目的 在离体脑片缺血性损伤模型上，建立筛选与评价神经保护药和抗炎药的方法。方法 小鼠离体脑片缺氧缺糖／再灌诱导（OGD/RP）的模型上，以2,3,5-三苯基氯化四氮唑（TTC）产物formazan定量测定评价脑片活性，观察不同药物离体给药或整体预给药7 d对脑片损伤的保护作用。结果 抗氧化剂依达拉奉、钙拮抗剂尼莫地平及神经生长因子可显著减轻脑片的缺血性损伤；半胱氨酰白三烯受体拮抗剂ONO-1078和孟鲁司特、糖皮质激素地塞米松及中枢抗炎药米诺环素无明显作用。整体给予地塞米松（0.2，2,5 mg·kg-1）能减轻脑片对缺血损伤的程度；孟鲁司特仅大剂量给药（15 mg·kg-1）有增强作用。结论 离体小鼠脑片TTC产物formazan定量分析评价药物抗缺血性损伤作用的方法，可用于初筛具有直接神经保护作用药物。
Abstract
OBJECTIVE To establish a method for screening and evaluating the effects of candidate drugs on oxygen-glucose deprivation/reperfusion (OGD/RP)-induced injury in mouse brain slices METHODS After oxygen-glucose deprivation for 15 min followed by reperfusion for 30 min, brain slices were stained with 2,3,5-trihenylterzolium chloride (TTC). TTC product formazan was measured to evaluate the viability of brain slices. The protective effects of drugs on OGD/RP injury were determined using formazan as an index. Drugs were added into the incubation media in vitro or injected i.p. for 7 d before decollation RESULTS Edaravone (an antioxidant), nimodipine (a calcium channel antagonist) and nerve growth factor (NGF) markedly attenuated OGD/RP-induced injury, but ONO-1078, montelukast (a cysteinyl leukotriene receptor antagonist)，dexamethasone (a glucocorticosteroid drug) and minocycline (a central anti-inflammatory drug) had no significant effects. Pretreatment with dexamethasone (0.2, 1, 5 mg·kg-1 per day for 7 d) significantly increased the tolerance of brain slices to OGD/RP injury, but montelukast showed this effect only at a higher dose (15 mg·kg-1) CONCLUSION Measurement of TTC product formazan is a simple and useful method for evaluating drug effects on ischemic injury in brain slices in vitro, and can be used to screen drugs that possess direct neuroprotective effects.
关键词
脑缺血 /
脑片 /
缺氧缺糖/再灌(OGD/RP) /
2 /
3 /
5-三苯基氯化四氮吐(TTC) /
神经保护剂
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Key words
cerehral ischemia /
brain slices /
oxygen-glucose deprivation/reperfusion (OGD/RP) /
2 /
3 /
5-trihenylteizolium chloride ('ITC) /
neuroprotactme agents
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脚注
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基金
国家自然科学基金资助项目（30271498）和浙江省自然科学基金资助项目（399090）
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