Development of a Rapid Insulin Assay by Homogenous Time-Resolved Fluorescence.
Development of a Rapid Insulin Assay by Homogenous Time-Resolved Fluorescence.
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DOI：
10.1371/journal.pone.0148684
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Freyberg Z
中科院分区：
文献类型：
--
作者：
Farino ZJ;Morgenstern TJ;Vallaghe J;Gregor N;Donthamsetti P;Harris PE;Pierre N;Freyberg R;Charrier-Savournin F;Javitch JA;Freyberg Z
胰岛素的直接测量对于胰岛素分泌的基础和临床研究至关重要。然而，目前的方法是昂贵和耗时的。我们开发了一种基于均匀时间分辨荧光的胰岛素测定方法，该方法比目前常用的方法更快速，更具成本效益。该试验有效地应用于胰岛素分泌细胞系INS-1 E细胞以及胰岛，使我们能够通过阐明多巴胺调节胰岛素释放的机制来验证该试验。我们发现多巴胺在葡萄糖刺激的胰岛素分泌中起着重要的负调节作用。此外，我们发现，溴隐亭，一种已知的多巴胺D2/D3受体激动剂和新批准的用于治疗II型糖尿病的药物，也降低了葡萄糖刺激的胰岛胰岛素分泌水平与多巴胺治疗引起的水平相当。
Direct measurement of insulin is critical for basic and clinical studies of insulin secretion. However, current methods are expensive and time-consuming. We developed an insulin assay based on homogenous time-resolved fluorescence that is significantly more rapid and cost-effective than current commonly used approaches. This assay was applied effectively to an insulin secreting cell line, INS-1E cells, as well as pancreatic islets, allowing us to validate the assay by elucidating mechanisms by which dopamine regulates insulin release. We found that dopamine functioned as a significant negative modulator of glucose-stimulated insulin secretion. Further, we showed that bromocriptine, a known dopamine D2/D3 receptor agonist and newly approved drug used for treatment of type II diabetes mellitus, also decreased glucose-stimulated insulin secretion in islets to levels comparable to those caused by dopamine treatment.