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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 observe the effects of novel salidroside (Sal) analogues such as 4-methoxy-2-acetamido-2- deoxy-β-D-pyranoside (MADP) on neurons damaged by glutamate in hippocampal neurons. Methods: Primary cultured rat embryonic hippocampal neurons were incubated with MADP at a concentration of 60,120,240 μmol / L or Sal at 240 μmol / L for 24 h. 125 μmol / L glutamate was used to injure hippocampal neurons for 15 min. The morphological changes of hippocampal neurons were observed by phase-contrast microscopy. The viability of hippocampal neurons was determined by methylthiazolyl tetrazolium salt colorimetry (MTT) colorimetric assay. Methyl thymol blue (MTB) Free calcium concentration. The expression of apoptosis-related genes Bcl-2, Bax and Caspase-3 were detected by real-time fluorescence quantitative PCR. Results: Pretreatment with MADP or Sal for 24 h improved the swelling and protrusion of neuronal somatic cells after glutamate-induced injury, inhibited the increase of intracellular free calcium concentration, increased the expression of Bcl-2 / Bax mRNA and decreased the expression of Caspase- 3 mRNA expression levels. MADP protects hippocampal neurons of rats better than Sal. CONCLUSIONS: Compared with Sal, MADP exerts a better antagonistic effect of glutamate on hippocampal neuron injury. The mechanism may be related to inhibition of influx of calcium, upregulation of Bcl-2 / Bax mRNA and down-regulation of Caspase-3 mRNA Horizontal correlation.