Division of Pathological Sciences, Department of Pharmacology and Experimental Therapeutics, Kyoto Pharmaceutical University, Misasagi, Yamashina, Kyoto, Japan.
Department of Gastroenterology and Metabology, Ehime University Graduate School of Medicine, Toon, Ehime, Japan.
PLoS One. 2019 Feb 21;14(2):e0205939. doi: 10.1371/journal.pone.0205939. eCollection 2019.
Motilin is a gastrointestinal peptide hormone that stimulates gastrointestinal motility. Motilin is produced primarily in the duodenum and jejunum. Motilin receptors (MTLRs) are G protein-coupled receptors that may represent a clinically useful pharmacological target as they can be activated by erythromycin. The functions of motilin are highly species-dependent and remain poorly understood. As a functional motilin system is absent in rodents such as rats and mice, these species are not commonly used for basic studies. In this study, we examine the usefulness of human MTLR-overexpressing transgenic (hMTLR-Tg) mice by identifying the mechanisms of the gastric motor response to human motilin and erythromycin. The distribution of hMTLR was examined immunohistochemically in male wild-type (WT) and hMTLR-Tg mice. The contractile response of gastric strips was measured isometrically in an organ bath, while gastric emptying was determined using phenol red. hMTLR expression was abundant in the gastric smooth muscle layer. Interestingly, higher levels of hMTLR expression were observed in the myenteric plexus of hMTLR-Tg mice but not WT mice. hMTLR was not co-localized with vesicular acetylcholine transporter, a marker of cholinergic neurons in the myenteric plexus. Treatment with human motilin and erythromycin caused concentration-dependent contraction of gastric strips obtained from hMTLR-Tg mice but not from WT mice. The contractile response to human motilin and erythromycin in hMTLR-Tg mice was affected by neither atropine nor tetrodotoxin and was totally absent in Ca2+-free conditions. Furthermore, intraperitoneal injection of erythromycin significantly promoted gastric emptying in hMTLR-Tg mice but not in WT mice. Human motilin and erythromycin stimulate gastric smooth muscle contraction in hMTLR-Tg mice. This action is mediated by direct contraction of smooth muscle via the influx of extracellular Ca2+. Thus, hMTLR-Tg mice may be useful for the evaluation of MTLR agonists as gastric prokinetic agents.
胃动素是一种胃肠道肽激素,可刺激胃肠道蠕动。胃动素主要在十二指肠和空肠中产生。胃动素受体(MTLRs)是 G 蛋白偶联受体,它们可能代表一个有临床应用价值的药理学靶点,因为它们可以被红霉素激活。胃动素的功能高度依赖于物种,目前仍知之甚少。由于像大鼠和小鼠这样的啮齿动物中没有功能性胃动素系统,因此这些物种通常不用于基础研究。在这项研究中,我们通过鉴定人胃动素和红霉素引起胃运动反应的机制,来研究人 MTLR 过表达转基因(hMTLR-Tg)小鼠的有用性。我们通过免疫组织化学方法检测雄性野生型(WT)和 hMTLR-Tg 小鼠中 hMTLR 的分布。我们在器官浴中通过等长测量胃条的收缩反应,同时使用酚红测定胃排空。hMTLR 在胃平滑肌层中表达丰富。有趣的是,在 hMTLR-Tg 小鼠的肌间神经丛中观察到更高水平的 hMTLR 表达,但在 WT 小鼠中没有观察到。hMTLR 与囊泡乙酰胆碱转运体(肌间神经丛中胆碱能神经元的标志物)没有共定位。用人类胃动素和红霉素处理可引起来自 hMTLR-Tg 小鼠而不是 WT 小鼠的胃条产生浓度依赖性收缩。hMTLR-Tg 小鼠对人胃动素和红霉素的收缩反应不受阿托品或河豚毒素的影响,并且在无钙条件下完全消失。此外,红霉素腹腔注射可显著促进 hMTLR-Tg 小鼠的胃排空,但不能促进 WT 小鼠的胃排空。人类胃动素和红霉素可刺激 hMTLR-Tg 小鼠的胃平滑肌收缩。这种作用是通过细胞外 Ca2+ 内流直接收缩平滑肌介导的。因此,hMTLR-Tg 小鼠可能有助于评估 MTLR 激动剂作为胃动力药物。