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目的 :研究特异性p3 8分裂原激活的蛋白激酶 (MAPK)抑制剂SB2 0 3 580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 (KCl 5mmol/L)培养基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p3 8MAPK抑制剂SB2 0 3 580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。此保护作用具有浓度依赖性。培养于低钾环境中的神经元 ,其c Jun的表达和磷酸化水平升高 ,且激活了JNK的活性。当小脑颗粒神经元生长在含SB2 0 3 580 2 5μmol/L的低钾培养基中 ,c Jun的表达、磷酸化水平和JNK的活性都明显的降低。结论 :SB2 0 3 580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。
AIM: To investigate the effect and mechanism of a specific p38 MAPK inhibitor SB2 0 3 580 on hypocalcemia-induced cerebellar granule neuronal apoptosis. METHODS: JNK (c Jun N-terminal kinase) activity was measured in vitro by neuronal cultures, gel electrophoresis and the SAPK / JNK assay cassette. Results: KCl 5mmol / L induced apoptosis of cerebellar granule neurons with typical morphological and biochemical characteristics. However, the specific p3 8 MAPK inhibitor SB2 0 3 580 promoted the survival of cultured cerebellar granule neurons in hypokalemia by inhibiting apoptosis. This protective effect is concentration-dependent. Neurons cultured in hypokalemia have elevated c Jun expression and phosphorylation, and activate JNK activity. When cerebellar granule neurons grew in low potassium medium containing SB203 580 25μmol / L, c Jun expression, phosphorylation and JNK activity were significantly reduced. Conclusion: SB2 0 3 580 may protect cerebellar granule neurons from hypokalemia by inhibiting the activity of JNK and decreasing the phosphorylation of c Jun.