红景天苷类似物MADP对谷氨酸诱导的大鼠海马神经元凋亡的保护作用  

Protective effects of a saliroside analog,MADP on rat hippocampal neurons against glutamate induced cell apoptosis

作　　者：赵婧[1] 郑艳红[1] 黄军[2] 武冰馨[2] 孙诚[1] 杨宇民[1] 

机构地区：[1]南通大学神经再生重点实验室,南通226001 [2]南通大学医学院,南通226001

出　　处：《神经解剖学杂志》2013年第6期631-636,共6页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(31271260);江苏省自然科学基金(BK2011132)

摘　　要：目的:观察新型红景天苷(salidroside,Sal)类似物4-甲氧基苯甲基-2-乙酰氨基2-脱氧-β-D-吡喃糖苷(4-methoxy-2-acetamido-2-deoxy-β-D-pyranoside,MADP)对谷氨酸(glutamate)损伤海马神经元的保护作用。方法:原代培养大鼠胚胎海马神经元,与浓度分别为60、120、240μmol/L的MADP或240μmol/L的Sal共同孵育24 h,加入125μmol/L谷氨酸损伤海马神经元15 min。使用相差显微镜观察海马神经元的形态变化;四甲基偶氮唑盐(methyl thiazolyl tetrazolium salt colorimetry,MTT)比色法测定细胞活力;甲基百里香酚蓝(methyl thymol blue,MTB)法测定细胞内游离的钙离子浓度;采用实时荧光定量PCR技术检测细胞凋亡相关基因Bcl-2,Bax,Caspase-3的表达水平。结果:MADP或Sal预处理24 h可改善谷氨酸损伤后神经元胞体的肿胀和突起的淡化,抑制胞内游离钙离子浓度的上升,提升Bcl-2/Bax mRNA的表达水平,降低Caspase-3 mRNA的表达水平。MADP对大鼠海马神经元的保护作用优于Sal。结论:与Sal相比较,MADP更好的发挥了拮抗谷氨酸对海马神经元损伤的作用,其机制可能与抑制钙离子内流,上调Bcl-2/Bax mRNA及下调Caspase-3 mRNA的表达水平相关。Objective： To investigate the protective effects of a novel analog of salidroside （Sal）, 4-methoxy benzyl-2- acetamido-2-deoxy-β-D-pyranosides （MADP）, on glutamate induced cell injury in cultured hippocampal neurons. Methods： Primarily cultured rat embryonic hippocampal neurons were incubated with different concentrations of MADP （60, 120,240 μmol/L, respectively） or Sal （240μmol/L） for 24 hours, and then 125 μmol/L glutamate was added for 15 min to induce apoptosis. Phase contrast microscope was used to observe cell morphological changes. Cell viability was assessed by methyl thiazolyl tetrazolium salt color imetry（MTr） assay; the concentration of intracellular free calcium ion was measured by the method of methyl thymol blue（MTB） ; the mRNA levels of Bcl-2, Bax and caspase-3 were tested by real time quantitative PCR （Q-PCR）. Results： 24 h-pretreatment with MADP or Sal attenuated glutamate-induced cell apoptosis, as evidenced by the cell morphology, decreased intracellular free calcium ion concentration, up-regulated Bcl- 2/Bax mRNAs and down-regulated caspase-3 mRNA. Moreover, the neuroprotective effects of MADP were superior to those of Sal. Conclusion： MADP has more potent neuroprotective effects as compared to Sal, which was probably related to its abilities to reduce calcium ion overload in cytoplasm, to up-regulate Bcl-2/Bax mRNA expression and down-regulateCaspase-3 mRNA expression.

关 键 词：红景天苷类似物 海马神经元 谷氨酸 细胞凋亡 神经保护 大鼠 

分 类 号：R285.5[医药卫生—中药学]