Development of the in vivo monitoring system of nitric oxide using a micro electrode.

使用微电极开发一氧化氮体内监测系统。

• 批准号: 07557019
• 负责人: YAMATODANI Atsushi
• 金额: $ 9.22万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557019/
• 关键词: Nitric oxide (NO); Carbon electrode; Metalloporphyrins; Cyclicvoltammetry; Nafion; Cation polymer; alpha-cyclodextrin; Endothelial cells; NO検出電極; 被覆電極; イオン性高分子; α-シクロデキストリン; in vivo測定; ナフィオン膜; 刺入型電極; in vivo計測

Following the electrochemical analysis of the oxidation reaction mechanism of nitric oxide(NO) on the electrode surface and examination of the materials for the electrode itself and its surface coating, we developed three types of electrode for the detection of trace amount of NO in biological tissues ; (1) a glassy carbon electrode covered with Nafion film containing iron porphyrins, (2) a pencil lead elecrode covered with Nafion, alpha-cyclodextrin and PDDA (cation polymer), and(3) a carbon fiber electrode covered with Nafion and PDDA.The interference in the detection of NO by several redox substances existed in tissues such as NO_2, ascorbic acid and catecholamines, could be effectively abolished by coating the electrode with Nafion, PDDA and alpha-cyclodextrin. The sensitivity for No was increased by adding iron porphyrins in Nafion film due to its catalytic properties. Thus, the electrodes could detect 10^9MNO in solution.Using the glassy carbon electrode coated with Nafion film containing iron porphyrins, we could detect NO release from the isolated rat thoracic aorta stimulated by acetyl choline, and the concentration of NO was estimated about 100nM.The specificity was confirmed by the evidence that the amperometric signal was diminished by removal of the endothel from the blood vessel or addition of L-NAME,a NO synthase inhibitor. Furthermore, the electrode could detect NO release from the cultured bovine endothelial cells. We are trying in vivo application of the detection system using the carbon fiber electrode, and the result is promising.

通过对一氧化氮（NO）在电极表面氧化反应机理的电化学分析，以及对电极本身材料和电极表面涂层材料的考察，研制了三种用于检测生物组织中痕量NO的电极;（1）覆盖有含铁卟啉的Nafion膜的玻碳电极，（2）覆盖有Nafion、α-环糊精和PDDA的铅笔芯电极用Nafion、PDDA和α-环糊精包被碳纤维电极，可以有效地消除组织中存在的几种氧化还原物质NO_2、抗坏血酸和儿茶酚胺对NO测定的干扰。在Nafion膜中加入铁卟啉，由于其催化性质，提高了对NO的灵敏度。用含有铁卟啉的Nafion膜修饰玻碳电极，我们可以检测到乙酰胆碱刺激离体大鼠胸主动脉释放的NO，NO浓度约为100 nm，去除血管内皮细胞或加入内皮细胞后，电流信号减弱，证实了其特异性。L-NAME，一氧化氮合酶抑制剂。此外，该电极可检测培养的牛内皮细胞释放的NO。我们正在尝试使用碳纤维电极的检测系统的体内应用，结果是有希望的。

项目成果

池田修、小林敬明、三宅幹夫、北島彰、大和谷厚: "一酸化チッ素の電解酸化における金属ポルフィリンの触媒作用" 電気化学協会北陸支部秋季大会講演要旨集. 9-10 (1997)

Osamu Ikeda、Takaaki Kobayashi、Mikio Miyake、Akira Kitajima、Atsushi Yamatoya：“金属卟啉在一氧化氮电解氧化中的催化作用”电化学学会北陆分会秋季会议记录 9-10 (1997)。

O.Ikeda, A.Yamatodani, M.Miyake, A.Kawaguchi: "Assay methods for the detection of nitric oxide in solution" Jpn.J.Med.Electro.Biol.Engi.33(s2). 164 (1995)

O.Ikeda、A.Yamatodani、M.Miyake、A.Kawaguchi：“溶液中一氧化氮检测的测定方法”Jpn.J.Med.Electro.Biol.Engi.33(s2)。

池田 修、小林 敬明、三宅 幹夫、北島 彰、大和谷 厚: "一酸化チッ素の電解酸化における金属ポルフィリンの触媒作用" 電気化学協会北陸支部秋季大会講演要旨集. 9-10 (1997)

Osamu Ikeda、Takaaki Kobayashi、Mikio Miyake、Akira Kitajima、Atsushi Yamatoya：“金属卟啉在一氧化氮电解氧化中的催化作用”电化学学会北陆分会秋季会议记录 9-10 (1997)。

池田 修、松村 隆、小村 照寿、望月 敬也、林 亮、三宅 幹夫、北島 彰、大和谷 厚: "生体中の一酸化チッ素検出用微小電極に関する研究" 電気化学協会北陸支部秋季大会講演要旨集. 11-12 (1997)

Osamu Ikeda、Takashi Matsumura、Teruhisa Komura、Keiya Mochizuki、Ryo Hayashi、Mikio Miyake、Akira Kitajima、Atsushi Yamatoya：“用于检测活体中一氧化氮的微电极的研究”电化学会北陆分会秋季会议摘要第 11-12 卷。 (1997)

O.Ikeda, T.Kobayashi, M.Miyake, A.Kitajima, A.Uno, A.Yamatodani: "Electrocatalytic oxidation of nitric oxide by metalloporphyrins dissolved in Nafion film." J.Electroanalytical Chem.(印刷中). (1998)

O.Ikeda、T.Kobayashi、M.Miyake、A.Kitajima、A.Uno、A.Yamatodani：“溶解在 Nafion 膜中的一氧化氮的电催化氧化”（J.Electroanalytical Chem）。 ）