烟碱拮抗秋水仙碱诱导的乳大鼠大脑皮质神经元凋亡

目的：探讨烟碱拮抗秋水仙碱诱导乳大鼠大脑皮质神经元凋亡的作用机制。方法: 培养Sprague-Dawley（SD）乳大鼠大脑皮层神经元，Hoechst33258核荧光染色进行凋亡率计数检测神经元凋亡, Akt（ser473）Western blotting分析试剂盒做激酶分析。结果: 0.1 μmol/L秋水仙碱可降低皮层神经元的存活率，诱导大鼠皮层神经元凋亡。但是，当10 μmol/Ｌ烟碱预孵2 h后再加入0.1 μmol/L秋水仙碱作用24 h, 烟碱可拮抗秋水仙碱的此种凋亡作用,将凋亡率从（62±1）％ 降低到 (38±2) ％,P<0.05。Akt（ser473）磷酸化在0.5 h时开始下降，于6 h时最低，仅为对照组的1/3。且Akt（ser473）磷酸化水平随时间（0.5、1、6、12和18 h）呈钟型性改变,分别是对照组的1.3、3.7、2.4、2.1和1.9倍,P<0.05。结论: 烟碱对大鼠皮质神经元凋亡的拮抗作用可能与其激活Akt有关。

Effect of nicotine against apoptosis of rat cortical neurons induced by colchicines

AIM:To explore the mechanism of nicotine against the apoptosis induced by colchicines in rat cortical neurons. METHODS:Cortical neurons were cultured from newborn Sprague-Dawley (SD) rats (less than 12 h). The rate of apoptosis was measured by Hoechst33258 fluorescence staining in the neurons, and the activity of Akt473 was analyzed by assay kit Akt473. RESULTS:The apoptosis of cortical neurons can be induced by 0.1 μmol/L colchicine. The phosphorlation of Akt 473 decreased significantly (1/3 times of the control group, P<0.01). However, when cortical neurons pretreated with 10 μmol/L nicotine for 2 h were cultured with 0.1 μmol/L colchicine for 24 h, the rate of apoptosis decreased from 62% to 38%. The phosphorlation of Akt473 increased significantly in a bell-shape time-dependent manner, which was respectively 1.3, 3.7, 2.4, 2.1 and 1.9 times compared with the control group (P<0.01). CONCLUSION:By activating the signal pathway of Akt473, nicotine may attenuate the apoptosis of cortical neurons induced by colchicines.