矢车菊-3-O-葡萄糖苷对β淀粉样蛋白25-35致海马神经细胞损伤的干预研究  

Protective effect of cyanidin-3-glucoside against injury induced by amyloid β protein 25-35 in primary cultured hippocampal neurons

作　　者：王舒敏 谭艳[2] 卢豪 庞伟[4] 李宜波[4] 张安仁 蒋与刚[4] Wang Shumin;Tan Yan;Lu Hao;Pang Wei;Li Yibo;Zhang Anren;Jiang Yugang(College of Health and Rehabilitation, Chengdu University of Traditional Chinese Medicine, Chengdu 610075, China)

机构地区：[1]成都中医药大学养生康复学院,成都610075 [2]成都军区总医院 [3]成都军区疾病预防控制中心 [4]军事医学科学院卫生学环境医学研究所

出　　处：《卫生研究》2018年第3期413-418,共6页Journal of Hygiene Research

基　　金：天津市应用基础与前沿技术研究计划重点项目(No.14JCZDJC36100);四川省科技厅应用基础研究项目(No.2015JY0158)

摘　　要：目的探讨矢车菊-3-O-葡萄糖苷(Cy-3G)的神经保护作用。方法原代培养乳鼠海马神经细胞,免疫荧光法检测细胞纯度。5μmol/Lβ淀粉样蛋白(Aβ)25-35处理细胞3 h,Cy-3G不同方式干预后,噻唑蓝(MTT)法检测细胞存活率,乳酸脱氢酶(LDH)法检测细胞上清液中LDH漏出率,DCFH-DA探针检测细胞内活性氧(ROS)含量,罗丹明123试剂盒检测细胞线粒体膜电位。结果与对照组比,5μmol/L Aβ25-35组细胞存活率显著降低(P<0.05),而40μmol/L Cy-3G共孵育3 h可有效提高细胞存活率,还可显著降低细胞上清液中LDH漏出率、升高细胞线粒体膜电位、降低细胞ROS水平(P<0.05)。结论 40μmol/L Cy-3G共孵育3 h对Aβ25-35损伤的原代海马神经细胞有保护作用。Objective To investigate the neuroprotective effect of cyanidin-3-Oglucoside（ Cy-3G）. Methods Hippocampal neurons were isolated from newborn rat and divided into different groups.（ 1） To estimate the neuroprotective effects,the cells were treated with various concentrations of Cy-3G（ 10,20,40,60,80 and 100 μmol/L） prior to（ 3 h） amyloid β protein（ Aβ） 25-35 treatment. Neuronal viability in terms of mitochondrial activity was evaluated with the colorimetric tetrazolium salt（ MTT） assay.（ 2） To explore the related mechanism,the cells were divided into normal control group,Aβ25-35 group,Aβ25-35 ＋ Cy-3G co-incubation group and Cy-3G group,then the cell viability was analyzed by Lactate dehydrogenase（ LDH） kit. The mitochondrial membrane potential and the intracellular reactive oxygen species（ ROS） were respectively detected by Rhodamine 123 and DCFH-DA probe and measured by fluorescence spectrophotometry.Results（ 1） Relatively,40 μmol/L Cy-3G and 5 μmol/L Aβ25-35 co-incubation for 3 h showed the best protection effect.（ 2） Compared with the control group,the LDH activity in cell media increased significantly in Aβ25-35 group（ P〈0. 05）,while Cy-3G co-incubation was able to reduce LDH activity in cell media significantly（ P〈0. 05）.Compared with the control group,the mitochondrial membrane potential of the Aβ25-35 group decreased significantly（ P〈0. 05）,Cy-3 G co-incubation could significantly inhibit the decrease of mitochondrial membrane potential of hippocampal neurons induced by Aβ25-35（ P〈0. 05）. Compared with the control group,the levels of intracellular ROS in the Aβ25-35 group increased significantly（ P〈0. 05）,while Cy-3G co-incubation could significantly inhibit the increase of intracellular ROS levels（ P〈0. 05）. Conclusion The right amount of Cy-3G can protect the primary hippocampal neurons from injury induced by Aβ25-35.

关 键 词：矢车菊-3-O-葡萄糖苷 海马神经细胞 Β淀粉样蛋白25-35 活性氧 

分 类 号：Q946.83[生物学—植物学] Q593.2