A Novel Nitric Oxide Donor, S-Nitroso-NPivaloyl-D-Penicillamine, Activates a Non-Neuronal Cardiac Cholinergic System to Synthesize Acetylcholine and Augments Cardiac Function.
A Novel Nitric Oxide Donor, S-Nitroso-NPivaloyl-D-Penicillamine, Activates a Non-Neuronal Cardiac Cholinergic System to Synthesize Acetylcholine and Augments Cardiac Function.
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DOI：
10.33594/000000064
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发表时间：
2019-01-01
期刊：
影响因子：
--
通讯作者：
Kakinuma, Yoshihiko
中科院分区：
文献类型：
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作者：
Oikawa, Shino;Kai, Yuko;Kakinuma, Yoshihiko
背景/目的：在前期的研究中，我们报道了心肌细胞具有非神经性的心脏胆碱能系统（NNCCS）来合成乙酰胆碱（ACh），这是维持心脏基本生理功能所必需的。本研究的目的是确定和表征一个药理学诱导剂NNCCS.方法：为了确定一个药理学诱导剂NNCCS，我们筛选了几个化合物的化学结构类似的S-亚硝基-N-乙酰基-DL-青霉胺（SNAP）的结构。初步研究表明，SNAP是非神经元乙酰胆碱合成的诱导剂。我们筛选潜在的药理学诱导剂在H9 c2和HEK 293细胞中使用蛋白质印迹分析，荧光素酶测定，和测量细胞内cGMP，NO和ACh水平。结果：筛选出S-亚硝基-N-新戊酰-D-青霉胺（SNPiP），该化合物能逐渐升高H9 c2和HEK 293细胞内cGMP水平和一氧化氮（NO）水平。这些升高的水平导致胆碱乙酰转移酶基因的逐渐反式激活和翻译。此外，在体外和体内SNPiP治疗升高ACh水平72小时。SNPiP处理的小鼠上调了它们的心脏功能，没有心动过速，但舒张功能增强，导致心输出量改善。SNPiP的作用依赖于SNPiP的亚硝基基团，这通过缺乏亚硝基基团的N-特戊酰基-D-青霉胺（PiP）的无效性来验证。结论：SNPiP被鉴定为诱导NNCCS的重要药理学候选物之一。
BACKGROUND/AIMS: In a previous study, we reported that cardiomyocytes were equipped with non-neuronal cardiac cholinergic system (NNCCS) to synthesize acetylcholine (ACh), which is indispensable for maintaining the basic physiological cardiac functions. The aim of this study was to identify and characterize a pharmacological inducer of NNCCS.METHODS: To identify a pharmacological inducer of NNCCS, we screened several chemical compounds with chemical structures similar to the structure of S-nitroso-N-acetyl-DL-penicillamine (SNAP). Preliminary investigation revealed that SNAP is an inducer of non-neuronal ACh synthesis. We screened potential pharmacological inducers in H9c2 and HEK293 cells using western blot analysis, luciferase assay, and measurements of intracellular cGMP, NO₂ and ACh levels. The effects of the screened compound on cardiac function of male C57BL6 mice were also evaluated using cardiac catheter system.RESULTS: Among the tested compounds, we selected S-nitroso-Npivaloyl-D-penicillamine (SNPiP), which gradually elevated the intracellular cGMP levels and nitric oxide (NO) levels in H9c2 and HEK293 cells. These elevated levels resulted in the gradual transactivation and translation of the choline acetyltransferase gene. Additionally, in vitro and in vivo SNPiP treatment elevated ACh levels for 72 h. SNPiP-treated mice upregulated their cardiac function without tachycardia but with enhanced diastolic function resulting in improved cardiac output. The effect of SNPiP was dependent on SNPiP nitroso group as verified by the ineffectiveness of N-pivaloyl-D-penicillamine (PiP), which lacks the nitroso group.CONCLUSION: SNPiP is identified to be one of the important pharmacological candidates for induction of NNCCS.