缺氧大鼠神经元中IRAK-4的表达  

Experimental study of IRAK-4 expression in hypoxic neurons of rats

作　　者：李江涛[1] 胡胜利[1] 胡军[1] 杜鹏[1] 冯华[1] 杨云峰[1] 魏林节[1] 朱刚[1] 

机构地区：[1]第三军医大学附属西南医院神经外科,重庆400038

出　　处：《中华创伤杂志》2010年第7期648-652,共5页Chinese Journal of Trauma

基　　金：重庆市自然科学基金重点资助项目（2007BA5010）

摘　　要：目的 探讨大鼠神经元缺氧后IL-1受体相关激酶-4（interleukin-1receptor-associated kinases-4,IRAK-4）的表达及其在调节炎症反应中的作用.方法 将培养的B35细胞置于低氧（3%O2,5%CO2,92%N2）条件下分别培养1,3,6,12,24,48,72和96 h,用RT-PCR及Western blot检测IBAK-4的mRNA及蛋白表达的变化,激光扫描共聚焦显微镜观察IRAK-4在细胞内的表达变化情况,酶联免疫吸附法检测培养液上清IL-6的含量.结果 B35细胞IRAK-4的mRNA和蛋白表达随缺氧时间延长而变化,1 h开始增加,6 h达到高峰（P〈0.05）,12 h仍维持在较高水平（P〈0.05）,自24 h下降至常氧水平（P〉0.05）,48 h以后降低至常氧水平以下（P〈0.05）.IRAK-4的免疫荧光结果显示,随着缺氧时间延长荧光强度逐渐增强.培养上清液中IL-6含量的变化与IRAK-4蛋白表达变化呈正相关（r=0.84,P〈0.05）.结论 缺氧在一定时间内可以促使大鼠神经元IRAK-4在转录水平和翻译水平的表达增加,并可能参与调控下游炎症反应,使IL-6的表达增加.Objective To investigate the dynamic changes of interleukin-1 receptor-associated kinases （IRAK-4） in hypoxic neurons and explore their role in regulation of inflammatory reaction. Methods The B35 cells exposed to hypoxia of 3% O2,5% CO2 and 92% N2 were cultured for 1,3,6, 12,24,48,72 and 96 hours respectively. Then, mRNA and protein expressions of IRAK-4 were detected by RT-PCR and Western blot, the expression of IRAK-4 in the cells were observed by laser scanning con-focal microscope （LSCM）, and the concentration of IL-6 was measured by ELISA method. Results After hypoxia, the mRNA and protein expressions of IRAK-4 were increased at one hour, reached the peak at six hours （P〈0.05）, kept at a high level at 12 hours （P〈0.05） , but decreased gradually to the normal oxygen level at 24 hours （P 〈 0.05） and to below the normal oxygen level at 48 hours （P 〈 0.05）. Immunofluorescence results showed that the fluorescence intensity of IRAK-4 was gradually increased with time. The changes of IL-6 in the supernatant were positively correlated with protein expression of IRAK-4 （r =0.84, P 〈 0.05 ）. Conclusions Hypoxia can increase the expression of IRAK-4 at transcription and translation levels in a certain period of time, which may participate in down-stream inflammatory reaction and lead to increase of IL-6 expression.

关 键 词：缺氧 白细胞介素-1 神经元 继发性脑损伤 

分 类 号：R686[医药卫生—骨科学]