【摘要】 目的 :研究化学合成的蛋白激酶抑制剂Olomoucine对低钾诱导的小脑颗粒神经元凋亡的作用及机制。方法 :体外神经元培养、凝胶电泳和SAPK/JNK分析盒测定JNK(c -Jun氨基末端激酶 )活性。结果 :低钾 (KC15mmol/L)诱导小脑颗粒神经元的具有典型形态学和生化特征的凋亡。但是 ,特异性的细胞周期蛋白依赖性激酶 (CDKs)抑制剂Olomoucine通过抑制凋亡 ,而促进低钾环境中培养的小脑颗粒神经元的存活 ,且此保护作用具有浓度依赖性。因激活了JNK的活性而升高了c -Jun的磷酸化水平 ,导致培养于低钾环境中的神经元凋亡。但是 ,当小脑颗粒神经元生长在含Olomoucine 5 0 μmmol/L的低钾培养基中 ,c -Jun的磷酸化水平和JNK活性都明显降低。结论 :Olomoucine可能通过抑制JNK的活性 ,降低c -Jun的磷酸化水平而对低钾培养的小脑颗粒神经元具有保护作用更多还原

【Abstract】 To study the effect of the specific cyclin dependent kinases (CDKs) inhibitor Olomoucine on apoptosis induced by low potassium and its mechanisms by using agarose gel eletrophoresis and SAPK/JNK assay kit rto measure SAPK/JNK activity. The results showed that switching cerebellar granule neurons from depolarizing conditions (KCl 25 mmol/L) to non-depolarizing conditions (KCl 5 mmol/L,cLK)exhibited the morphological and biochemical features characteristic of apoptosis. But the specific CDKs inhibitor Olomoucine promoted the survival of cerebellar granule neurons cultured in cLK medium by blocking apoptosis. This protective action was in a concentration-dependent manner. The phosphorylation of c-Jun increased and the activity of c-Jun N-terminal protein kinase(JNK) elevated when cerebellar granule neurons were cultured in cLK medium. But when the cerebbellqr granule neurons cultured in cLK medium were exposed to Olomoucine 50μmmol/L, the phosphorylation of c-Jun and the activity of JNK all decreased evidently. It indicated that Olomoucine inhibited the activation of JNK and decreased the phosphorylation of c-Jun to protect granule neuons from apoptosis induced by low potassium.更多还原