处：《细胞与分子免疫学杂志》2012年第10期1046-1050,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81100017)

摘　　要：目的:研究模拟病毒感染及内毒素共刺激对气道上皮细胞趋化因子表达的影响。方法:人气道上皮细胞(16HBE)给予聚肌胞苷酸[poly(I∶C)]、内毒素(LPS)、地塞米松及p38MAPK抑制剂(SB203580)处理,用RT-PCR检测刺激6 h后IL-8、干扰素诱导蛋白10(IP-10)mRNA的转录水平,用ELISA检测刺激24 h后培养上清液中IL-8和IP-10蛋白含量。结果:10μg/mL LPS刺激后IL-8 mRNA及蛋白表达较对照组升高,IP-10 mRNA及蛋白表达未见明显升高;加入0.1μg/mL poly(I∶C)共刺激后IL-8和IP-10的mRNA及蛋白表达对照组及LPS组升高,差异有统计学意义。不同浓度poly(I∶C)(0.001、0.01、0.1μg/mL)刺激后IL-8、IP-10 mR-NA和蛋白表达呈浓度依赖性升高;各组分别加入10μg/mLLPS共刺激后IL-8、IP-10 mRNA及蛋白表达未见进一步升高。③地塞米松(1μmol/L)及SB203580(20μmol/L)分别预处理30 min后给予0.1μg/mL poly(I∶C)及10μg/mL LPS共刺激,两组IL-8、IP-10 mRNA及IL-8、IP-10蛋白表达较共刺激组和共刺激+DMSO组降低,差异有统计学意义(P<0.01和P<0.05),其中地塞米松的抑制效应强于p38MAPK抑制剂。结论:poly(I∶C)和LPS共刺激能够诱导气道上皮细胞趋化因子的表达,poly(I∶C)能加重LPS所致气道上皮细胞趋化因子IL-8的表达。糖皮质激素及p38MAPK抑制剂具有抑制模拟病毒感染及内毒素诱导气道上皮细胞趋化因子表达,提示两者在病毒诱导的AECOPD中具有潜在治疗价值。AIM： To investigate the effect of co-stimulation of mimic viral infection [ potyinosinic, polycytidylic acid, poly （I：C） ] and endotoxin （lipopolysaccharide, LPS） on chem- otactic factors production of human bronchial epithelial cells and explore its related mechanism. METHODS： Human bronchial epithelial cells （16HBE） were challenged by co- stimulation of different concentrations of poly （ I ： C ） and LPS. Some other 16HBE cells, before the co-stimulation of poly（l： C） and LPS, were pretreated with dexamethasone and p38MAPK specific inhibitor （SB203580）, respectively. The levels of IL-g/CXCL8 and IP-10/CXCL] mRNA transcription were detected by RT-PCR after 6 h challenge. The contents of IL-8 and IP-]0 proteins were detected by ELISA after 24 h challenge. RESULTS： ① The mRNA and protein expression of IL-8, not IP-10, increased under 10 μg/mL LPS stimulation compared with control group. The mRNA and protein expressions of both IL-8 and IP-10 were elevated significantly by the co-stimulation of LPS and 0.1 μg/mL poly （I：C） compared with control group and sim- ple LPS groups. ②The mRNA and protein expressions of IL-8 and IP-10 increased under the challenge of different concentrations of poly（I：C） （0.001, 0.01, 0. 1μg/mL） in a concentration-dependent manner, which showed no change after adding 10 iag/mL LPS.③ Dexamethasone （1 μmol/L） and SB203580 （20 μmol/L） significantly decreased both the mRNA and protein production of IL-8 and IP-10 induced by co-stimulation of 0.1 μg/mL poly （I：C） and 10μg/mL LPS compared with the control group and control ＋ DMSO grouprespectively （ P 〈 0.01 and P 〈 0.05）. Inhibitive effect of dexamethasone was stronger than that of p38MAPK inhibi- tor. CONCLUSIONS： The co-stimulation of poly （ I： C） and LPS can induce chemokine expression of airway epithelial cells. Poly（I：C） enhances the IL-8 expression induced by single LPS challenge, which suggests that viral infection could enhance the infla

关 键 词：气道上皮细胞 聚肌胞苷酸 内毒素 IL-8 干扰素诱导蛋白10 

分 类 号：R392.11[医药卫生—免疫学] Q256[医药卫生—基础医学]