离体海马脑片胞外记录和HPLC检测在缺氧脑损伤研究中的应用

【作者】 赵晖；

【导师】 郑筱祥；

【作者基本信息】 浙江大学 ， 生物医学工程， 2003， 硕士

【摘要】 脑缺血疾病是威胁人类健康的主要疾病之一，它具有发病率高，死亡率高的特点，严重危害人类健康。如何防治脑缺血所致的脑组织损伤，已成为当今医学领域的重要研究课题。 本研究在离体海马脑片缺氧模型上，建立脑片胞外记录技术与高效液相检测技术相结合的方法，应用于脑缺血损伤研究中，并在此基础上考察钙调蛋白(Calmodulin，CaM)抑制剂三氟拉嗪(TFP)对脑缺氧的神经保护作用。 采用神经电生理技术，研究了海马脑片缺氧后的群峰电位(PS)的变化和TFP对其的影响，类似结果未见报道。结果发现TFP可以推迟PS的消失时间、降低缺氧损伤电位(HIP)的出现率，提高复氧后PS的恢复率和恢复程度，表明TFP对缺氧损伤的脑片有明显的保护作用。 应用高效液相色谱(High potential liquid chromatogram，HPLC)技术，研究缺氧导致的脑片孵育液中兴奋性神经递质天冬氨酸(Asp)、谷氨酸(Glu)与抑制性神经递质牛磺酸(Tau)、p-氨基丁酸(GABA)的含量变化以及TFP对这些氨基酸变化的影响。结果表明，TFP可显著地抑制缺氧所致脑片孵育液中兴奋性氨基酸含量的增加。 本研究所运用的海马脑片胞外记录技术和高效液相检测技术相联合的方法，对其他抗脑缺血药物的药理作用研究具有一定的指导意义。同时通过研究发现，TFP对脑缺氧损伤有明显的保护作用，其作用机理可能与抑制海马组织兴奋性氨基酸的释放有关。更多还原

【Abstract】 Ischemic stroke is a leading cause of death and serious, long-term disability in adults. Now researchers are studying the mechanisms of the disease and ways to prevent this injury to the brain.Using rat hippocampal slices ischemia model in vitro, We set up the method of extracellular recording technique and amino acids detection with the HPLC system to study brain hypoxic injury. With the method , We studied the neuroprotective effects of Calmodulin(CaM) antagonist trifluoperation(TFP) against hypoxic injury.The changes of PS after hypoxia and the protective effects of TFP were studied by the extracellular recording technique in rat hippocampal slices. There is no report to show that this sort of experiment has ever been done elsewhere. The results showed that treated with TFP, the disappearance time of PS after hypoxia was significantly suspended, the appearance of hypoxic injury potential (HIP) was lower and the recovery amplitude of PS after reoxygenation was significantly higher than that of OGD group. The results showed that TFP can protect brain slices from hypoxic injury.With the method of amino acids detection in the culture medium of brain slices with the HPLC system. We studied the effect of TFP on the changes of level of amino acids (Asp, Glu, Tau, GABA) in the culture medium of brain slices during OGD. It was found that OGD increase the release of amino acids and TFP can markedly inhibit the increase of the excitatory amino acids (EAAs).The extracellular recording technique in rat hippocampal slices , associated with the HPLC fluoromatric method would be helpful for further studies in pathological mechanisms to ischemic stroke.The study also showed that TFP can protect brain from hypoxic injury . The mechanism may be that TFP can reduce the increase of excitatory amino acids and protect synapyic function from hypoxic injury.更多还原