TRPC离子通道参与硝普钠诱导海马神经元凋亡  

Canonical transient receptor potential ion channel is involved in sodium nitroprusside-induced apoptosis of cultured rat hippocampal neurons

作　　者：张贺[1] 王斌[1] 李文军[1] 邹飞[1] 

机构地区：[1]南方医科大学公共卫生与热带医学学院,广东广州510515

出　　处：《南方医科大学学报》2008年第5期764-766,共3页Journal of Southern Medical University

基　　金：国家重点基础研究发展计划(2006CB504100)

摘　　要：目的探讨TRPC离子通道在一氧化氮供体硝普钠(SNP)诱导原代培养海马神经元凋亡中的作用。方法原代培养的海马神经元作为空白对照组,实验组分为3组,分别为原代培养神经元中加入SNP;SNP+TRPC通道阻断剂组;SNP+TRPC通道激活剂组。细胞处理24h后MTT比色法检测各组细胞存活率,Hoechest33342荧光染色观察细胞凋亡。结果SNP处理24h,神经元存活率为67.4%;与空白对照组有显著差异(P<0.05),SNP+TRPC阻断剂组神经元存活率分别为102%﹑92.7%,与SNP组有显著差异(P<0.05);而SNP+TRPC激活剂组神经元存活率为56.9%,与SNP组有统计学差异(P<0.05);单纯给予TRPC阻断剂或激动剂对神经元存活率无明显影响。荧光显微镜下观察,空白对照组神经元胞核均匀蓝染;SNP处理组胞核明显固缩、凝聚,可见凋亡小体;SNP+TRPC激动剂组可见大量凋亡细胞;而SNP+TRPC阻断剂组胞核均匀蓝染。结论TRPC离子通道参与SNP诱导海马神经元凋亡。Objective To understand the role of canonical transient receptor potential （TRPC） channel in sodium nitroprusside （SNP）-induced apoptosis of primary cultured rat hippocampal neurons. Methods Cultured rat hippocampal neurons were treated for 24 h with SNP, SNP＋TRPC channel blockers, or SNP＋TRPC channel activator. The viability of the neurons was determined by MTT assay, and the morphological changes of the cell nuclei were observed by Hoechst 33342 staining. Results The survival rate of neurons treated with SNP for 24 h （67.4%） was significantly decreased as compared with the untreated neurons （P〈0.01, n=8）. TRPC channel blockers （2APB and SKF96365） significantly increased the survival rate of the SNP-treated neurons （102% and 92.7%, respectively, P〈0.01）, while TRPC channel activator （OAG） significantly lowered the survival rate of the SNP-treated neurons （56.9%, P〈0.05）. Treatment of the cultured rat hippocampal neurons with the TRPC Channel blocker or activator alone did not obviously affect the survival of the cells. Under fluorescence microscope, the nuclei of the cultured rat hippocampal neurons were uniformly blue-stained.. The neurons treated with SNP exhibited such apoptotic morphological features as cell shrinkage, condensation, aggregation of nuclear chromatin and presence of bright blue,stained apoptotic body. Treatment of the cells with both SNP and OAG resulted in increased number of apoptotic neurons. Conclusion TRPC channel is involved in SNP-induced apoptosis of cultured rat hippocampal neurons.

关 键 词：硝普钠 TRPC 海马神经元 神经元凋亡 

分 类 号：R338[医药卫生—人体生理学]