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目的 :研究脑缺血损伤诱导非受体酪氨酸蛋白激酶Src之丝、苏、酪氨酸残基磷酸化及其与Pyk2结合的变化并探讨其可能的调节机制。方法 :四动脉结扎模型诱导大鼠前脑缺血损伤 ,免疫沉淀和免疫印迹法观察Src氨基酸残基磷酸化及其与Pyk2结合的变化。 结果 :缺血 15min后复灌 ,Src的Tyr 4 16和Ser而不是Thr的磷酸化有明显增加 ,且复灌 6hTyr 4 16磷酸化升至最高 ,而Ser磷酸化 1h升至顶峰 ,它们分别是正常组的 2 .8和 2 .3倍 ;此外 ,缺血 /复灌明显诱导Src与Pyk2的结合 ,相似于Tyr 4 16磷酸化变化 ,复灌 6h升至最高。 5 0mg/kg的氯胺酮能明显抑制缺血 /复灌诱导的SrcTyr 4 16磷酸化及其与Pyk2结合增加 ,与对照组比有显著性差异 (P <0 .0 5 )。结论 :脑缺血诱导了Src蛋白激酶Ser和Tyr 4 16磷酸化及其与Pyk2结合 ,它们参与了Src的激活和NMDA受体功能的自调。
OBJECTIVE: To investigate the changes of phosphorylation of silk fibroin, tyrosine and tyrosine residues in non-receptor tyrosine kinase kinases induced by cerebral ischemia and their association with Pyk2, and to explore the possible regulatory mechanism. Methods: The model of four-artery ligation was used to induce the forebrain ischemic injury in rats. The phosphorylation of Src amino acid residues and the binding to Pyk2 were observed by immunoprecipitation and immunoblotting. Results: Phosphorylation of Tyr 4 16 and Ser but not Thr in Src increased significantly after reperfusion 15 min after ischemia, and phosphorylation of 6 th Tyr 4 16 increased to the highest after reperfusion, whereas Ser phosphorylation peaked at 1 h In addition, ischemia / reperfusion significantly induced the binding of Src to Pyk2, which was similar to the change of phosphorylation of Tyr 4 16, and reached the highest after reperfusion for 6 hours. Ketamine at a dose of 50 mg / kg significantly inhibited phosphorylation of SrcTyr 4 16 and increased binding to Pyk2 after ischemia / reperfusion, which was significantly lower than that of the control group (P <0.05). CONCLUSION: Cerebral ischemia induces the phosphorylation of Src protein kinase Ser and Tyr 4 16 and their binding to Pyk2, which are involved in the activation of Src and the self-regulation of NMDA receptor function.