中文摘要：

目的研究ATP敏感性钾离子通道开放剂二氮嗪、环孢菌素A及两者协同对β-淀粉样蛋白（Aβ1-42）致原代培养基底前脑胆碱能神经元凋亡的影响。方法采用2μmol／L的鹏。对原代培养大鼠胆碱能神经元进行干预，建立阿尔茨海默病（AD）细胞凋亡模型；采用500μmol／L二氮嗪（预处理1h）、20μmol／L环孢菌素A以及两者协同对其干预，作用不同时间（24、72h）后置于倒置显微镜下观察细胞形态；采用噻唑蓝（MTT）比色法检测细胞存活率，Western blot法检测细胞内凋亡相关蛋白表达水平的变化。结果①邶142作用胆碱能神经元72h后，Bcl-2表达量降低（P〈0．01），细胞色素C、Caspase-3、Cleavedcaspase-3的表达量增加（P〈0．01，P〈0．001，P〈0．01），Bax未见明显变化，Bcl-2／Bax比值降低（P〈0．01），细胞形态发生明显损伤性变化，细胞存活率明显降低（P〈0．001）；②二氮嗪、环孢菌素A以及两者协同作用24h后Bcl-2的表达增加（P〈0．05，P〈0．01，P〈0．01），作用72h能明显增加Bcl-2的表达（P〈0．05，P〈0．01，P〈0．01），并提升Bc-2／Bax比率而抑制细胞色素C、Caspase-3、Cleavedcaspase-3表达量（P〈0．05，P〈0．01，P〈0．001），细胞存活率均增高（P〈0．01，P〈0．001）。结论Aβ1-42作用胆碱能神经元72h能促使细胞凋亡，二氮嗪与环孢菌素A及两者协同可以明显桔抗Aβ1-42致细胞凋亡作用。

英文摘要：

Objective To investigate the effects of diazoxide （ATP-sensitive potassium channel opener） and cyclosporine A and their combination on apoptosis induced by beta-Amyloid protein （Aβ1-42 ） on cultured primitive rat basal forebmin cholinergic neurons. Methods The cell apoptosis model was induced by Aβ1-42 （2μmol/L）, and then 500/μmol/L diazoxide, 20μmol/L cyclosporine A and their combination were used to interfere with it. Cell morphology was observed by an inverted microscope,changes of cell apoptosis were determined by MTT, and expressions of target proteins （ bcl-2, bax, cytochrome C, caspasce-3 and Cleaved Caspase-3） were determined by Western blot when the neurons were interfered with by the drugs for different times （24, 72h）. Results ① Being exposed to Aβ1-42 for 72h, cell activity remarkably decreased（P〈0.001）, expression of Bcl- 2 decreased （ P 〈 0.01 ） and expressions of cytochrome C, caspasce-3 and Cleaved Caspase-3 increased（ P 〈 0.01, P 〈 0.001, P〈0.01）. Expression of Bax had no change but the value of Bcl-2/Bax decreased（P 〈 0.01）. ②Expression of Bcl-2 increased when the cell apoptosis model was exposed to diazoxide, cyclosporine A and their combination for 24 h（ P 〈 0.05, P 〈 0.01, P 〈 0.01）. When the cell apoptosis model was exposed to diazoxide, cyclosporine A and their combination for 72 h, expression of Bcl-2 obviously increased （ P 〈 0.05, P 〈 0.01, P 〈 0. 001 ）, expressions of Cytochrome C, Caspasce-3 and Cleaved Caspase-3 decreased（ P 〈 0.05, P 〈 0.01, P 〈 0.001 ）, and cell activity increased（ P 〈 0.01, P 〈 0. 001 ）. Conclu- sion Being exposed to Aβ1-42 for 72 h, neuron apoptosis occurs. Diazoxide and cyclosporine A and their combination could pro- tect neurons from apoptosis induced by Aβ1-42.