NPPB对低渗诱导海马神经元凋亡的拮抗作用

目的观察体积调节性氯离子通道阻滞剂NPPB对低渗条件下神经元凋亡的影响。方法采用原代培养的大鼠海马神经元细胞,实验分为对照组(无低渗溶液处理)、低渗组、低渗加NPPB干预组;在不同时间点(10min、30min、60min),应用AnnexinⅤ/PI流式细胞检测以及免疫细胞荧光三标法(TUNEL/MAP-2/DAPI)检测各实验组神经元凋亡的情况。结果与对照组相比,神经元在低渗干预10min后凋亡细胞明显增多,60min时神经元的凋亡达到高峰(P0.01),而NPPB干预组在各时间点凋亡神经元相对低渗组明显下降(P0.01),较正常对照组增高。结论低渗条件可诱导体外培养海马神经元细胞凋亡,体积调节性氯离子通道阻滞剂NPPB可以显著降低低渗诱导的神经元凋亡,提示VRAC通道参与了低渗诱导的神经元凋亡。

ANTAGONIST EFFECT OF NPPB ON HIPPOCAMPAL NEURONS APOPTOSIS INDUCED BY HYPOTONIC SOLUTION IN VITRO

Objective To observe the effect of the volume-regulated anion channel blocker（NPPB） on hippocampal neuron apoptosis induced by the hypotonic solution in vitro.Methods Primarily cultured rat hippocampal neurons were randomly divided into three groups： control group,hypotony group and NPPB group（treated with hypotonic solution and NPPB）.At different time points（10min,30min and 60min）,the changes of neuronal apopotosis were detected by AnnexinⅤ/PI flow cytometry and immunofluorescent staining（TUNEL/MAP-2/DAPI）.Results Compared with that of the control group,the percentage of apoptotic neurons increased at 10min following hypotonic treatment and reached peak level at 60min（P〈0.01）.However,the percentage of apoptotic neurons in the NPPB groups was significantly less（（P〈0.01） than in the hypotony groups and higher（P〈0.01） than in the control groups at different time points.Conclusion The hypotonic solution can induce apoptosis of hippocampal neurons in vitro.The volume-regulated chloric anion channel blocker NPPB can remarkably inhibit neuronal apoptosis induced by hypotonic treatment.Our results suggest that the volume-regulated anion channel is involved in neuronal apoptosis induced by hypotonic treatment.