Evaluation of tauroursodeoxycholic acid in liver cells’ cultures by MEKC: Initial hints to comprehend its role in diabetes mellitus of obese individuals

IF 3 3区生物学 Q2 BIOCHEMICAL RESEARCH METHODS ELECTROPHORESIS Pub Date : 2024-05-22 DOI:10.1002/elps.202300223

Bruna Eduarda Santos Simões, Mariana Roberta Rodrigues Muniz, Thiago dosReis Araujo, Everardo Magalhães Carneiro, Ana Valéria Colnaghi Simionato

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Abstract

Genetic factors, diet, lifestyle, and other factors lead to various complications in the body, such as obesity and other chronic diseases. The inflammatory state caused by excessive accumulation of body fat affects the pathways related to the control of glycemic homeostasis, leading to a high demand for insulin, to subsequent failure of stressed β cells, and development of type 2 diabetes mellitus (T2DM). The study of new endocrine signalers, such as bile acids (BAs), becomes necessary as it allows the development of alternatives for T2DM treatment. In this work, a methodology was developed to quantify tauroursodeoxycholic BA (TUDCA) in liver cells of the HepG2 strain treated in hyperlipidic medium. This BA helps to improve insulin clearance by increasing the expression of the insulin-degrading enzyme, restoring sensitivity to this hormone, and making it viable for treating T2DM. Herein, a targeted metabolomic method for TUDCA determination in extracellular medium of hepatocyte matrices by micellar electrokinetic chromatography-UV was optimized, validated, and applied. The optimized background electrolyte was composed of 40 mmol/L sodium cholate and 30 mmol/L sodium tetraborate at pH 9.0. The following figures of merit were evaluated: linearity, limit of quantification, limit of detection, accuracy, and precision. Data obtained with the validated electrophoretic method showed a self-stimulation of TUDCA production in media supplemented only with BA. On the other hand, TUDCA concentration was reduced in the hyperlipidic medium. This suggests that, in these media, the effect of TUDCA is reduced, such as self-stimulated production and consequent regulation of glycemic homeostasis. Therefore, the results reinforce the need for investigating TUDCA as a potential T2DM biomarker as well as its use to treat several comorbidities, such as obesity and diabetes mellitus.

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用 MEKC 评估肝细胞培养物中的牛磺脱氧胆酸：初步了解其在肥胖者糖尿病中的作用。

遗传因素、饮食、生活方式和其他因素导致人体出现各种并发症，如肥胖和其他慢性疾病。体内脂肪过度积聚引起的炎症状态会影响与控制血糖平衡相关的途径，导致对胰岛素的高需求，进而使受压的 β 细胞失效，发展成 2 型糖尿病（T2DM）。对胆汁酸（BA）等新的内分泌信号物质的研究变得十分必要，因为这有助于开发治疗 T2DM 的替代品。在这项工作中，我们开发了一种方法来量化在高脂培养基中处理的 HepG2 株肝细胞中的牛磺脱氧胆汁酸（TUDCA）。这种 BA 有助于通过增加胰岛素降解酶的表达来改善胰岛素清除率，恢复对这种激素的敏感性，使其成为治疗 T2DM 的可行方法。本文对胶束电动色谱-紫外法测定肝细胞基质细胞外培养基中TUDCA的靶向代谢组学方法进行了优化、验证和应用。优化后的背景电解质由 40 mmol/L 胆酸钠和 30 mmol/L 四硼酸钠组成，pH 值为 9.0。对以下指标进行了评估：线性度、定量限、检测限、准确度和精密度。使用经验证的电泳方法获得的数据表明，在仅添加 BA 的培养基中，TUDCA 的产生具有自我刺激作用。另一方面，高脂培养基中的 TUDCA 浓度降低。这表明，在这些培养基中，TUDCA 的作用会减弱，如自我刺激产生，进而调节血糖平衡。因此，这些结果加强了研究 TUDCA 作为潜在 T2DM 生物标志物的必要性，以及将其用于治疗肥胖症和糖尿病等几种合并症的必要性。

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来源期刊

ELECTROPHORESIS 生物-分析化学

CiteScore

6.30

自引率

13.80%

发文量

244

审稿时长

1.9 months

期刊介绍： ELECTROPHORESIS is an international journal that publishes original manuscripts on all aspects of electrophoresis, and liquid phase separations (e.g., HPLC, micro- and nano-LC, UHPLC, micro- and nano-fluidics, liquid-phase micro-extractions, etc.). Topics include new or improved analytical and preparative methods, sample preparation, development of theory, and innovative applications of electrophoretic and liquid phase separations methods in the study of nucleic acids, proteins, carbohydrates natural products, pharmaceuticals, food analysis, environmental species and other compounds of importance to the life sciences. Papers in the areas of microfluidics and proteomics, which are not limited to electrophoresis-based methods, will also be accepted for publication. Contributions focused on hyphenated and omics techniques are also of interest. Proteomics is within the scope, if related to its fundamentals and new technical approaches. Proteomics applications are only considered in particular cases. Papers describing the application of standard electrophoretic methods will not be considered. Papers on nanoanalysis intended for publication in ELECTROPHORESIS should focus on one or more of the following topics: • Nanoscale electrokinetics and phenomena related to electric double layer and/or confinement in nano-sized geometry • Single cell and subcellular analysis • Nanosensors and ultrasensitive detection aspects (e.g., involving quantum dots, "nanoelectrodes" or nanospray MS) • Nanoscale/nanopore DNA sequencing (next generation sequencing) • Micro- and nanoscale sample preparation • Nanoparticles and cells analyses by dielectrophoresis • Separation-based analysis using nanoparticles, nanotubes and nanowires.