不同浓度一氧化氮供体对培养海马神经元存活率影响的相关研究  

Effect of donator of nitrogen monoxide at different concentration on survival rate of cultured hippocampal neurons

作　　者：陈明[1] 孙红宇[2] 李晓文[1] 高天明[1] 

机构地区：[1]南方医科大学基础医学院神经生物学教研室,广东省广州市510515 [2]南方医科大学基础医学院生理学教研室,广东省广州市510515

出　　处：《中国临床康复》2006年第8期70-72,共3页Chinese Journal of Clinical Rehabilitation

基　　金：国家杰出青年基金(30125013);国家自然科学基金重点项目(30330240)~~

摘　　要：目的:观察不同浓度一氧化氮供体硝普钠对培养海马神经元存活率的影响以及不同类型钾通道阻断剂对硝普钠诱导的神经元死亡的保护作用。方法:实验于2004-11/2005-04在南方医科大学基础医学院神经生物学教研室实验室进行。①取新生两天以内SD大鼠海马组织做神经元分散细胞原代培养,培养第8天时,分别以不同浓度硝普钠(0.01mmol/L,0.1mmol/L,0.5mmol/L,1mmol/L)处理18h,采用MTT法观察硝普钠对神经元存活率的影响。②硝普钠(0.1mmol/L)处理前10min培养液内预先分别给予钾通道阻断剂四乙铵1mmol/L、4-氨基砒碇1mmol/L、Iberiotoxin(IBTX)100nmol/L或paxilline100μmol/L,18h后测定神经元存活率。结果:①低浓度硝普钠(0.01mmol/L)对培养海马神经元存活无影响(P>0.05),增大硝普钠浓度至0.1mmol/L,0.5mmol/L或1mmol/L可引起神经元存活率浓度依赖性地降低(P<0.001)。②硝普钠(0.1mmol/L)处理前10min给予不同类型钾通道阻断剂,广谱钾通道阻断剂1mmol/L四乙铵或A型钾通道阻断剂1mmol/L4-氨基砒碇均不能降低硝普钠诱导的神经元死亡(P>0.05);两种不同的特异性大电导钙激活钾通道阻断剂100nmol/LIBTX或10μmol/Lpaxilline均可完全保护硝普钠诱导的神经元死亡(P<0.001)。结论:一氧化氮供体硝普钠剂量依赖性地诱导培养新生大鼠海马神经元死亡,大电导钙激活钾通道阻断剂对一氧化氮诱导的神经元死亡具有完全保护作用,提示阻断大电导钙激活钾通道可能作为临床治疗缺血性脑损伤的新靶点。AIM： To observe the effects of different .concentrations of sodium nitroprusside （SNP）, which is the donator of nitrogen monoxide, on the survival rate of cultured hippocampal neurons and the protective effect of different kinds of potassium channel blockers on neuronal death induced by SNP. METHODS： The experiment was performed in the laboratory of Staff Room of Neurobiology, School of Basic Medical Sciences, Southern Medical University from November 2004 to April 2005, ①Primary hippocampal neuronal cultures were prepared from neonatal SD rats （aged within 2 days）, Neurons cultured at 8 days were treated with different concentrations of SNP （0.01 mmol/L, 0,1 mmol/L, 0,5 mmol/L, 1 mmol/L） for 18 hours, and then neuronal survivals were detected by MTT method, ②Different kinds of potassium channel blockers, such as 1 mmol/L tetraethylammonium, 1 mmol/L 4-aminopyridine, 100 nmol/L lberiotoxin （IBTX） or 100 μmol/L paxilline were added into culture medium ten minutes before the treatment of SNP （0.1 mmol/L） and neuronal survival rate were estimated 18 hours later. RESULTS： ①Low concentration of SNP （0.01 mmol/L） had no effect on neuronal survival （P 〉 0.05）, while higher concentration of SNP （0.1 mmol/L, 0,5 mmol/L or 1 mmol/L） reduced the percent of neuronal survival in a concentration-dependent manner （P 〈 0,001）.②Ten minutes pretreatment of SNP （0,1 mmol/L）, a extensive potassium channel blocker, 1 mmol/L tetraethylammonium or 1 mmol/L 4-aminopyridine showed no protective effects on neuronal death （P 〉 0.05）. Two kinds of large-conductance calcium-activated potassium channel specific blockers, IBTX （100 nmol/L） or paxilline （10 μmol/L） completely protected the neuronal death induced by SNP （P 〈 0,001）. CONCLUSION： SNP, which is the donator of nitrogen monoxide, induces the cultured hippocampal neuronal death in a concentration-dependent manner, which is protected by specific blockers of large-conductance calcium-activated potassiu

关 键 词：神经元 一氧化氮 钾通道 钙激活 钾通道阻断剂 细胞培养 浓度 供体 存活率 

分 类 号：R338[医药卫生—人体生理学]