Motilin release in perifused isolated canine duodenal epithelial cells.

灌注分离的犬十二指肠上皮细胞中胃动素的释放。

基本信息

批准号：
06670520
负责人：
SAKAI Takafumi
金额：
$ 1.34万
依托单位：
Saitama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
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-06670520/
关键词：
Motilin Perifusion System Acetylcholine Muscarinic Receptors Dog Duodenal epithelial cells

项目摘要

In spite of numerous invesigations devoted to the motor stimulatory effect of motilin on gastrointestinal smooth muscle, little is known regarding the mechanisms controlling the release of motilin from its producing cells. The present study was designed to 1) develop a perifusion system with dispersed canine duodenal epithelial cells, and 2) assess the effect of acetylcholine (ACh), somatostatin and bombesin on motilin release, and 3) determine the muscarinic receptor subtypes on the motilin producing cells.Segments of the duodenum were dissected out from anesthetized dogs, and the mucosal layr was removed from the muscle layr with fine scissors. Isolated epithelial cells were obtained by collagenase dispersion and percoll separation. The dispersed cells were mixed with preswollen Bio-Gel P2 and aliquoted into columns (1 ml volume). The perifusion medium (HEPES buffered Hank's solution) was kept at 37ﾟC and continuously bubbled with a 100% O2. The flow rate was 0.3 ml/min, the fraction … More s were collected at 3 min intervals, and the motilin concentration was determined by double-antibody radio-immunoassay. Carbachol (Cch), Ach agonist, (10^<-10>M to 10^<-1>M) stimulated release of motilin from the isolated cell column in a dose-dependent manner with the maximum response at 10^<-4>M.This response to Cch was abolished by atropine (10^<-4>M), but somatostatin (10^<-4>M) had no effect on the ACh-stimulated motilin release. Bombesin also had no effect on motilin release in the concentration range tested (10^<-10>M to 10^<-6>M). Pirenzepine (M1 receptor agonist) and AF-DX116BS (M2 receptor agonist) had no effect on Cch (10^<-4>M) Stimulated motilin release at 10^<-6>M,but 4-DAMP (M3 receptor agonist) (10^<-6>M) completely abolished Cch-stimulated motilin release.Conclusion : 1) A perifusion system which can monitor rapid motilin release was established. 2) Ach directly stimulates motilin release though M3 receptors on the cell.These results indicate that release of motilin from the cells are regulated by the colinergic neural stimulation. Less

尽管进行了许多研究，这些研究致力于Motilin对胃肠道平滑肌的运动刺激作用，但对于控制Motilin从其生产细胞中释放的机制而言，知之甚少。 The present study was designed to 1) developing a perifusion system with dispersed canine duodenal epithelial cells, and 2) assessing the effect of acetylcholine (ACh), somatostatin and bombesin on motilin release, and 3) determining the muscarinic receptor subtypes on the motilin producing cells.Segments of the duodenum were dissected out from anesthetized dogs, and the mucosal layr was用精细的剪刀从肌肉左上移走。通过胶原酶分散和珀托分离获得分离的上皮细胞。将分散的细胞与前沃伦生物凝胶P2混合，并将其等分为色谱柱（1 ml体积）。将炎症培养基（HEPES缓冲汉克溶液）保持在37°C，并用100％O2连续冒泡。流速为0.3 mL/min，以3分钟的间隔收集了更多的s，并通过双重抗体射线免疫测定确定motilin浓度。 Carbachol (Cch), Ach agonist, (10^<-10>M to 10^<-1>M) stimulated release of motilin from the isolated cell column in a dose-dependent manner with the maximum response at 10^<-4>M.This response to Cch was abolished by atropine (10^<-4>M), but somatostatin (10^<-4>M) had no effect on the ACh-stimulated motilin release.在测试的浓度范围内（10^<-10> m至10^<-6> m），孟买对Motilin释放也没有影响。 Pirenzepine (M1 receptor agonist) and AF-DX116BS (M2 receptor agonist) had no effect on Cch (10^<-4>M) Stimulated motilin release at 10^<-6>M, but 4-DAMP (M3 receptor agonist) (10^<-6>M) completely abolished Cch-stimulated motilin release.Conclusion : 1) A perifusion system which can monitor rapid motilin release was 已确立的。 2）ACH直接刺激细胞上的M3受体释放Motilin蛋白。这些结果表明，从细胞中释放Motilin释放受到结链神经刺激的调节。较少的