谷胱甘肽抗谷氨酸诱导海马神经元损伤钙超载机制研究  

Effect of glutathione on intracellular Ca^(2+) level of hippocampal neurons caused by glutamate

作　　者：王晓虹[1] 孙长凯[2] 王苏平[1] 范明[3] 丁爱石[3] 吴燕[3] 马辉[2] 王禄[2] 

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]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：《中国现代应用药学》2006年第1期13-17,共5页Chinese Journal of Modern Applied Pharmacy

基　　金：国家自然科学基金项目(No.30070267);中国博士后科学基金项目(2001);辽宁省重点科技攻关项目(No.2001226005)。

摘　　要：目的观察还原型谷胱甘肽(GSH)及氧化型谷胱甘肽(GSSG)对谷氨酸(G lu)诱导海马神经元死亡影响及其作用机制。方法选用新生Wistar大鼠原代培养海马神经元Glu细胞毒性损伤模型,采用台盼蓝活细胞拒染、培养液乳酸脱氢酶(LDH)测定及TUNEL细胞凋亡原位检测等方法比较GSH及GSSG预处理3d对Glu细胞毒性损伤的影响,同时利用激光扫描共聚焦显微镜观测细胞内Ca2+浓度的变化,并与MK-801比较。结果GSH及GSSG均可减少Glu诱导神经元死亡,轻度抑制Glu诱导的神经元Ca2+内流,但不及MK-801。结论谷胱甘肽对Glu细胞毒性神经损伤有保护作用,除与通常公认抗氧化作用有关外,抑制Glu诱导的神经元Ca2+内流亦为其保护机制之一。OBJECTIVE To study the effect of reduced glutathione （ GSH ） and oxidized glutathione on hippocampal neuron death and intracellular Ca^2＋ level caused by glutamate and clarify its therapeutical mechanism. METHODS Glutamate cytotoxicity model of primary culture neurons from neonatal Wistar rat hippocampal was used to evaluate the effects of GSH and GSSG on neuron damage caused by Glu and compared with that of MK-801. Trypan blue dye, TUNE labeling and Lactate Dehydrogenase determination were used to evaluate neuron damage. Intracellular Ca^2＋ current was determined with laser scanning confecal microscopy （LSCM）. RESULTS GSH or GSSG cultured with neurons for 3 days could reduce LDH efflux and hippecampal neuons death ratio of induced by Glu, and could inhibit Ca^2＋ influx induced by Glu. But all that could not match that of MK-801. CONCLUSION： Glutathione can protect neurons against Glu damage. Besides as intra- and extra-cellular protective antioxidant, decreasing Ca^2 ＋ influx induced by Glu is also a mechanism of Glutathione against Glu cytotoxicity.

关 键 词：兴奋毒性 神经保护 海马神经元 谷胱甘肽 激光扫描共聚焦显微镜 

分 类 号：R964[医药卫生—药理学]