钙通道在SIN-1诱导大鼠海马神经元凋亡中的作用

摘要 背景 氯通道参与SIN-1引起的大鼠海马神经元凋亡过程已有报道，而钙通道在这种类型的凋亡中的作用目前未见报道。目的 探讨钙通道在SIN-1诱导大鼠海马神经元凋亡中的作用。方法 方法 离体培养12天的SD大鼠海马神经元，随机分为正常对照组、3-吗啉斯德酮胺(SIN-1)处理组: (SIN-1 1.0 mmol •L-1）、SIN-1处理后加4-4-二异硫氰基苯-2,2’-二磺酸（DIDS，0.1 mmol •L-1）、4-乙酰氨基-4’-异氰酸芪-2,2’-二磺酸(SITS, 1.0 mmol •L-1)、SIN-1处理后加氯化镉（CdCl2）、SIN-1处理后加SITS和CdCl2及SIN-1处理后加DIDS和CdCl2。药物作用时间为18h。DNA荧光染色观察神经元形态及检测凋亡数目的变化，MTT法检测神经元的生存率。结果 SIN-1可以诱导42.32±1.34%的神经元死亡，与正常对照组相比有显著差异（P<0.05）,SIN-1处理的神经元若用氯通道阻断剂SITS或DIDS，生存率分别为87.33±2.03%和83.23±1.01%。SIN-1处理的神经元单独用CdCl2或CdCl2与SITS或DIDS合并使用时，细胞生存率分别为：66.39±2.09%、88.15±1.13%和83.16±2.00%。DNA荧光染色显示，SIN-1处理的神经元若用氯通道阻断剂可以明显减少凋亡百分数，而钙通道阻断剂CdCl2没有明显的抗凋亡作用。结论 氯通道可能参与了SIN-1诱导的大鼠海马神经元凋亡，而钙通道的作用不大。

Effects of calcium channel on 3- morpholinosydnonimine-induced rat hippocampal neuronal apoptosis

Previous studies have demonstrated that increased chloride channel activity plays a role in nitric oxide-induced neuronal apoptosis in the rat hippocampus. The present study investigated the effects of the broad-spectrum calcium channel blocker CdCl2 on survival rate, percentage of apoptosis, and morphological changes in hippocampal neurons cultured in vitro, as well as the effects of calcium channels on neuronal apoptosis. The chloride channel blockers 4-acetamido-4'-isothiocyanatostilbene-2, 2'-disulfonic acid (SITS) or 4, 4'-diisothiocyanostilbene-2, 2'-disulfonic acid (DIDS) increased the survival rate of 3-morpholinosydnonimine (SIN-1)-treated neurons and suppressed SIN-1-induced neuronal apoptosis. The calcium channel blocker CdCl2 did not increase the survival rate of neurons and did not affect SIN-1-induced apoptosis or SITS- or DIDS-suppressed neuronal apoptosis. Results demonstrated that calcium channels did not significantly affect neuronal apoptosis.