Metabolome response to glucose in the β-cell line INS-1 832/13.
Metabolome response to glucose in the β-cell line INS-1 832/13.
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DOI：
10.1074/jbc.m112.414961
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发表时间：
2013-04-12
期刊：
影响因子：
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通讯作者：
Burant CF
中科院分区：
文献类型：
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作者：
Lorenz MA;El Azzouny MA;Kennedy RT;Burant CF
背景：葡萄糖刺激胰岛素分泌的生化途径尚未完全阐明。结果：质谱分析显示，葡萄糖在INS-1细胞中引起了快速而实质性的代谢重编程。结论：代谢组学允许测试和产生关于葡萄糖在胰岛素分泌细胞中的作用的多种假设。意义：了解葡萄糖刺激胰岛素分泌的生化基础对了解2型糖尿病的根本原因至关重要。胰β细胞葡萄糖刺激胰岛素分泌（GSIS）是由糖代谢引起的，增加ATP/ADP比值，阻断KATP通道，导致膜去极化和胰岛素胞吐。其他被认为增加胰岛素分泌的代谢途径尚未完全阐明。为了研究GSIS期间的代谢变化，采用液相色谱-质谱法测定了在葡萄糖从3到10 mm葡萄糖变化以及INS-1 832/13 β-细胞系中葡萄糖浓度增加时的87种代谢物水平。U-[13C]葡萄糖用于探测特定代谢途径中的通量。结果包括ATP/ADP快速增加，三羧酸循环通量和丙二醇辅酶a途径增加，支持GSIS的流行理论。新的发现包括：用于补钙的天冬氨酸并非来源于葡萄糖燃料，葡萄糖被添加到刺激胰岛素分泌、葡萄糖通量进入甘油-3-磷酸、长链辅酶a的酯化反应导致长链辅酶a的快速消耗和磷脂酸和二酰基甘油的重新生成。此外，在GSIS中具有潜在作用的新代谢物，如5-氨基咪唑-4-羧酰胺核糖肽（ZMP）， gdp -甘露糖和焦磷酸法尼酯被发现在葡萄糖暴露后迅速改变。
Background: The biochemical pathways underlying glucose-stimulated insulin secretion have not been fully elucidated. Results: Mass spectrometry analysis revealed rapid and substantial metabolic reprogramming evoked by glucose in INS-1 cells. Conclusion: Metabolomics allowed testing and generation of multiple hypotheses regarding glucose effects in insulin-secreting cells. Significance: Insights into the biochemical basis of glucose-stimulated insulin secretion are critical for understanding root causes of type 2 diabetes. Glucose-stimulated insulin secretion (GSIS) from pancreatic β-cells is triggered by metabolism of the sugar to increase ATP/ADP ratio that blocks the KATP channel leading to membrane depolarization and insulin exocytosis. Other metabolic pathways believed to augment insulin secretion have yet to be fully elucidated. To study metabolic changes during GSIS, liquid chromatography with mass spectrometry was used to determine levels of 87 metabolites temporally following a change in glucose from 3 to 10 mm glucose and in response to increasing concentrations of glucose in the INS-1 832/13 β-cell line. U-[13C]Glucose was used to probe flux in specific metabolic pathways. Results include a rapid increase in ATP/ADP, anaplerotic tricarboxylic acid cycle flux, and increases in the malonyl CoA pathway, support prevailing theories of GSIS. Novel findings include that aspartate used for anaplerosis does not derive from the glucose fuel added to stimulate insulin secretion, glucose flux into glycerol-3-phosphate, and esterification of long chain CoAs resulting in rapid consumption of long chain CoAs and de novo generation of phosphatidic acid and diacylglycerol. Further, novel metabolites with potential roles in GSIS such as 5-aminoimidazole-4-carboxamide ribotide (ZMP), GDP-mannose, and farnesyl pyrophosphate were found to be rapidly altered following glucose exposure.