灯盏花素含药血清对谷氨酸致原代海马神经元损伤的影响  

Protection of Serum Containing Breviscapine on Cultured Hippocampal Neuron Injury Induced by Glutamate

作　　者：王丽君[1] 郭芳[2] 

机构地区：[1]北京市第六医院药剂科,北京100007 [2]河北医科大学药理教研室,石家庄050017

出　　处：《现代生物医学进展》2010年第3期459-461,F0002,共4页Progress in Modern Biomedicine

摘　　要：目的:从细胞水平研究注射用灯盏花素对谷氨酸致大鼠原代海马神经元损伤的保护作用及其作用机制。方法:采用中药血清药理学方法,制备含药血清;原代培养大鼠乳鼠大脑海马神经元并经鉴定后,以谷氨酸复制损伤模型,以5%含药血清干预,在透射电镜下及经碘化丙啶和Hoechst33342双染后荧光显微镜下观察海马凋亡神经元的形态学变化,并进行检测:MTT法测定细胞存活率,生化法检测LDH漏出率、丙二醛(MDA)含量、细胞释放的NO量、细胞内tNOS活性和iNOS活性。结果:灯盏花素高、低剂量组均能明显增加海马神经元存活率,而LDH漏出率、丙二醛(MDA)含量、一氧化氮释放、总一氧化氮合酶活性和诱导型一氧化氮合酶活性(p<0.05,p<0.01)明显降低。结论:灯盏花素对谷氨酸致原代培养大鼠海马神经元损伤具有保护作用,其作用机制可能与其能改善能量代谢、稳定细胞膜、抗脂质过氧化、降低一氧化氮合酶的活性、减少一氧化氮释放有关。Objective： To investigate the protective effects and mechanism of Breviscapine on cultured rats hippocampal neuron injury induced by glutamate in the cell level. Methods： The drug-contained serum were obtained by the Seropharmacological method. When the primary hippocampal neurons of rat cerebral were cultured for 10 days, the mode were builded in which hippocampal neurons were exposed to 500μmol·L^-1 Glutamate acid （Glu）for 20 minutes and the 5% drug-contained serum including normal, model, 0.05 g·kg-1 Nimodipine （Nim）, 0.20 g·kg^-1 Breviscapine and 0.05g·kg^-1 Breviscapine were added to the nutrient solution of cultured neurons. In this study, the viability of cultured primary hippocampal neurons were detected by MTT assay, and the release of lactate dehydrogenase （LDH）, malonydiadehyde （MDA） content and Nitric oxide（NO）content, the activity of Nitric oxide synthase（NOS）and Induced nitric oxide synthase （iNOS）were also detected. Results： The results showed that, when the hippocampal neurons were exposed to Glu, the viability of cultured primary hippocampal neurons decreased dramatically, the release of LDH, MDA content, Nitric oxide（NO）content, the activity of Nitric oxide synthase（NOS）and Induced nitric oxide synthase（iNOS）increased obviously compared with control. The Breviscapine could decreased the release of LDH, MDA content, Nitric oxide（NO）content, the activity of Nitric oxide synthase（NOS） induced nitric oxide synthase （iNOS）. Conclusions： Breviscapine has protective effects on glutamate excitotoxicity of cultured primary hippocampal neurons of rats, and the mechanism of which may be related with improving energy metabolism, stabilizing cellular membrane, anti-liqid-peroxidation, reducing NO content, the activity of NOS and iNOS.

关 键 词：灯盏花素 谷氨酸 海马神经元 一氧化氮 一氧化氮合酶 

分 类 号：Q953[生物学—动物学] R385.5[医药卫生—医学寄生虫学]