多巴胺受体在大鼠结肠黏膜下神经丛的表达和细胞分布

doi:10.3969/j.issn.1006-7795.2017.03.017

首都医科大学学报 ›› 2017, Vol. 38 ›› Issue (3): 411-416.doi: 10.3969/j.issn.1006-7795.2017.03.017

多巴胺受体在大鼠结肠黏膜下神经丛的表达和细胞分布

张悦^1,2, 李蕴¹, 朱进霞², 赵文明¹

1. 首都医科大学基础医学院免疫学系, 北京 100069;
2. 首都医科大学基础医学院生理学与病理生理学系, 北京 100069

收稿日期:2016-12-15 出版日期:2017-05-21 发布日期:2017-06-14
通讯作者: 李蕴, 赵文明 E-mail:liyun666@ccmu.edu.cn;zhao-wenming@163.com
基金资助:
国家自然科学基金（31300954，81370482）。

Expression and cellular distribution of dopamine receptors in the rat colonic submucosal plexus

Zhang Yue^1,2, Li Yun¹, Zhu Jinxia², Zhao Wenming¹

1. Department of Immunology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China;
2. Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China

Received:2016-12-15 Online:2017-05-21 Published:2017-06-14
Supported by:
This study was supported by National Natural Science Foundation of China (31300954, 81370482).

摘要/Abstract

摘要： 目的多巴胺（dopamine，DA）与其受体结合调节肠道动力、黏膜分泌及屏障等，黏膜下神经元参与DA对黏膜的生理调节，但其受体的具体分布尚不明了。通过检测DA受体在大鼠结肠黏膜下层的蛋白和mRNA表达及细胞分布，为DA调节大鼠结肠病理生理机制提供依据。方法用RT-PCR和Western blotting方法定性测定DA受体在大鼠结肠黏膜下层的mRNA和蛋白的表达；免疫荧光双染色后在激光共聚焦显微镜下观察DA受体在黏膜下血管活性肠肽（vasoactive intestinal peptide，VIP）能和胆碱能神经元上的分布。结果在大鼠结肠黏膜下层，DA受体D₁、D₂、D₅的mRNA及蛋白均有表达；在黏膜下神经丛VIP能阳性的神经元中有大量分布，其中D₁阳性的神经元占58.94%±2.245%，D₂阳性神经元占52.03%±9.384%，D₅阳性神经元占86.21%±2.902%，且D₅阳性神经元数量显著高于D₁和D₂；在黏膜下神经丛胆碱能阳性的神经元广泛表达DA受体，其中D₁阳性神经元占87.75%±7.307%，D₂阳性神经元占88.50%±8.761%，D₅阳性神经元占89.25%±10.75%。结论 DA受体在大鼠结肠黏膜下层有D₁、D₂及D₅的表达，且在VIP能及胆碱能神经元上均有分布，其中D₅受体在VIP能神经元上的分布显著高于D₁及D₂受体。本研究可为深入探讨DA调节结肠黏膜生理及病理机制提供形态学依据。

关键词: 多巴胺, 多巴胺受体, 黏膜下神经丛, 血管活性肠肽, 乙酰胆碱

Abstract: Objective Dopamine (DA) plays an important role in the regulation of intestinal motility, secretion and mucosal barrier by binding to its receptors. It has been shown that submucosal neurons are involved in the regulation effect of DA on the mucosa function. However the expressions of DA receptors in submucosal neurons are not clear. The aim of the present study is to investigate the distribution of DA receptors in vasoactive intestinal peptide (VIP) neurons and cholinergic neurons in the submucosa. Methods The mRNA and protein expression of DA receptors was measured by RT-PCR and Western blotting. Immunofluorescent double labeling of DA receptors with submucosal VIP neurons and cholinergic neurons were observed under confocal laser scanning microscope. Results Both the mRNA and protein of DA receptors D₁, D₂ and D₅ were expressed in the colonic submucosal layer. In the submucosal plexus, D₁ positive VIP-positive neurons accounting for 58.94%±2.245%, D₂ for 52.03%±9.384%, and D₅ for 86.21%±2.902%. The number of D₅ positive neurons was significantly higher than that of D₁ and D₂. In the submucosal plexus, D₁ positive cholinergic neurons accounted for 87.75%±7.307%, D₂ for 88.50%±8.761%, and D₅ for 89.25%±10.75%, and the distribution of the three types of DA receptors in the cholinergic neurons had no significant difference. Conclusion DA receptors D₁, D₂ and D₅ are expressed in the submucosal layer of the rat colon. And they all are distributed in the submucosal VIP and cholinergic neurons, and D₅ expression was higher than D₁ and D₂ in VIP-positive neurons. Our study provides a morphological basis for the regulation of DA on the colonic mucosal function.

Key words: dopamine, dopamine receptors, submucosal plexus, vasoactive intestinal peptide, acetylcholine

中图分类号:

R392

张悦, 李蕴, 朱进霞, 赵文明. 多巴胺受体在大鼠结肠黏膜下神经丛的表达和细胞分布[J]. 首都医科大学学报, 2017, 38(3): 411-416.

