IκBα突变型基因提高过氧化氢处理后永生化星形胶质细胞的存活率  

IκBα Mutant Gene Improved the Survival of Immortalized Astrocytes after Hydrogen Peroxide Treatment

作　　者：祝畅[1] 桂伶俐[1] 姚文龙[1] 张传汉[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院麻醉科,武汉430030

出　　处：《神经损伤与功能重建》2013年第5期322-325,共4页Neural Injury and Functional Reconstruction

基　　金：国家自然科学基金资助项目(No.81100919)

摘　　要：目的:探讨IκBα突变型基因对永生化星形胶质细胞株(IAST)NF-κB的活性及过氧化氢损伤后细胞存活率的影响。方法:通过脂质体法将携带IκBα突变型基因及对照载体转入IAST,通过Western Blot检测IκB蛋白的表达,用荧光素酶报告基因检测细胞中NF-κB的活性,用MTT法分别检测0.1 mmol/L、0.2 mmol/L、0.4 mmol/L过氧化氢处理2 h、4 h后转IκBα突变型基因及对照载体IAST的细胞存活率,免疫细胞化学法检测0.4 mmol/L过氧化氢处理4 h前后两组细胞中p65的表达。结果:转IκBα突变型基因的IAST中可见突变型IκBα表达,荧光素酶催化底物所发出的荧光强度较低(P<0.05)。各浓度过氧化氢处理4 h后转IκBα突变型基因的IAST细胞存活率较转对照载体的IAST高(P<0.05)。0.4 mmol/L过氧化氢处理4 h后转IκBα突变型基因及对照载体IAST中p65的表达出现核内移。结论:IκBα突变型基因可降低IAST中NF-κB的活性,提高过氧化氢处理后该细胞株的存活率。Objective： To study the effect of mutated IKBα gene on the NF-KB activity and cell viability of immortalized astrocyte strain （lAST） after hydrogen peroxide injury. Methods： Eukaryotic expression vector car- fled I KBα mutant gene （pcDNA3.1/IKBαM） or control empty vector （pcDNA3.1） was transfected into lAST by li- posome method. IKB protein expression was detected by Western Blot and NF-KB activity was visualized by lu- ciferase reporter gene with NF-KB-specific promoter. The cell viability of IKBα mutant gene or control vector transfected lAST was detected by MTT method after treatment by 0.1 mmol/L, 0.2 mmol/L or 0.4 mmol/L hydro- gen peroxide for 2 h or 4 h. The expression of p65 in IKBα mutant gene or control vector transfected lAST was detected by immunocytochemistry before or after the treatment by 0.4 mmol/L hydrogen peroxide for 4 h. Results： The specific band of the mutant IKBα was visible in pcDNA3. I/IKBccM transfected lAST by Western blot. The intensity of the fluorescence emitted by luciferase reporter gene in pcDNA3.1/IKBαM transfected lAST was lower than pcDNA3.1 transfected lAST （P〈0. 05）. The cell survival rate ofpcDNA3.1/IKBαM transfected lAST was higher than pcDNA3.1 transfected lAST after Various concentrations of hydrogen peroxide treatment for 4 h （P〈0.05）. The p65 expression in the pcDNA3.1/IKBαM or pcDNA3.1 transfected lAST was translocated to the nuclear after 0.4 mmol/L hydrogen peroxide treatment for 4 h. Conclusion： IKBα mutant gene can reduce the activity of NF-KB in lAST, and improve the survival rate of the cell line after the hydrogen peroxide treat- ment.

关 键 词：星形胶质细胞 基因表达 NF-κB抑制因子α 过氧化物 

分 类 号：R741[医药卫生—神经病学与精神病学]