气-液交界式浴槽的设计及其在探讨海马脑片H_2O_2损伤中的应用

Design of an Air-fluid Interface Chamber and Application for Researching Free Radical-induced Injury in Rat Hippocamcal Slices

目的 设计气 液交界式浴槽,并用于探讨海马脑片H2O2损伤的机制.方法 气 液交界式浴槽包括内室和外室.内室的主要结构为嵌入体,用于放置海马脑片.外室的主要作用是为海马脑片提供足够的氧气.另外用生化、形态学、电生理的方法来验证浴槽的可行性.最后观察H2O2对海马脑片群峰电位的影响.结果 浴槽能够长时间(1～8h)的维持海马脑片活性.H2O2能够浓度依赖性损伤海马脑片,同时NMDA受体阻断剂氯胺酮对H2O2诱导的海马脑片损伤具有明显的保护作用.结论 浴槽可以适用于生化和形态学以及电生理等研究,并发现海马脑片H2O2损伤至少部分是通过NMDA受体发挥作用的.

ObjectiveTo design an air-fluid interface chamber and its application for researching the mechanism of hydrogen peroxide-induced injury in rat hippocampal slices.MethodsThe interface chamber was made of two components: inner chamber and external chamber. Hippocampal slices were placed on the nylon net in the insert and gassed by pre-warmed oxygen. In addition, feasibility of slice reactions in the chamber was tested by biochemical, morphological and eletrophysiological methods. We also observed concentration-dependent injury by hydrogen peroxide on population spike, and effect of ketamine, an NMDA receptor antagonist, on the injury in the hippocampal slices.ResultHippocampal slices maintained active in long terms (1-8 h). Hydrogen peroxide inhibited the activity of the hippocampal slices concentration-dependently, and ketamine inhibited injury induced by hydrogen peroxide.ConclusionThe chamber developed in this study is suitable for evaluating activity of the slices, and can be used in the biochemical, morphological and eletrophysiological studies of brain slices. Hydrogen peroxide-induced injury of the hippocampal slices is, at least in part, mediated via NMDA receptor.