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红景天苷对谷氨酸损伤海马神经元的保护作用

PROTECTIVE EFFECTS OF SALIDROSIDE ON GLUTAMATE INDUCED NEUROTOXICITY IN CULTURED HIPPOCAMPAL NEURONS

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【作者】 陈霞周松林顾晓松丁斐

【Author】 CHEN Xia~1,2,3,ZHOU Song-lin~2,GU Xiao-song~2,DING Fei~2(1.School of Medicine,Soozhow University,Suzhou 215123;2.The Jiangsu Key Laboratory of Neuroregeneration,Nantong University,Nantong 226001;3.Department of Pharmacology,Nantong University,Nantong 226001,China)

【机构】 苏州大学医学院南通大学江苏省神经再生重点实验室南通大学江苏省神经再生重点实验室 苏州215123南通226001南通大学医学院药理学教研室

【摘要】 目的观察红景天苷(salidroside)对谷氨酸(Glu)损伤海马神经元的保护作用。方法胚鼠原代培养海马神经元,与不同浓度的红景天苷(10、20和40mg/L)共同孵育24h,加入125μmol/L谷氨酸损伤海马神经元15min。采用MTT法检测细胞活性;生化法测定培养液中乳酸脱氢酶(LDH)活力的变化;膜联蛋白-V-FITC(annexin V-FITC)和碘化吡啶(PI)双标染色以及Hoechst染色观察细胞凋亡情况;激光共焦显微镜观察细胞内Ca2+的变化。结果红景天苷可明显改善谷氨酸损伤后细胞的活性,抑制谷氨酸损伤后培养液中LDH的活力,降低谷氨酸引起的细胞凋亡率,降低谷氨酸损伤后细胞内的Ca2+浓度。结论红景天苷可明显拮抗谷氨酸损伤海马神经元的作用,其机制可能与抑制谷氨酸引起的Ca2+内流有关。

【Abstract】 Objective To observe the protective effects of salidroside on glutamate-induced injury in cultured hippocampal neurons.Methods Primarily cultured hippocampal neurons from fetal Wistar rat were incubated with salidroside(10,20 and 40mg/L) for 24 hours,then glutamate(125μmol/L) was added for 15 minutes to induce injury.Cell viability was detected by MTT assay,the vigor of LDH was determined by biochemistry method,the apoptosis rates were anallyzed using Annexin V-FITC and PI labelling and Hoechst 33342 staining and flow cylometric assay.Fluorescent intensity of intracellular free calcium was observed with laser scanning confocal microscopy(LSCM).Results After the pretreatment with salidroside for 24 hours,the increases of LDH vigor and apoptosis rates and the decrease of cell viability caused by glutamate were resisted obviously.Salidroside inhibited the increase of Ca~2+ in cytoplasm significantly.Conclusion Salidroside can significantly resist the injury induced by glutamate.The neuroprotective activities of salidroside can be related to its ability to reduce Ca~2+ overload in cytoplasm.

【关键词】 红景天苷谷氨酸海马神经元细胞培养大鼠
【Key words】 SalidrosideGlutamateHippocampal neuronsCell cultureRat
【基金】 国家973计划子课题资助(2003CB515306)
  • 【文献出处】 解剖学报 ,Acta Anatomica Sinica , 编辑部邮箱 ,2008年03期
  • 【分类号】R285
  • 【被引频次】33
  • 【下载频次】708
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