神经调节蛋白1β在白细胞介素1β所致气道黏液高分泌中的作用  

Role of neuregulin-1β in airway hypersecretion induced by in interleukin-1β

作　　者：周佳[1] 尤列·皮尔曼[2] 维克多·科罗索夫[2] 周向东[1] 

机构地区：[1]重庆医科大学附属第二医院呼吸内科,重庆400010 [2]俄罗斯医学科学院远东呼吸生理与病理中心

出　　处：《中华医学杂志》2012年第28期1988-1991,共4页National Medical Journal of China

基　　金：国家自然科学基金，国家自然科学基金委员会批准资助对外交流与合作项目

摘　　要：目的探讨神经调节蛋白1p（NRG-1p）在白细胞介素1β（IL-1β）诱导的气道黏液高分泌中的作用。方法用IL-1β刺激人气道上皮细胞株HBEl6细胞构建黏液高分泌模型，逆转录PCR法检测NRG-1βmRNA和黏蛋白（MUC）5ACmRNA，ELISA法检测NRG-1β和MUC5AC蛋白水平。用NRG-1β刺激后，ELISA法检测MUC5AC表达，Western印迹法检测磷酸化人表皮生长因子受体（ErbB）1～4。预先分别给予ErbBl—4抗体及p38触分裂原活化蛋白（MAPK）特异性抑制剂、细胞外信号调节蛋白（ERK）1／2特异性抑制剂、丝裂原压力活性蛋白激酶（MSK）1特异性抑制剂、环磷酸腺苷（c—AMP）反应原件结合蛋白（CREB）单克隆抗体，再经NRG-1β刺激后，ELISA法检测MUC5AC蛋白表达。结果IL-113显著增加NRG-1β、MUC5ACmRNA水平及其蛋白表达，且在蛋白水平呈现浓度正相关性。单独给予NRG-1β（1、10、100、200nmol／L）刺激HBEl6细胞，MUC5AC蛋白表达（0．3284-0．055、0．364±0．086、0．650±0．134、0．586±0．068）较对照组（0．227±0．019）显著增加，差异均有统计学意义（均P〈0．05），同时磷酸化ErbB2、3呈阳性表达，而磷酸化ErbBl、4呈近似阴性表达。预先给予ErbB2、3抗体、p38MAPK特异性抑制剂、ERKl／2特异性抑制剂和MSKl抑制剂、CREB抗体再给予NRG-1β母刺激者MUC5AC蛋白表达（0．221±0．033、0．238±0．044、0．386±0．021、0．352±0．022、0．294±0．017、0．252±0．019）较单独给予NRG-1β（0．650±0．134）显著降低，差异均有统计学意义（均P〈0．05）。结论IL—β导致气道黏液高分泌可能通过NRG．1β／ErbB2、3异二聚体信号通路，并激活后续的MAPK／MsKl／cREB信号路径实现。Objective To explore the role of neuregulin 1β （NRG-1β） in airway hypersecretion induced by interleukin （IL）-1β. Methods After stimulating the airway epithelial cell line HBE16 with IL-1β, the expressions of NRG-1β mRNA and mucin （MUC） 5AC mRNA were detected by reverse transeription-polymerase chain reaction （ RT-PCR）, the proteins of NRG-1β and MUC5AC measured by enzyme-linked immunosorbent assay （ELISA） and phosphorylated erythroblastic leukemia viral oncogene homolog （ErbB）1 -4 detected by Western blot. The cells were pre-treated with antibodies of ErbB1-4, inhibitors of pβ8 mitogen-aetivated protein kinase （ MAPK）, ERK1/2, mitogen- and stress-activated protein kinase （MSK）1 and antibody of cAMP-response element-binding protein （CREB）. After the addition of stimulant NRG-1β, MUC5AC was measured by ELISA. Results IL-1β could increase markedly the levels of NRG-1β mRNA and MUC5AC mRNA and also the proteins of NRG-1β and MUC5AC in a dose-dependent fashion. NRG-1β at concentrations of 1, 10, 100, 200 nmoVL increased the expression of MUC5AC（0. 328 ~ 0. 055, 0. 364±0. 086, 0. 650±0. 134, 0. 586±0. 068） versus the control group （0. 227 ~ 0. 019）. And the results had statistical significances （P 〈 0. 05 ）. The expressions of phosphorylated ErbB2and ErbB3 stimulated by NRG-113 were positive while those of phosphorylated ErbB1 and ErbB 4 negative. After a pretreatment of antibodies of ErbB2, ErbB3, inhibitors of p38MAPK, ERK1/2, MSK1 and antibody of CREB and a stimulation of NRG-Iβ, the expression of MUC5AC decreased （0. 221±0. 033, 0. 238 ±0.044, 0.386 ±0.021, 0.352 ±0.022, 0.294 ~0.017, 0.252 ~0.019） versus the NRG-Iβ group （0. 650±0. 134）. And the results had statistical signifieanees （P 〈 0. 05 ）. Conclusion IL-1β may cause airway hyperseereton probably through the combination of NRG-1β with ErbB2 and ErbB3 heterodimers and the activation of MAPK/MSK1/CREB signal conduction.

关 键 词：神经调节蛋白1 白细胞介素1Β 黏蛋白类 受体 表皮生长因子 

分 类 号：R363[医药卫生—病理学]