《中国康复理论与实践》

《中国康复理论与实践》 ›› 2004, Vol. 10 ›› Issue (12): 751-753.

• 基础研究 • 上一篇    下一篇

二苯乙烯苷对谷氨酸致原代培养大鼠海马神经元损伤的保护作用

李雅莉 ; 赵玲 ; 徐艳玲 ; 李林   

  1. 1.首都医科大学宣武医院药理研究室 北京市 100053;2.北京老年病医疗研究中心 北京市 100053
  • 收稿日期:2004-06-09 出版日期:2004-12-25 发布日期:2004-12-25

Protective effect of tetrahydroxystilbene glucoside on hippocampal neurons damage induced by glutamate in rats

LI Ya-li, ZHAO Ling, XU Yan-ling, et al   

  1. Department of Pharmacology, Xuanwu Hospital of Capital University of Medical Sciences, Beijing 100053, China
  • Received:2004-06-09 Published:2004-12-25 Online:2004-12-25

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摘要： 目的观察二苯乙烯苷对谷氨酸(Glu)致原代培养大鼠海马神经元损伤的保护作用。方法原代培养大鼠海马神经元细胞与不同浓度的二苯乙烯苷(5—100μmol/L)共同孵育24h,加入工具药Glu(终浓度为500μmol/L)孵育。四甲基偶氮唑盐(MTT)法测定细胞存活率;乳酸脱氢酶(LDH)漏出率法测定细胞膜损伤;使用荧光指示试剂Fluo-3/AM负载后,在激光共聚焦显微镜下观察不同细胞内Ca2+的荧光强度。结果不同浓度的二苯乙烯苷与细胞孵育24h后可明显拮抗Glu介导的神经毒性作用,细胞存活率明显增加,LDH漏出减少,细胞死亡率降低,并呈明显剂量依赖关系;细胞内的Ca2+荧光强度降低。结论二苯乙烯苷拮抗Glu诱导的神经毒作用的机制可能为选择性抑制大剂量Glu引起的Ca2+ 浓度异常升高。

关键词: 二苯乙烯苷, 谷氨酸(Glu), 海马神经元, 激光共聚焦显微镜

Abstract: ObjectiveTo observe the protective effect of tetrahydroxystilbene glucoside (TSG) on rats' hippocampal neuronal damage induced by glutamate (Glu) in the culture.MethodsHippocampus was isolated from newborn SD rats and dispersedly cultured in the medium for 9 days. Neurons were incubated with TSG (5—100μmol/L) for 24h, the cells were washed twice with Lock's solution without Mg2+,then Glu 500 μmol/L was added. Thirty min later, the reaction was terminated by washing the monolayer cells twice with the Lock's solution and then cultures were kept at 37℃ for 24h. Cell viability was measured by MTT method and cell membrane damage was determined by LDH leakage; with Fluo-3/AM as an intracellular calcium indicator and added into the bathing medium, fluorescent intensity of intracellular free calcium were observed through laser scanning confocal microscopy (LSCM).ResultsAfter the treatment with 5—100μmol/L TSG for 24h, the decrease of cell viability and the increase of LDH leakage caused by Glu was obviously resisted dose dependently. TSG inhibited increase of Ca2+ in cytoplasm, compared with model group.ConclusionTSG can significantly promote the cell viability and reduce the cell membrane damage in Glu treating hippocampal neurons. The neuroprotective activities of TGS is mediated by inhibiting Ca2+ overload in cytoplasm.

Key words: tetrahydroxystilbene glucoside, glutamate (Glu), hippocampal neuron, laser scanning confocal microscopy