核因子-κB和p38丝裂原活化蛋白激酶调节脂多糖诱导的气道上皮细胞白细胞介素8的表达  

Induction of interleukin-8 by lipopolysaccharide in human airway epithelial cells via NF-κB and p38 mitogen-activated protein kinase

作　　者：李亚清[1] 严建平[1] 许武林[1] 王宏[1] 

机构地区：[1]浙江省人民医院呼吸内科,浙江杭州310014

出　　处：《中国药理学通报》2011年第2期182-186,共5页Chinese Pharmacological Bulletin

基　　金：浙江省医药卫生科学研究基金资助项目(No2008B007);浙江省人民医院博士科研基金经费资助(No200701)

摘　　要：目的探讨脂多糖(LPS)对HBE4-E6/E7细胞IL-8表达的影响及Toll样受体4(toll-like receptor 4,TLR-4)、p38MAPK、NF-κB在其中的作用。方法在量效实验中,分别以0、0.1、1、10、100μg.L-1 LPS刺激HBE4-E6/E7细胞24 h;在时效实验中,以10μg.L-1 LPS分别刺激HBE4-E6/E7细胞0、8、12、16、24 h。以100μg.L-1 TAK-242、10 mg.L-1SB202190、50 mg.L-1 PDTC预处理HBE4-E6/E7细胞,以100μg.L-1 LPS刺激24 h。半定量RT-PCR法检测IL-8mRNA表达;酶联免疫吸附试验检测IL-8蛋白水平;凝胶阻滞分析实验检测NF-κB活性。结果 HBE4-E6/E7细胞IL-8 mRNA的表达随LPS刺激剂量的增加明显增加(P<0.05);在24 h内,以10μg.L-1 LPS刺激时,IL-8 mRNA的表达随刺激时间的延长明显增加(P<0.05)。TAK-242、SB202190和PDTC均能明显抑制LPS诱导的HBE4-E6/E7细胞IL-8 mRNA和蛋白的表达(P<0.05)。TAK-242和PDTC均能明显抑制LPS诱导的NF-κB的活性(P<0.05);SB202190不能抑制LPS诱导的NF-κB活性(P>0.05)。结论 LPS能诱导HBE4-E6/E7细胞IL-8的表达,TLR4、p38MAPK、NF-κB均参与调控LPS诱导的气道上皮细胞IL-8的表达,且p38 MAPK是通过不依赖NF-κB信号途径来参与调节的。Aim To study the effect of lipopolysaccharide（LPS）on interleukin（IL）-8 expression in HBE4-E6/E7 cells,and explore the role of Toll-like receptor-4（TLR-4）,NF-κB and p38 mitogen-activated protein kinase（MAPK）in IL-8 expression induced by LPS.Methods HBE4-E6/E7 cells were stimulated for 24 h by different concentrations of LPS（0,0.1,1,10,100 μg·L-1） and were stimulated for 0,8,12,16,24 h by 10 μg·L-1 LPS.HBE4-E6/E7 cells were incubated for 30 min with 100 μg·L-1 TAK-242,10 mg·L-1 SB202190,50 mg·L-1 PDTC,and then stimulated for 24 h by 100 μg·L-1 LPS.Expression levels of IL-8 were detected respectively by semi-quantitative reverse transcription-polymerase chain reaction（RT-PCR）and enzyme-linked immunosorbent assay（ELISA）.NF-κB activity was detected via electrophoretic mobility shift assay（EMSA）.Results IL-8 expression induced by LPS in HBE4-E6/E7 cells were markedly increased in both a dose-dependent manner and a time-dependent manner（P0.05）.TAK-242,PDTC and SB202190 can inhibit IL-8 expression induced by LPS in HBE4-E6/E7 cells（P0.05）.TAK-242 and PDTC can suppress NF-κB activity induced by LPS（P0.05）,while SB202190 can not（P0.05）.Conclusions Induction of IL-8 by LPS in HBE4-E6/E7 cells may be regulated by NF-κB and p38 MAPK signal pathway.

关 键 词：Toll样受体4 白细胞介素8 核因子-KAPPAB P38 气道上皮细胞 

分 类 号：R322.33[医药卫生—人体解剖和组织胚胎学] R329.24[医药卫生—基础医学]