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A Non-Targeted Capillary Electrophoresis-Mass Spectrometry Strategy to Study Metabolic Differences in an In vitro Model of High-Glucose Induced Changes in Human Proximal Tubular HK-2 Cells
Molecules ( IF 4.6 ) Pub Date : 2020-01-24 , DOI: 10.3390/molecules25030512
Samuel Bernardo-Bermejo 1 , Elena Sánchez-López 1, 2 , María Castro-Puyana 1, 2 , Selma Benito-Martínez 3, 4 , Francisco Javier Lucio-Cazaña 3 , María Luisa Marina 1, 2
Affiliation  

Diabetic nephropathy is characterized by the chronic loss of kidney function due to high glucose renal levels. HK-2 proximal tubular cells are good candidates to study this disease. The aim of this work was to study an in vitro model of high glucose-induced metabolic alterations in HK-2 cells to contribute to the pathogenesis of this diabetic complication. An untargeted metabolomics strategy based on CE-MS was developed to find metabolites affected under high glucose conditions. Intracellular and extracellular fluids from HK-2 cells treated with 25 mM glucose (high glucose group), with 5.5 mM glucose (normal glucose group), and with 5.5 mM glucose and 19.5 mM mannitol (osmotic control group) were analyzed. The main changes induced by high glucose were found in the extracellular medium where increased levels of four amino acids were detected. Three of them (alanine, proline, and glutamic acid) were exported from HK-2 cells to the extracellular medium. Other affected metabolites include Amadori products and cysteine, which are more likely cause and consequence, respectively, of the oxidative stress induced by high glucose in HK-2 cells. The developed CE-MS platform provides valuable insight into high glucose-induced metabolic alterations in proximal tubular cells and allows identifying discriminative molecules of diabetic nephropathy.

中文翻译:

一种非靶向毛细管电泳-质谱法研究高糖诱导人近端小管 HK-2 细胞变化体外模型中代谢差异的策略

糖尿病肾病的特征在于由于高葡萄糖肾水平导致肾功能的慢性丧失。HK-2 近端肾小管细胞是研究这种疾病的良好候选者。这项工作的目的是研究高糖诱导的 HK-2 细胞代谢改变的体外模型,以促进这种糖尿病并发症的发病机制。开发了一种基于 CE-MS 的非靶向代谢组学策略,以发现在高葡萄糖条件下受影响的代谢物。分析了来自用 25 mM 葡萄糖(高葡萄糖组)、5.5 mM 葡萄糖(正常葡萄糖组)和 5.5 mM 葡萄糖和 19.5 mM 甘露醇(渗透对照组)处理的 HK-2 细胞的细胞内和细胞外液。在检测到四种氨基酸水平增加的细胞外培养基中发现了高葡萄糖诱导的主要变化。其中三种(丙氨酸、脯氨酸和谷氨酸)从 HK-2 细胞输出到细胞外培养基。其他受影响的代谢物包括 Amadori 产物和半胱氨酸,它们更可能分别是 HK-2 细胞中高葡萄糖诱导的氧化应激的原因和结果。开发的 CE-MS 平台提供了对近端肾小管细胞中高葡萄糖诱导的代谢改变的宝贵见解,并允许识别糖尿病肾病的鉴别分子。
更新日期:2020-01-24
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