Cursio Raffaele, Miele Claudia, Filippa Nathalie, Colosetti Pascal, Auberger Patrick, Van Obberghen Emmanuel, Gugenheim Jean
Laboratoire de Recherches Chirurgicales, IFR 50, Faculté de Médecine, Université de Nice Sophia Antipolis, Nice, France.
Langenbecks Arch Surg. 2009 Jan;394(1):123-31. doi: 10.1007/s00423-008-0394-3. Epub 2008 Aug 5.
Phosphoregulation of signal transduction pathways is a complex series of reactions that may modulate the cellular response to ischemia-reperfusion (I-R). The aim of this study was to evaluate the effect of normothermic liver I/R-induced apoptosis on phosphorylation and activation of signal proteins in tyrosine kinase pathways.
In rats, a segmental normothermic ischemia of the liver was induced for 120 min. Liver apoptosis was determined using terminal deoxynucleotide-transferase-mediated deoxyuridine triphosphate nick end labeling assay, and activity of caspases-3 and -7 was determined by fluorescence. Liver tyrosine phosphorylation of proteins was examined by Western blot analysis.
Normothermic I-R resulted in increased in vivo caspases-3 and -7 activity and in liver apoptosis. Shc tyrosine phosphorylation and activation of ERK1/2 were increased after reperfusion, while tyrosine phosphorylation of IRS-1 and activation of PKB/Akt were decreased.
Normothermic liver I-R leads to increased apoptosis and to modifications in protein tyrosine phosphorylation pathways.
信号转导通路的磷酸化调节是一系列复杂的反应，可能会调节细胞对缺血再灌注（I-R）的反应。本研究的目的是评估常温肝脏I/R诱导的细胞凋亡对酪氨酸激酶通路中信号蛋白磷酸化和激活的影响。
在大鼠中，诱导肝脏节段性常温缺血120分钟。使用末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸缺口末端标记法测定肝脏细胞凋亡，并通过荧光法测定半胱天冬酶-3和-7的活性。通过蛋白质印迹分析检测肝脏蛋白质的酪氨酸磷酸化。
常温I-R导致体内半胱天冬酶-3和-7活性增加以及肝脏细胞凋亡。再灌注后，Shc酪氨酸磷酸化和ERK1/2激活增加，而IRS-1的酪氨酸磷酸化和PKB/Akt激活减少。
常温肝脏I-R导致细胞凋亡增加以及蛋白质酪氨酸磷酸化途径的改变。
