缺氧条件下大鼠神经元白细胞介素-1受体相关激酶-1的表达变化  

Hypoxia induces expression of interleukin-1 receptor-associated kinases-1 in rat neurons

作　　者：李江涛[1] 胡胜利[1] 杜鹏[1] 郑江[2] 缪洪平[1] 吴国材[1] 唐卫华[1] 刘智[1] 朱刚[1] 

机构地区：[1]第三军医大学西南医院,神经外科,全军神经系统微创诊治专科中心,重庆400038 [2]第三军医大学西南医院,神经外科,综合实验研究中心,重庆400038

出　　处：《第三军医大学学报》2010年第8期794-797,共4页Journal of Third Military Medical University

基　　金：重庆市自然科学基金(CSTC2007BA5010)~~

摘　　要：目的观察大鼠神经元B35细胞株中白细胞介素-1受体相关激酶-1(interleukin-1 receptor-associated kina-ses-1,IRAK-1)mRNA及蛋白在缺氧后表达量的变化,并探讨其变化与神经元中TNF-α表达的关系。方法将培养的B35细胞置于低氧(3%O2、5%CO2、92%N2)条件下培养1、3、6、12、24、48、72 h,应用RT-PCR方法检测IRAK-1 mRNA表达的变化;Western blot检测IRAK-1蛋白含量的变化;激光扫描共聚焦观察IRAK-1在细胞内的分布及含量变化;酶联免疫吸附法检测培养液上清肿瘤坏死因子-α(TNF-α)的含量,并与IRAK-1蛋白进行相关分析。结果B35细胞IRAK-1 mRNA表达与正常对照相比,1 h开始增加[(0.786±0.042),P<0.05],3 h增加明显[(0.947±0.055),P<0.05],6 h达到高峰(1.245±0.077),P<0.05],12 h仍维持在较高水平[(0.911±0.041),P<0.05],24 h降至正常水平以下但无明显差异[(0.596±0.047),P>0.05],48 h以后进一步减少[(0.520±0.049),P<0.05]。IRAK-1蛋白表达变化也呈类似趋势。相关性分析结果显示,培养上清液中TNF-α含量的变化与IRAK-1蛋白表达变化呈正相关(r=0.88,P<0.05)。结论神经元内存在TLR/IL-1R家族信号转导系统的关键因子IRAK-1,缺氧损伤可诱导其表达,与TNF-α的表达相关。Objective To investigate the dynamical changes of interleukin-1 receptor-associated kinases-1（IRAK-1） mRNA and protein,and to explore the relationship between these changes and concentrations of TNF-α in rat B35 neurons under hypoxia.Methods B35 cells exposed to hypoxia of 3%O2,5%CO2,and 92%N2 were cultured for 1,3,6,12,24,48 and 72 h respectively,the mRNA and protein expressions of IRAK-1 were detected by RT-PCR and Western blot analysis,the distribution of IRAK-1 in the cells were observed by laser scanning confocal microscopy（LSCM）,and the concentrations of TNF-α were measured by ELISA.Results The mRNA and protein expressions of IRAK-1 were increased after hypoxia for 1 h and decreased gradually after reaching the highest level at 6 h（P〈0.05）,so did the fluorescence intensity under LSCM and the concentrations of TNF-α.After 48 h,the concentrations of mRNA and protein were decreased lower than that in normoxia（P〈0.05）.The concentrations of TNF-α were correlated positively with the expression of IRAK-1 protein（r=0.88,P〈0.05）.Conclusion Hypoxia up-regulates the mRNA and protein of IRAK-1,which are correlated positively with the expression of TNF-α in rat B35 neurons.

关 键 词：白细胞介素-1受体相关激酶-1 缺氧 神经元 

分 类 号：R331.8[医药卫生—人体生理学] R345[医药卫生—基础医学]