川芎嗪对谷氨酸损伤海马神经元的保护作用.doc

文档介绍：川芎嗪对谷氨酸损伤海马神经元的保护作用
【摘要】目的观察川芎嗪对谷氨酸损伤海马神经元的保护作用。方法原代培养大鼠海马神经元,培养基中加入1mmol/L的盐酸川芎嗪溶液共同孵育12h后,加入1mmol/L谷氨酸损伤海马神经元20min。采用MTT法检测细胞活性;测定培养液中乳酸脱氢酶(LDH )活力的变化;细胞免疫组化检测各组细胞表达AchE。结果川芎嗪可提高谷氨酸损伤后海马神经元的存活率,抑制因谷氨酸损伤引起的细胞内LDH的释放,可促进谷氨酸损伤细胞中AChE的表达。结论川芎嗪对谷氨酸诱导海马神经元损伤有保护作用。
【关键词】川芎嗪;谷氨酸;海马神经元
: Objective To observe the protective effects of ligustrazine on glutamateinduced injury in cultured hippocampal neurons. Methods Primarily cultured hippocampal neurons from fetal rats mol/L) for 12 hours, then glutamate (1mmol/L) inutes to induce injury. Cell viability ined by biochemical method. Finally, the expression of AchE in the cultured neurons muocytochemistry. Results Ligustrazine improved the survival rate of hippocampal neurons injured by glutamate, inhibited the release of LDH from the kytoplasm injured by glutamate. The expression of AchE in the injured neurons oted by pretreatment of ligustrazine. Conclusion Ligustrazine can significantly exert protective effects on the hippocampal neuron injury induced by glutamate.
KEY TT)、二甲亚砜(DMSO) 、L多聚赖氨酸、阿糖胞苷为美国Sigma公司产品;胰蛋白酶、DMEM培养基为美国Gibco公司产品;胎牛血清为杭州四季青有限公司产品;6孔板/Ⅱ96孔板为丹麦Costar公司产品;乳酸脱氢酶(LDH)检测试剂盒为南京建成生物工程研究所产品;其他试剂均为国产分析纯。Olympus倒置相差显微镜(日本),CO2培养箱(NAPCO公司),酶联免疫检测仪(美国BIOTek公司),兔抗大鼠AchE抗体(1∶1000)、羊抗兔IgG(1∶200)、ABC及DAB显色试剂盒(美国VECTOR公司)。
海马神经元的原代培养[2] 健康孕15d的SD大鼠,麻醉后无菌条件下取胎鼠脑,解剖显微镜下取出的双侧海马,去除脑膜和血管,℃消化10min,用完全培养基(90% DMEM+10%