转录因子NF-kB在肿瘤坏死因子α介导的胰岛β细胞凋亡中的作用  

Role of nuclear factor of kB in tumor necrosis factor α-mediated INS-1 cell apoptosis

作　　者：杨立勇[1] 杨永年[2] 陈志坚 

机构地区：[1]福建医科大学附属第一医院内分泌科,350005 [2]复旦大学附属中山医院内分泌科 [3]USA University of Texas southwestern Medical Center,752359148

出　　处：《中华医学杂志》2003年第8期688-690,共3页National Medical Journal of China

基　　金：福建省自然科学基金资助项目 (C0 110 0 12 )

摘　　要：目的　探讨肿瘤坏死因子 (TNFα)是否可引起胰岛 β细胞凋亡以及转录因子核因子kB(NF kB)在TNFα诱导的 β细胞凋亡中所起的作用。 方法　培养INS 1胰岛 β细胞 ,采用DNA重组、转染和再感染技术获得可表达NF kB的特异性抑制物IkBα的突变体IkBαΔN的INS 1/IkBΔN细胞。应用DNA片段分析技术和kB luc报告基因荧光分析技术观察NF kB活性和 β细胞的凋亡情况。 结果　IL Iβ可诱导INS 1细胞的NF kB激活 ,但在INS 1/IkBΔN细胞则无此作用。将细胞与IL 1β( 10 μg/L)、TNFα( 10 0 μg/L)以及IFNγ( 10 0 0 0 0U/L)一道培养 48h后显示 ,TNFα与IFNγ合用可诱导INS 1细胞发生凋亡 ,而在INS 1/IkBΔN细胞未见此现象。单用IL Iβ或IFNγ或IL Iβ加IFNγ均未能诱导INS 1细胞凋亡。结论　凋亡是TNFα导致β细胞死亡的方式之一 ,NF kB在TNFα介导的β细胞凋亡中具有重要作用 ,抑制NF kB激活可能保护 β细胞免于TNFα诱导的凋亡。Objective To investigate the effect of cytokine tumor necrosis factor α(TNFα) on pancreatic β cells apoptosis and the effect of transcription factor nuclear factor of kappa B (NF-kB) on TNFα-mediated apoptosis. Methods INS-1 cells, a pancreatic β cell line, were cultured,and recombinant DNA technique, transfection and reinfection technology were used to obtain INS-1/ IkBΔN cells expressing inhibitor of kB (IkB)αΔN, mutant IkBα (inhibitor of NF-kB). DNA fragmentation assay and fluorescence analysis using a kB-luc reporter gene were applied to observe the NF-kB activity and β cell apoptosis. Results NF-kB activity induced by IL-Iβ was detectable in INS-1 cells but not in INS-1/ IkBΔN cells. After incubation of the cells with IL-1β (10 μg/L),TNFα(100 μg/L) and interferon (IFN)γ (100 000 U/L) for 48 hours, the combination of TNFα and IFNγ induced apoptosis in INS-1 cells but not in INS-1/IkBΔN cells. No apoptosis was observed after incubation of INS-1 cells with IL-1β or IFNγ or IL-1β plus IFNγ . Conclusion Apoptosis is one of the TNFα-induced β-cell death forms. NF-kB may play an important role in the TNFα-mediated β-cell apoptosis. Inhibition of NF-kB activation protects β-cells from TNFα-induced apoptosis.

关 键 词：转录因子 NF—kB 肿瘤坏死因子Α 介导 胰岛Β细胞凋亡 脱噬作用 

分 类 号：R587[医药卫生—内分泌]