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目的 用星形孢菌素诱导 NG1 0 8-1 5细胞损伤建立凋亡模型 ,观察乙酰胆碱酯酶 (ACh E)抑制剂他克林是否具有抗凋亡作用。方法 LDH活性测定判断细胞损伤状况 ,DNA染料 Hoechst3 3 3 42染色观察细胞核形态 ,SDS-PAGE和 Western blot印迹分析 Bax及 Bcl-2的蛋白表达水平。结果 NG1 0 8-1 5细胞经 0 .1μmol/ L星形孢菌素处理 2 4h,细胞内大量 LDH释放至培养液中 ,用 0 .1 mmol/ L 他克林预处理 2 h可使 LDH释放量由 45 %减至 3 2 % (P<0 .0 1 )。他克林预处理能延迟和减少星形孢菌素诱导的 Bax表达的上调 ,同时增加了星形孢菌素诱导 1 2~ 2 4h后 Bcl-2的表达水平。结论 他克林预处理对星形孢菌素诱导的凋亡性损伤有显著保护作用 ,可能通过抑制或延迟 Bax表达及促进 Bcl-2表达来发挥神经保护作用
OBJECTIVE To study the apoptosis of NG180-151 cells induced by staurosporine and to observe whether tacrine, an inhibitor of acetylcholinesterase (AChE), has an anti-apoptotic effect. Methods LDH activity was measured to determine the cell damage status. The nuclear morphology was observed by DNA stain Hoechst3 3 42 staining, and the protein expression of Bax and Bcl-2 was analyzed by SDS-PAGE and Western blot. Results NG1 0 8-1 5 cells were treated with 0 .1 μmol / L staurosporine for 24 h, and a large amount of LDH was released into the culture medium. Pretreatment with 0 .1 mmol / L tacrine for 2 h made LDH Release from 45% to 32% (P <0.01). Tacrine pretreatment delayed and decreased staurosporine-induced upregulation of Bax expression and increased the level of Bcl-2 expression by staurosporine 12-24 h after induction. Conclusion Tacrine pretreatment significantly protects against staurosporine-induced apoptotic injury and may play a neuroprotective role by inhibiting or delaying Bax expression and promoting Bcl-2 expression