Analysis of motilin receptor in human gastric antrum for clinical application of motilide

人胃窦胃动素受体分析及其临床应用

• 批准号: 06454256
• 负责人: ITOH Susumu
• 金额: $ 4.67万
• 依托单位: Gunma University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06454256/
• 关键词: motilin.; motilin receptor.; erythromycin derivatives.; smooth muscle of the gastrointestinal tract.; autoradiography.; isolated smooth muscle cell.; receptor binding assay.; smooth muscle contraction

1.Since motilides are not permitted to use in human, the biological activity of motilides was studied in conscious dogs with force transducers chronicanlly implanted in the gastrointestinal tract. As a result, EM523 induced strong contractions in the fed state in addition to the fasted state. EM523-induced contractions in the stomach is mediated through cholinergic neurons including 5-HT3 receptors in the stomach. Moreover, it was also found that EM523 induced contractions in the stomach through substance P neurons without cholinergic pathways.2.It was found that EM523 stimulates pancreatic islet hormones through vagally cholinergic muscarinic receptors.EM574 and motilin stimulated contractile activity in isolated smooth muscle of human gastric antrum. EM574 also stimulated contractile activity of smooth muscle strips of human gastric antrum in a dose-dependent manner, and these contractions were TTX-resistant.In radioreceptor binding study, it was found that motilin receptors were present in the smooth muscle tissue of the human gastric antrum, and the specific binding was replaced by EM574 in a dose-dependent fashion.In the study of autoradiography, it was found that specific bindings of motilin were found in the smooth muscle layr of the human gastric antrum, and these bindings were also inhibited by addition of EM574 in a dose-dependent manner.Since the binding of motilin in the membrane fraction of the human stomach was very low, iodinated motilin was bound to the homogenate preparation of smooth muscle in the human stomach, and the bound was analyzed by means of a SDS gel electrophoresis. The three different bands in molecular weight were detected, and a single peak could be obtained from one of the bands, and now further be purifying.

1.由于胃泌肽不允许用于人体，因此在清醒的狗胃肠道中长期植入力传感器，研究胃泌肽的生物活性。结果，除了禁食状态外，EM 523还在进食状态下诱导了强烈的收缩。EM 523诱导的胃收缩通过胃中包括5-HT 3受体的胆碱能神经元介导。EM 523通过迷走神经胆碱能受体刺激胰岛激素，EM 574和胃动素刺激离体人胃窦平滑肌的收缩活动。EM 574对人胃窦平滑肌条也有剂量依赖性的收缩作用，但这种收缩是抗TTX的.在放射受体结合研究中，发现人胃窦平滑肌组织中存在胃动素受体，EM 574以剂量依赖性的方式取代胃动素受体的特异性结合.在放射自显影研究中，EM 574对胃动素受体的特异性结合具有剂量依赖性，EM 574对胃动素受体的特异性结合具有剂量依赖性.发现胃动素在人胃窦的平滑肌层中有特异性结合，并且这些结合也被加入EM 574以剂量依赖的方式抑制。由于胃动素在人胃的膜部分中的结合非常低，将碘化胃动素与人胃平滑肌的匀浆制备物结合，并通过SDS凝胶电泳分析结合。检测到三条分子量不同的谱带，其中一条谱带可得到单一峰，现进一步纯化。

项目成果

坂井貴文,他: "Biotynyl C-terminal-extended motllin as a biologically active receptor probe." Peptides. 15. 257-262 (1994)

Takafumi Sakai 等人：“Biotynyl C 末端延伸的 motllin 作为生物活性受体探针。”15. 257-262 (1994)

坂井貴文,他: "Autoradiographic study of motilin binding sitcs in the rabbit gastrointestinal tract." Regul. Pept.53. 249-257 (1994)

Takafumi Sakai 等人：“兔胃肠道中胃动素结合位点的放射自显影研究”，Pept.53（1994）。

Sasaki, T., Satoh, M., Sonobe, K.Nakajima, M., Shiba, Y.and Itoh, Z.: "Autoradiographic study of motilin binding sites in the rabbit gastrointestinal tract." Regul.Peptides. 63. 249-257 (1994)

Sasaki, T.、Satoh, M.、Sonobe, K.Nakajima, M.、Shiba, Y.和 Itoh, Z.：“兔子胃肠道中胃动素结合位点的放射自显影研究”。

坂井貴文,他: "Biotynyl C-terminal-extended motilin as a biologically active receptor probe." Peptides. 15. 257-262 (1994)

Takafumi Sakai 等人：“Biotynyl C 末端延伸胃动素作为生物活性受体探针。”15. 257-262 (1994)

柴 義博,他: "Effect of nonpeptide motilin agonist EM523 on release of gut and pancreatic hormones in conscious dogs." Gastroenterology. 110. 241-250 (1996)

Yoshihiro Shiba 等人：“非肽胃动素激动剂 EM523 对有意识的狗释放肠道和胰腺激素的影响。胃肠病学”。