遗传因素,饮食,生活方式,和其他因素导致体内各种并发症,如肥胖和其他慢性疾病。体内脂肪过度积累引起的炎症状态影响与控制血糖稳态相关的途径,导致对胰岛素的高需求,随后应激β细胞的失败,2型糖尿病(T2DM)的发生发展。新的内分泌信号的研究,如胆汁酸(BA),由于它允许开发T2DM治疗的替代方案,因此变得必要。在这项工作中,开发了一种方法来定量在高脂培养基中处理的HepG2菌株的肝细胞中的牛磺熊去氧胆酸BA(TUDCA)。这种BA有助于通过增加胰岛素降解酶的表达来提高胰岛素清除率,恢复对这种激素的敏感性,并使其适用于治疗T2DM。在这里,优化了通过胶束电动色谱-UV测定肝细胞基质细胞外培养基中TUDCA的靶向代谢组学方法,已验证,并应用。优化的背景电解质由pH为9.0的40mmol/L胆酸钠和30mmol/L四硼酸钠组成。评估了以下优值:线性,定量极限,检测限,准确度,和精度。通过验证的电泳方法获得的数据显示,在仅补充有BA的培养基中,TUDCA生产的自我刺激。另一方面,TUDCA浓度在高脂培养基中降低。这表明,在这些媒体中,TUDCA的效果降低,如自我刺激的生产和随之而来的调节血糖稳态。因此,结果加强了对TUDCA作为潜在的T2DM生物标志物以及其用于治疗多种合并症的研究的需求,如肥胖和糖尿病。
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.