Regulation of neurotensin release from canine enteric primary cell cultures.
Regulation of neurotensin release from canine enteric primary cell cultures.
复制标题
犬肠道原代细胞培养物神经降压素释放的调节。
DOI：
10.1152/ajpgi.1986.250.3.g385
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Soll,AH
中科院分区：
文献类型：
--
作者：
Barber,DL;Buchan,AM;Walsh,JH;Soll,AH
最近开发的原代细胞培养系统可以直接研究调节肠粘膜细胞神经降压素分泌的细胞机制。我们现在报告使用这些方法来评估肾上腺素能、胆碱能和肽能递质对神经降压素释放的调节作用。胶原酶分散的犬回肠黏膜细胞经离心洗脱后富含神经降压素样免疫反应(NTLI)，在胶原蛋白包裹的培养皿中培养48h。肾上腺素(0.01-100微米)刺激NTLI分泌呈剂量依赖性增加。NTLI对肾上腺素的反应增加了NTLI的分泌。心得安竞争性抑制NTLI对肾上腺素的反应，使肾上腺素剂量-反应曲线平行右移。α-肾上腺素能激动剂甲氧胺(10微米)和可乐定(10微米)不改变NTLI的基础分泌。肾上腺素能拮抗剂哌唑嗪(10微米)或育亨宾(10微米)对肾上腺素刺激无明显抑制作用。腺苷环化酶激活剂Forskolin二萜可增加NTLI的释放，并对肾上腺素的反应有相加作用。与β-肾上腺素能激活相反，卡巴胆碱和生长抑素对肾上腺素刺激的NTLI释放有剂量依赖性的抑制作用。在100微米卡巴胆碱时，NTLI的释放被抑制68%，这一作用可被阿托品(0.1微米)部分阻断。生长抑素(100 NM)产生96%的抑制，这是1 mM肾上腺素所不能克服的。这些数据表明，神经降压素的释放受到β-肾上腺素能激动剂和腺苷环化酶激活的刺激。生长抑素和M受体激动剂卡巴胆碱直接抑制NTLI的释放。
A recently developed primary cell-culture system allows direct study of the cellular mechanisms regulating neurotensin secretion from intestinal mucosal cells. We now report the use of these methods to evaluate the modulation of neurotensin release by adrenergic, cholinergic, and peptidergic transmitters. Collagenase-dispersed canine ileal mucosal cells, enriched for neurotensinlike immunoreactivity (NTLI) by centrifugal elutriation, were maintained for 48 h on collagen-coated culture dishes. Epinephrine (0.01-100 microM) stimulated a dose-dependent increase increase in NTLI secretion. The NTLI response to epinephrine increase in NTLI secretion. The NTLI response to epinephrine was competitively inhibited by propranolol, producing a parallel rightward shift of the epinephrine dose-response curve. alpha-Adrenergic agonist methoxamine (10 microM) and clonidine (10 microM) did not alter basal NTLI secretion. Epinephrine stimulation was not significantly inhibited by the alpha-adrenergic antagonists prazosin (10 microM) or yohimbine (10 microM). The diterpene forskolin, an adenyl cyclase activator, increased NTLI release and had an additive effect on the response to epinephrine. In contrast to beta-adrenergic activation, carbachol and somatostatin produced a dose-dependent inhibition of epinephrine-stimulated NTLI release. At 100 microM carbachol, NTLI release was inhibited 68%, and this action was partially blocked by atropine (0.1 microM). Somatostatin (100 nM) produced a 96% inhibition that was not surmountable by 1 mM epinephrine. These data indicate that neurotensin release is stimulated by beta-adrenergic agonists and adenylate cyclase activation. Somatostatin and the muscarinic agonist carbachol directly inhibit NTLI release.