Departamento de Química Analítica, Química Física e Ingeniería Química, Universidad de Alcalá, Ctra. Madrid-Barcelona Km. 33.600, 28871 Madrid, Spain.
Department of Human Genetics, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands.
Int J Mol Sci. 2021 Jul 9;22(14):7399. doi: 10.3390/ijms22147399.
Oxygen deficiency in cells, tissues, and organs can not only prevent the proper development of biological functions but it can also lead to several diseases and disorders. In this sense, the kidney deserves special attention since hypoxia can be considered an important factor in the pathophysiology of both acute kidney injury and chronic kidney disease. To provide better knowledge to unveil the molecular mechanisms involved, new studies are necessary. In this sense, this work aims to study, for the first time, an in vitro model of hypoxia-induced metabolic alterations in human proximal tubular HK-2 cells because renal proximal tubules are particularly susceptible to hypoxia. Different groups of cells, cultivated under control and hypoxia conditions at 0.5, 5, 24, and 48 h, were investigated using untargeted metabolomic approaches based on reversed-phase liquid chromatography-mass spectrometry. Both intracellular and extracellular fluids were studied to obtain a large metabolite coverage. On the other hand, multivariate and univariate analyses were carried out to find the differences among the cell groups and to select the most relevant variables. The molecular features identified as affected metabolites were mainly amino acids and Amadori compounds. Insights about their biological relevance are also provided.
细胞、组织和器官缺氧不仅会妨碍生物功能的正常发育，还会导致多种疾病和紊乱。在这方面，肾脏尤其值得关注，因为缺氧可被视为急性肾损伤和慢性肾病病理生理学的一个重要因素。为了提供更好的知识来揭示所涉及的分子机制，有必要开展新的研究。在这方面，本工作首次旨在研究人近端肾小管 HK-2 细胞缺氧诱导的代谢改变的体外模型，因为肾脏近端小管特别容易受到缺氧的影响。采用基于反相液相色谱-质谱的非靶向代谢组学方法，研究了在对照和 0.5、5、24 和 48 h 缺氧条件下培养的不同细胞组。研究了细胞内和细胞外液，以获得广泛的代谢物覆盖。另一方面，进行了多变量和单变量分析，以找到细胞组之间的差异，并选择最相关的变量。确定为受影响代谢物的分子特征主要是氨基酸和麦拉德化合物。还提供了关于其生物学相关性的见解。
