Essential role for nuclear factor κB in ischemic preconditioning for ischemia-reperfusion injury of the mouse liver
Essential role for nuclear factor κB in ischemic preconditioning for ischemia-reperfusion injury of the mouse liver
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DOI：
10.1097/00007890-200208270-00021
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发表时间：
2002-08-27
期刊：
影响因子：
6.2
通讯作者：
Miwa, K
中科院分区：
文献类型：
--
作者：
Funaki, H;Shimizu, K;Miwa, K
背景缺血预处理可保护多种器官免受缺血损伤，但其确切机制尚不清楚。为了探讨缺血预处理发挥其保护作用的分子机制，我们检测了转录活性。因子核因子（NF）-κ B和随后的炎症基因表达。在脾皮下移位后，将小鼠用于全肝缺血再灌注实验。小鼠肝脏缺血70分钟，然后再灌注规定的时间。在缺血70 min前进行缺血预处理，即缺血V min再灌注20 min。通过电泳迁移率变动分析、蛋白质和酪氨酸磷酸化分析NF-κ B活性。用Western blot分析抑制因子κ B-α的水平，用半定量逆转录聚合酶链反应分析肿瘤坏死因子（TNF）-α和细胞间粘附分子1 m-RNA水平。NF-κ B在再灌注后30 min内即被激活，并持续激活4 h。缺血预处理可减弱随后长时间缺血再灌注后NF-κ B的激活，同时降低TNF-α和细胞间粘附分子1 mRNA的表达，前者有统计学意义（P
Background. Ischemic preconditioning protects various organs from subsequent ischemic insult, but the precise mechanisms underlying this phenomenon re main undefined. To investigate the molecular mechanism by which ischemic preconditioning exerts it protective effect, we examined the activity of the transcription. factor nuclear factor (NF)-kappaB and subsequent inflammatory gene expression.Methods. Mice were used for total hepatic ischemia reperfusion experiments after subcutaneous transposition of the spleen. Mouse liver was subjected to ischemia for 70 min followed by reperfusion for defined times. Ischemic preconditioning that consisted of V min of ischemia and 20 min of reperfusion was per formed before 70 min of ischemia. NF-kappaB activity was analyzed by electrophoretic mobility shift assay, an the protein and tyrosine phosphorylation. levels of in hibitor kappaB-alpha were assessed by Western blot analysis Semiquantitative reverse-transcriptase polymerase chain reaction was used to analyze tumor necrosis factor (TNF)-alpha and intercellular adhesion molecule 1 m-RNA levels.Results. NF-kappaB was activated within 30 min after initiation of reperfusion and remained activated for 4 hr. Ischemic preconditioning attenuated NF-kappaB activation after subsequent prolonged ischemia and reperfusion and simultaneously decreased the expression of TNF-alpha and intercellular adhesion molecule 1 mRNA, the former statistically significantly (P