Molecular mechanisms of general anesthetics on glutamate receptor channels : Identification of a site of action and examination of anesthetic behaviors in knock-out mice

全身麻醉药对谷氨酸受体通道的分子机制：作用位点的鉴定和基因敲除小鼠麻醉行为的检查

• 批准号: 10671407
• 负责人: SHIBUE Chieko
• 金额: $ 2.43万
• 依托单位: Niigata University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671407/
• 关键词: General anesthetics; Glutamate reveptor; NMDA receptor; Molecular biology; Site-directed mutagenesis; Knock-out mice; 麻薬; グルタミン酸受容体チャネル; サブユニット; 部位特異的変異導入法

Glutamate receptor channels mediate most of the fast excitatory synaptic transmission in the central nervous system. Recent studies suggest that inhibition of glutamate receptor channels is involved in mechanisms of actions of general anesthetics and sedatives. The present research examined the effects of general anesthetics on the recombinant glutamate receptor channels composed of various subunits. We also tested the importance of a certain amino acid residue of the subunit in the action of anesthetics by site-directed mutagenesis. Furthermore, to investigate a role of a certain subunit in the behavioral effects of general anesthetics in vivo, we examined anesthetic behaviors in the mice lacking the specific glutamate receptor channel subunit. We have found that butyrophenones selectively inhibit NMDA receptor channels composed of ε2 subunits. It was also found that opioid inhibits NMDA receptor channels at high concentrations which seem to be achieved in the cerebrospinal fluid after epidural or intrathecal administration of opioids, but seem to be higher than those observed in the blood during high dose opioid anesthesia for cardiac surgery. The action sites of butyrophenones and opioids partially overlapped with those of Mg^<2+> and dissociative anesthetics and mechanisms of these drugs involved the channel-block of the NMDA receptor channels. Furthermore, the anesthetic effects of ketamine measured by loss of righting reflex in mice lacking the ε1 subunit of the NMDA receptor channel were weaker than the wild-type mice, thus, anesthetic effects of ketamine might be mediated by the NMDA receptor channel composed of the ε1 subtunit. These results indicated the involvement of a specific subunit or a specific amino acid residue in the action mechanisms of anesthetics on glutamate receptor channels and in the anesthetic behavioral effects in vivo.

谷氨酸受体通道介导了中枢神经系统中的大部分快速兴奋性突触传播。最近的研究表明，抑制谷氨酸受体通道参与全身麻醉和镇静剂的作用机理。本研究检查了全身麻醉药对由各种亚基组成的重组谷氨酸受体通道的影响。我们还测试了亚基通过位置诱变的麻醉剂作用中某些氨基酸生活的重要性。此外，为了研究某个亚基在体内大麻醉的行为效应中的作用，我们检查了缺乏特定谷氨酸受体通道亚基的小鼠中的麻醉行为。我们发现丁二烯酮有选择地抑制由ε2亚基组成的NMDA受体通道。还发现，阿片类药物以高浓度抑制NMDA受体通道，在硬膜外或阿片类药物鞘内给药后似乎在脑脊髓液中实现，但似乎高于高剂量的阿片类麻醉期间血液中的血液中观察到的心脏外科手术。丁二烯酮和阿片类药物的作用位点部分与Mg^<2+>的动作重叠，这些药物的分离性麻醉和机制涉及NMDA受体通道的通道障碍。此外，通过缺乏NMDA受体通道的ε1亚基在缺乏正直反射的小鼠中测量的氯胺酮的麻醉作用比野生型小鼠弱，因此，NMDA受体通道可能由NMDA受体通道介导氯胺的麻醉效应。这些结果表明，特定亚基或特定氨基酸保留在麻醉剂在谷氨酸受体通道上的作用机理以及体内的麻醉行为效应中。

项目成果

遠藤裕 ら: "Near-infrared spectroscopyによる脳局所酸素飽和度の炭酸ガス反応について-TCDによる中大脳動脈血流速度との比較から-"麻酔. 47. 1090-1095 (1998)

Yutaka Endo 等人：“近红外光谱测量的局部脑氧饱和度的二氧化碳反应 - 与 TCD 测量的大脑中动脉血流速度的比较 -” 麻醉 47. 1090-1095 (1998)。

Yamakura T, et al.: "Direct inhibition of the N-methyl-D-aspartate receptor channel by high concentrations of opioids"Anesthesiology. 91(4). 1053-1063 (1999)

Yamakura T 等人：“高浓度阿片类药物直接抑制 N-甲基-D-天冬氨酸受体通道”麻醉学。

Yamakura T, et al.: "N-methyl-D-aspartate receptor channel block by meperidine is dependent on extracellular pH"Anesth Analg. 90(4). 928-932 (2000)

Yamakura T 等人：“哌啶的 N-甲基-D-天冬氨酸受体通道阻断取决于细胞外 pH”Anesth Analg。

福田悟 ら: "除細動器植込み時の脳機能と脳酸素代謝"J Anesth. 13(Suppl). 253 (1999)

Satoru Fukuda 等：“除颤器植入期间的脑功能和脑氧代谢”J Anesth 13（增刊）253（1999）。

渋江智栄子 ら: "プロボフォール麻酔覚醒30分後に痙攣様体動を来した1症例"麻酔. 48. 454 (1999)

Chieko Shibue 等：“从丙泊酚麻醉中醒来 30 分钟后发生抽搐身体运动的病例” 麻醉 48. 454 (1999)