H. Cabadak, E. Küçükibrahimoğlu, B. Aydın, B. Kan, M. Zafer Gören
{"title":"Muscarinic receptor-mediated nitric oxide release in a K562 erythroleukaemia cell line","authors":"H. Cabadak, E. Küçükibrahimoğlu, B. Aydın, B. Kan, M. Zafer Gören","doi":"10.1111/j.1474-8673.2009.00431.x","DOIUrl":null,"url":null,"abstract":"<div>\n \n <p> <b>1</b> In the present study we have investigated the expression of muscarinic receptors in K562 erythroleukaemic cells and the effects of muscarinic agonist and antagonists on extracellular citrulline levels in these cells, as a marker of nitric oxide (NO) generation.</p>\n <p> <b>2</b> Muscarinic acetylcholine receptors (M<sub>1</sub>–M<sub>5</sub>) play key roles in regulating many diverse physiological processes. Recent studies suggest that muscarinic receptors mediate some cellular events in haematopoietic cells. Multiple subtypes of muscarinic receptors are expressed in different human cells. NO, a free radical and a signaling molecule, is involved in the regulation of many physiological functions and derived from certain nitric oxide synthases (NOS), which are related to muscarinic receptors.</p>\n <p> <b>3</b> In this study, the presence of M<sub>2</sub>, M<sub>3</sub> and M<sub>4</sub> subtypes in K562, an erythroleukaemic cell line, was demonstrated by using the reverse transcriptase-polymerase chain reaction. Moreover, the generation of NO induced by carbachol, a non-selective muscarinic agonist, was investigated by using high-performance liquid chromatography to measure changes in extracellular <span>l</span>-citrulline levels.</p>\n <p> <b>4</b> We found that carbachol enhanced <span>l</span>-citrulline production in K562 erythroleukaemic cells. The effect of carbachol on <span>l</span>-citrulline production was antagonized by atropine and 4-diphenylacetoxy-N-methylpiperidine (4-DAMP), while tropicamide had little effect. These results suggest that the muscarinic receptor M<sub>3</sub> subtype may mediate NO signaling in K562 erythroleukaemic cells.</p>\n </div>","PeriodicalId":100151,"journal":{"name":"Autonomic and Autacoid Pharmacology","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2009-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1111/j.1474-8673.2009.00431.x","citationCount":"12","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Autonomic and Autacoid Pharmacology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1111/j.1474-8673.2009.00431.x","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 12
Abstract
1 In the present study we have investigated the expression of muscarinic receptors in K562 erythroleukaemic cells and the effects of muscarinic agonist and antagonists on extracellular citrulline levels in these cells, as a marker of nitric oxide (NO) generation.
2 Muscarinic acetylcholine receptors (M1–M5) play key roles in regulating many diverse physiological processes. Recent studies suggest that muscarinic receptors mediate some cellular events in haematopoietic cells. Multiple subtypes of muscarinic receptors are expressed in different human cells. NO, a free radical and a signaling molecule, is involved in the regulation of many physiological functions and derived from certain nitric oxide synthases (NOS), which are related to muscarinic receptors.
3 In this study, the presence of M2, M3 and M4 subtypes in K562, an erythroleukaemic cell line, was demonstrated by using the reverse transcriptase-polymerase chain reaction. Moreover, the generation of NO induced by carbachol, a non-selective muscarinic agonist, was investigated by using high-performance liquid chromatography to measure changes in extracellular l-citrulline levels.
4 We found that carbachol enhanced l-citrulline production in K562 erythroleukaemic cells. The effect of carbachol on l-citrulline production was antagonized by atropine and 4-diphenylacetoxy-N-methylpiperidine (4-DAMP), while tropicamide had little effect. These results suggest that the muscarinic receptor M3 subtype may mediate NO signaling in K562 erythroleukaemic cells.
在本研究中,我们研究了毒蕈碱受体在K562红白血病细胞中的表达,以及毒蕈碱激动剂和拮抗剂对这些细胞外瓜氨酸水平的影响,瓜氨酸是一氧化氮(NO)生成的标志。毒蕈碱乙酰胆碱受体(M1-M5)在调节多种生理过程中起关键作用。最近的研究表明,毒蕈碱受体介导造血细胞的一些细胞事件。毒蕈碱受体的多种亚型在不同的人类细胞中表达。一氧化氮(NO)是一种自由基和信号分子,参与许多生理功能的调节,来源于某些与毒蕈碱受体有关的一氧化氮合酶(NOS)。3本研究通过逆转录-聚合酶链反应证实了红白血病细胞系K562中存在M2、M3和M4亚型。此外,采用高效液相色谱法测定细胞外l-瓜氨酸水平的变化,研究了非选择性毒菌碱激动剂carbachol诱导NO的生成。4 .我们发现,乙醇能增强K562红白血病细胞中l-瓜氨酸的生成。阿托品和4-二苯基乙酰氧基- n -甲基哌啶(4-DAMP)可拮抗卡巴醇对l-瓜氨酸生成的影响,而tropicamide对其影响不大。这些结果提示毒蕈碱受体M3亚型可能介导K562红白血病细胞NO信号转导。