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一种定量评价离体海马脑片缺糖/缺氧损伤的新方法

Novel quantitative method for evaluating oxygen/glucose deprivation-induced injury of hippocampal slices

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【作者】 严乐勤魏尔清胡海涛张纬萍王梦令沈建中

【Author】 YAN Le qin, WEI Er qing, HU Hai Tao, et al (College of Medical Sciences, Zhejiang University, Hangzhou 310031, China)

【机构】 浙江大学医学院浙江大学医学院 浙江杭州310031浙江杭州310031浙江杭州310031

【摘要】 目的 :建立一种定量评价离体脑片缺糖 /缺氧损伤的简便、快速、灵敏的方法。方法 :离体海马脑片缺糖 /缺氧不同时间后以 2 % 2 ,3,5 -三苯基氯化四氮唑 (TTC)染色 ,用乙醇∶二甲亚砜 (5 0∶ 5 0 )抽提 TTC红色结晶产物 ,分光光度法测定 OD490 值 ,并与 LDH释放率进行比较。结果 :随着缺糖 /缺氧时间的延长 ,TTC染色 (OD490 值 )逐渐下降 ,而 LDH释放率逐渐上升 ,两结果呈负相关 (r=- 0 .933,P<0 .0 1) ,提示脑片缺血性损伤。尼莫地平、地塞米松和氯胺酮可减轻脑片缺血性损伤 ,但 ONO- 10 78无效。结论 :TTC染色法简单易行 ,且客观灵敏 ,可用于离体脑片损伤的评价及药物初筛。

【Abstract】 Objective: To establish a simple, sensitive in vitro method to evaluate oxygen/glucose deprivation (OGD) induced injury of brain hippocampal slices in rats. Methods: Rat hippocampal slices were incubated in 2% 2,3,5 triphenyltetrazolium chloride (TTC) solution after oxygen/glucose deprivation. They were then soaked in a measured volume of ethanol and dimethylsulfoxide(50∶50) to extract the TTC formazan product which was then measured by spectrophotometry. OGD induced LDH release was simultaneously measured. Results: Progressive prolongation of OGD induced hippocampal injury resulted in decreased formazan coloration as determined by spectrophotometry. There was a parallel increase in LDH release, thus a negative correlation in these two products was noted. ( r =-0.933, P <0.01). The injury was attenuated in the brain slices pre treated with nimodipine, dexa methasone, and ketamine, but not ONO 1078. Conclusion: Solvent extraction and spectrophotometric quantification of formazan represents an objective measurement of OGD induced injury of rat hippocampal slices.

【基金】 国家自然科学基金 ((39770 2 70 );浙江省自然科学基金 (3990 90 )资助项目
  • 【文献出处】 浙江大学学报(医学版) ,Journal of Zhejiang University(Medicine Edition) , 编辑部邮箱 ,2002年02期
  • 【分类号】R741.02
  • 【被引频次】17
  • 【下载频次】117
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