PKC/MAPK/NF-κB信号通路在低氧诱导大鼠单核细胞表达IL-1β中的作用  

Role of PKC/MAPK/NF-kappa B signal cascade in expression of IL-1β in rat monocytes exposed to hypoxia

作　　者：徐智[1] 吴国明[1] 李昆霖[2] 钱桂生[1] 王兴胜[2] 陈维中[1] 

机构地区：[1]解放军第三军医大学新桥医院全军呼吸内科研究所,400037 [2]解放军第三军医大学大坪医院全军呼吸科,400037

出　　处：《中国急救医学》2008年第9期803-807,共5页Chinese Journal of Critical Care Medicine

基　　金：国家自然科学基金(No.30500230)

摘　　要：目的研究PKC／MAPK／NF—κB信号通路在低氧诱导大鼠外周血单核细胞（peripher—al blood monouclear cells，PBMCs）白细胞介素-1β（IL-1β）表达中的作用，探讨低氧与全身炎症反应综合征（SIRS）的关系，为进一步研究老年多器官功能障碍综合征（MODSE）的肺启动机制奠定基础。方法采用明胶法分离大鼠外周血单核细胞，分为低氧组和Chelerythrine＋低氧组。Chelerythrine＋低氧组于低氧前予10 μmol／L Chelerythrine预处理。然后两组细胞均于低氧条件下（3％O2，5％CO2，92％N2）培养0、1、3、6、9、12、24h后，收集细胞及培养液上清，分别采用PKC、MAPK活性检测试剂盒、电泳迁移分析法（EMSA）、逆转录PCR（RT—PCR）检测PKC、MAPK、NF—κB活性及IL-1β表达量。结果低氧1～9h，PKC、MAPK活性，NF—κB结合活性及IL-1β表达量显著高于正常对照组（P〈0．01）。低氧后1～24h，PKC、MAPK活性、NF—κB结合活性与IL-1β mRNA表达水平间呈显著正相关（P〈0．01—0．05）。10 μLmol／L Chelerythrine可显著抑制低氧诱导的PKC、MAPK、NF—κB活性升高和IL-1β表达。结论低氧可显著增强大鼠外周血单核细胞的PKC、MAPK活性及NF—κB结合活性，并可诱导其产生大量的促炎症因子IL-1β，这些变化可能与急性呼吸窘迫综合征（acuterespiratorydysfunction，ARDS）时血浆中大量促炎症因子持续存在密切相关。Objective To investigate the role of PKC/MAPK/NF - κB signal cascade in the expression of IL - 1β in rat peripheral blood monouclear cells （ PBMCs） exposed to hypoxia, and explore the relationship between hypoxia and system inflammation response syndrome （ SIRS） for the further study of the primordial role of lung in the pathogenesis of the multiple organ dysfunction syndrome in the elderly （MODSE）. Methods Purified rat PBMCs were randomly divided into three groups： hypoxia group and Chelerythrine ＋ hypoxia group. The Chelerythrinc ＋ hypoxia group were pretreated with 10 μLmol/L Chelerythrine before being exposed to hypoxia. Then two groups were exposed to hypoxia （ 3% 02 , 5% CO2, 92% N2） for 0,1,3,6,9,12,24 h. The PKC and MAPK activity were assayed by PKC kit and MAPK kit respectively. The NF - κB binding activity was detected by electrophoretie mobility shift assay （EMSA）. The expression of IL- 1β was detected by reverse transcriptase PCR （RT- PCR）. Results The activities of PKC, MAPK and NF - κB and the expression of IL - 1β increased significantly during 1 - 9 h of hypoxia （ P 〈 0. 01 ）. There were significant positive correlations between the activities of PKC, MAPK and NF - κB and the expression of IL - 1β （ P 〈 0. 01 - 0. 05 ）. 10 μmol/L Chelerythrine suppressed the hypoxia - induced PKC, MAPK and NF - κB activation and the expression of IL - 1β. Conclusion Hypoxia enhances the activities of PKC, MAPK and NF - κB in rat PBMCs, and up - regulates the expression of IL - 1β in mRNA and protein levels subsequently. These changes might be one of the mechanisms of a mass of pro - inflammatory factors exist persistently in blood plasma in acute respiratory distress syndrome （ARDS） patients.

关 键 词：低氧 蛋白激酶C 丝裂原活化蛋白激酶 核转录因子-ΚB 白细胞介素-1β 

分 类 号：R364.4[医药卫生—病理学] R392.12[医药卫生—基础医学]