Zhang Yue, Li Yun, Zhu Jinxia, Zhao Wenming. Expression and cellular distribution of dopamine receptors in the rat colonic submucosal plexus[J]. Journal of Capital Medical University, 2017, 38(3): 411-416.

参考文献

[1] Pacheco R, Contreras F, Zouali M. The dopaminergic system in autoimmune diseases[J]. Front Immunol, 2014, 5:117.
[2] Tolstanova G, Deng X, Ahluwalia A, et al. Role of dopamine and D2 dopamine receptor in the pathogenesis of inflammatory bowel disease[J]. Dig Dis Sci, 2015,60(10):2963-2975.
[3] Zheng L F, Song J, Fan R F, et al. The role of vagal pathway and gastric dopamine in the gastroparesis of rats after 6-hydroxydopamine microinjection in the substantia nigra[J].Acta Physiol (Oxf),2014, 211(2):434-446.
[4] Feng X Y, Li Y, Li L S, et al. Dopamine D1 receptors mediate dopamine-induced duodenal epithelial ion transport in rats[J].Transl Res,2013,161(6):486-494.
[5] Zhang G H, Zhu J X, Xue H, et al. Dopamine stimulates Cl^- absorption coupled with HCO_{3 -} secretion in rat late distal colon[J]. Eur J Pharmacol, 2007,570(1-3):188-195.
[6] Al-Jahmany A A, Schultheiss G, Diener M. Effects of dopamine on ion transport across the rat distal colon[J].Pflugers Arch, 2004,448(6):605-612.
[7] Li Z S, Pham T D, Tamir H, et al. Enteric dopaminergic neurons:definition, developmental lineage, and effects of extrinsic denervation[J]. J Neurosci, 2004, 24(6):1330 -1339.
[8] Li Z S, Schmauss C, Cuenca A, et al. Physiological modulation of intestinal motility by enteric dopaminergic neurons and the D2 receptor:analysis of dopamine receptor expression, location, development, and function in wild-type and lnock-out mice[J]. J Neurosci, 2006,26(10):2798 -2807.
[9] Burns, A J, Hofstra R M. The enteric nervous system:From embryology to therapy[J]. Dev Biol, 2016,417(2):127-128.
[10] Foong J P, Tough I R, Cox H M, et al. Properties of cholinergic and non-cholinergic submucosal neurons along the mouse colon[J]. J Physiol, 2014,592(4):777-793.
[11] Zhang X, Li Y, Liu C, et al. Alteration of enteric monoamines with monoamine receptors and colonic dysmotility in 6-OHDA-induced Parkinson's disease rats[J].Transl Res,2015, 166(2):152-162.
[12] ZhangX H, Zhang X F, Zhang J Q, et al. β-adrenoceptors, but not dopamine receptors, mediate dopamine-induced ion transport in late distal colon of rats[J].Cell Tissue Res,2008,334(1):25-35.
[13] Chino Y, Fujimura M, Kitahama K, et al. Colocalization of NO and VIP in neurons of the submucous plexus in the rat intestine[J]. Peptides,2002, 23 (12):2245-2250.
[14] Gonzalez-Rey E, Delgado M. Therapeutic treatment of experimental colitis with regulatory dendritic cells generated with vasoactive intestinal peptide[J]. Gastroenterology, 2006, 131(6):1799-1811.
[15] Gonzalez-Rey E, Delgado M. Role of vasoactive intestinal peptide in inflammation and autoimmunity[J].Curr Opin Investig Drugs, 2005, 6(11):1116-1123.
[16] Reddix R, Kuhawara A, Wallace L, et al. Vasoactive intestinal polypeptide:a transmitter in submucous neurons mediating secretion in guinea pig distal colon[J]. J Pharmacol Exp Ther, 1994, 269(3):1124-1129.
[17] Ratcliffe E M, deSa D J, Dixon M F, et al. Intestines choline acetyltransferase (ChAT) immunoreactivity in paraffin sections of normal and diseased intestines[J].J Histochem Cytochem,1998, 46(11):1223-1231.
[18] Kawano M, Takagi R, Kaneko A, et al. Berberine is a dopamine D1-and D2-like receptor antagonist and ameliorates experimentally induced colitis by suppressing innate and adaptive immune responses[J]. J Neuroimmunol, 2015, 289:43-55.
[19] Tolstanova G, Deng X, Ahluwalia A, et al. Role of dopamine and D2 dopamine receptor in the pathogenesis of inflammatory bowel disease[J]. Dig Dis Sci, 2015, 60(10):2963-2975.
[20] Magro F, Vieira-Coelho M A, Fraga S, et al. Impaired synthesis or cellular storage of norepinephrine, dopamine, and 5-hydroxytryptamine in human inflammatory bowel disease[J]. Dig Dis Sci, 2002, 47(1):216-224.

多巴胺受体在大鼠结肠黏膜下神经丛的表达和细胞分布

Expression and cellular distribution of dopamine receptors in the rat colonic submucosal plexus

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