DIDS对SIN-1诱导大鼠海马神经元凋亡及PARP-1/AIF表达的影响

摘要

Aim To observe the proteetive ellects of DIDS, one kind of chloride channel Mockers, on hippocampal neuronal damage induced by NO in cukure. Methods The cultures were randomly assigned into conLrol group, 3-morpholinosydnonimine ( SIN-1 0.5 mmol o L-1 for 18 h) treatment group, SIN-1 + 4,4'-disothiocyanatostilbene-2, 2' disulfonic acid ( DIDS, 0. 1 mmol o L-1 ) group. The neuronal viabilkies were detected with the methods of MTT and the neuronal ap-optosis was assyed by Hoechst 33258 stainning and ac-tivated caspase-3 by Western blot, the both expressions of PARP-1 and A1F were also assayed with immunoflu-orscent staining. Results There was a significant protective effect of DIDS on neuronal damage in a dose-dependent manner. DIDS inhibited the activation of Caspase-3 and the expression of PARP-1/A1F. Conclusions Chloride channel activities may be involved in neuronal injury induced by NO, and its mechanism underlying the protection of chloride channel blocker a-gainst neuronal injury may relate to the down regulation of PARP-1/A1F expression.%目的 观察氯通道阻断剂4,4′-二异硫氰基芪-2,2′-二磺酸(DIDS)对NO诱导的大鼠离体海马神经元凋亡的保护作用.方法 离体培养12 d的SD大鼠海马神经元,随机分为正常对照组、3-吗啡斯德酮亚胺(3-morpholinosyndnomine,SIN-1)处理组、SIN-1+DIDS组.对各组神经元分别在相应的时间点用MTT法测定细胞生存率、Hoechst 33258测定凋亡百分数、免疫化学荧光分析检测凋亡信号蛋白PARP-1/AIF的变化、Western blot 分析凋亡蛋白Caspase-3的变化.结果 DIDS呈剂量依赖性地抑制SIN-1诱导的神经元损伤,减少凋亡发生数目,并能抑制损伤所引起的Caspase-3的激活、削弱PARP-1/AIF的表达.结论 氯通道可能参与了NO诱导的海马神经元损伤,氯通道阻断剂DIDS的保护作用可能与削弱PARP-1/AIF的表达有关.