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

尽管针对胃动素对胃肠道平滑肌的运动刺激作用进行了大量研究，但对于控制胃动素从其产生细胞中释放的机制知之甚少。本研究旨在1)开发具有分散的犬十二指肠的灌注系统。上皮细胞，2) 评估乙酰胆碱 (ACh)、生长抑素和铃蟾肽对胃动素释放的影响，3) 确定胃动素产生细胞上的毒蕈碱受体亚型。从麻醉狗身上切下十二指肠节段，用细剪刀将粘膜层从肌肉层上除去，通过胶原酶分散和percoll分离获得分离的上皮细胞。与预溶胀的 Bio-Gel P2 混合并等分到柱中（1 ml 体积）。将培养基（HEPES 缓冲的 Hank's 溶液）保持在 37°C，并用 100% O2 连续鼓泡，流速为 0.3 ml/min，以 3 分钟的间隔收集馏分，并通过下式测定胃动素浓度。双抗体放射免疫测定法。Carbachol (Cch)，Ach 激动剂，(10^<-10>M 至 10^<-1>M)以剂量依赖性方式刺激胃动素从分离的细胞柱中释放，最大反应在10^-4M。这种对Cch的反应被阿托品(10^-4M)消除，但生长抑素(10 ^ -4 M)对ACh刺激的胃动素释放没有影响，铃蟾肽在测试的浓度范围(10^ -10 M至10^ -10 M)内对胃动素释放也没有影响。 10^-6M)哌仑西平(M1受体激动剂)和AF-DX116BS(M2受体激动剂)对Cch(10^-4M)刺激的胃动素释放没有影响，在10^-6M下。但4-DAMP（M3受体激动剂）（10^-6M）完全消除了Cch刺激的胃动素释放。结论：1）A建立了可监测快速胃动素释放的灌注系统。2)Ach通过细胞上的M3受体直接刺激胃动素释放。这些结果表明细胞中胃动素的释放受共碱能神经刺激的调节。