The Study Of Human Recombinant GABA-A Receptor Binding Sites for General Anesthec Drugs

我们采用膜片钳技术研究了不同麻醉药物对Sf 9细胞表达的重组人GABA-A受体的影响。Sf 9细胞和人类重组GABA-A受体的表达直到去年都是由九州大学医学部生理学第二部门提供的，但今年我们可以在我们的部门管理所有程序。我们研究了苯二氮卓类药物对该电流影响的机制。苯二氮卓类药物在治疗焦虑症、失眠症和癫痫症方面已确立疗效。众所周知，它们通过积极调节GABA对GABA受体的作用而产生药理作用。但细节仍不清楚。近年来，黄酮类化合物对苯并二氮杂卓类化合物的亲和力较高.它的分子结构类似于地西泮。我们使用了几种黄酮衍生物对人重组GABA-A受体的影响，但没有特别的变化occurrenced.Effects的挥发性麻醉剂，异氟烷，对甘氨酸能微型抑制性突触后电流（mIPSCs）的影响，主要是研究在机械分离的大鼠三叉神经核神经元连接完整的甘氨酸能神经元间突触前神经末梢。使用制霉菌素穿孔膜片记录配置在电压钳条件下记录mIPSC。异氟醚使酶解神经元的甘氨酸浓度-反应曲线向左平行移动，而不改变最大反应。异氟烷可逆地增加了mIPSC的频率，延长了衰减时间常数，而不影响平均振幅。在不含Ca^2+的外部溶液中也观察到异氟烷存在时mIPSC频率的增加。

We investigate the effects of various anesthetic drugs using patch-clamp technique on human recombinant GABA-A receptors expressed in Sf9 cells. Sf9 cells and expression of human recombinant GABA-A receptors were provided by the second department of Physiology, Faculty of Medicine, Kyushu University until last year, but we can manage all the procedure in our departments this year. We examined the mechanism of the effects of Benzodiazepines on this current. Benzodiazepinees have established efficacy in the treatment of anxiety, insomnia and epilepsy. And it is well known that they produce their pharmacological effects by positively modulating the action of GABA at GABA receptors. But still the detail is remained unclear. In recent Years, Flavone derivatives is reported that its affinity for benzodiaz epine is quite high. Its molecular structure is similar to diazepam. We use several fravone derivatives on human recombinant GABA-A receptors and investigate the effect, but no particular change was occurred.Effects of a volatile anesthetic, isoflurane, on glycinergic miniature inhibitory postsynaptic currents (mIPSCs) were mainly investigated in the mechanically dissociated rat trigeminal nucleus neurons attached with intact glycinergic interneuronal presynaptic nerve terminals. The nystatine-perforated patch recording configuration was used to record the mIPSCs under the voltage-clamp conditions. Isoflurane parallely shifted the glycine concentration-response curve of enzymatically dissociated neurons to the left without changing the maximum response. Isoflurane reversibly increased the frequency of the mIPSCs and prolonged the decay time constant without affecting the mean amplitude. The increase in the frequency of mIPSCs in the presence of isoflurane was also observed in a Ca^<2+>-free external solution.