N-乙酰半胱氨酸抗谷氨酸诱导海马神经元损伤的研究  

Neuron protection of N-acetylcysteine on hippocampal neuron against glutamate excitotoxicity

作　　者：王晓虹[1] 孙长凯[2] 王苏平[1] 李艳华 范明[4] 丁爱石[4] 

机构地区：[1]大连市中心医院神经内一科,辽宁大连116033 [2]大连医科大学脑疾病研究所,辽宁大连116027 [3]大连市甘井子区辛寨子地区医院,辽宁大连116033 [4]军事医学科学院基础医学研究所神经生物研究室,北京100850

出　　处：《中风与神经疾病杂志》2005年第2期141-144,i002,共5页Journal of Apoplexy and Nervous Diseases

基　　金：国家自然科学基金项目 (3 0 0 70 2 67);中国博士后科学基金项目 (2 0 0 1);辽宁省重点科技攻关项目 (2 0 0 12 2 60 0 5 )

摘　　要：目的　观察N-乙酰半胱氨酸(N- acetylcysteine,NAC)对不同浓度谷氨酸(Glutamte,Glu)诱导海马神经元损伤的影响,探讨其作用机制,评价毒性作用。方法　采用台盼蓝活细胞拒染与TUNEL法比较不同浓度NAC预处理3d给药与细胞毒性暴露后快速给药对10 0μmol/ L、5 0 0μm ol/ L Glu诱导体外培养海马神经元损伤的影响,并与MK- 80 1比较;利用激光扫描共聚焦显微镜(L SCM)观测细胞内Ca2 + 浓度变化;采用台盼蓝活细胞拒染、原子力显微镜(AFM)及细胞内酯酶活性判定方法评价NAC毒性。结果　NAC可降低10 0μmol/ L Glu诱导的海马神经元死亡率,以预处理组为佳,1m mol/ L NAC预处理保护效果类似于10μmol/ L MK - 80 1;NAC对5 0 0μm ol/ L Glu诱导的海马神经元损伤无保护作用。1mm ol/ L NAC预处理抑制Glu诱导的神经元Ca2 + 内流。经10 0 mmol/ L NAC作用的细胞虽然形态完整,但台盼蓝染色蓝染,失去对Glu毒性的反应性;AFM扫描见神经元细胞膜皱缩;培养基Ca2 +经Fluo- 3(AM)标记后L SCM下无激发荧光。结论　NAC对轻度Glu细胞毒性损伤有保护作用,预防性用药效果更优越。认为抑制Glu诱导的神经元Ca2 +内流为其保护机制之一。Objective To study the effect and mechanism of N-acetylcysteine (NAC) on hippocampal neuron death caused by glutamate at different concentration,and evaluate its toxicity.Methods Glutamate cytotoxicity model of primary culture neurons from neonatal Wistar rat hippocampal was used to evaluate the effects of different treatment of NAC on neuron damage caused by Glu and compared with that of MK-801.Trypan blue dye,Lactate Dehydrogenase determination and TUNE labeling were used to evaluate neuron damage.Trypan blue dye,Atomic Force Microscopy scan and enzyme activity observation was used to evaluate toxicity of Gluthion.Results NAC prevented neuron death induced by 100μmol/L Glu, and the effect of pre-treatment was more superior.There was no difference between the protective effects of pre-treatment group of 1mmol/L NAC and 10μmol/L MK-801 group.For 500μmol/L Glu,NAC had no protective effects.Although the configuration of cultured neurons was integrity after treated with NAC of 100mmol/L,all the cells were stained with trypan blue;membrane of neuron was crimpled under Atomic Force Microscopy scan;cultures marked with Fluo-3(AM)had no fluorescence under LSCM and neurons had no response to Glu.Conclusion NAC can protect neurons against light Glu damage,and preventive use is the most efficiency.NAC at higher concentration can fix tissue.

关 键 词：海马神经元损伤 N-乙酰半胱氨酸 谷氨酸 激光扫描共聚焦显微镜 CA^2+内流 mol/L 细胞内CA^2+ MK-801 mol/L TUNEL法 原子力显微镜 NAC 不同浓度 保护作用 Glu mol/l 台盼蓝染色 预处理 作用机制 毒性作用 细胞毒性 体外培养 

分 类 号：R338.26[医药卫生—人体生理学] R563[医药卫生—基础医学]