Department of Chemistry, Indian Institute of Science Education and Research (IISER) Pune, Dr. Homi Bhabha Road, Pune, Maharashtra, 411008, India.
Department of Biotechnology, Savitribai Phule Pune University (Formerly University of Pune), Pune, Maharashtra, 411007, India.
Metabolomics. 2019 Mar 29;15(4):55. doi: 10.1007/s11306-019-1516-3.
Chronic exposure to high-glucose and free fatty acids (FFA) alone/or in combination; and the resulting gluco-, lipo- and glucolipo-toxic conditions, respectively, have been known to induce dysfunction and apoptosis of β-cells in Diabetes. The molecular mechanisms and the development of biomarkers that can be used to predict similarities and differences behind these conditions would help in easier and earlier diagnosis of Diabetes.
This study aims to use metabolomics to gain insight into the mechanisms by which β-cells respond to excess-nutrient stress and identify associated biomarkers.
INS-1E cells were cultured in high-glucose, palmitate alone/or in combination for 24 h to mimic gluco-, lipo- and glucolipo-toxic conditions, respectively. Biochemical and cellular experiments were performed to confirm the establishment of these conditions. To gain molecular insights, abundant metabolites were identified and quantified using H-NMR.
No loss of cellular viability was observed in high-glucose while exposure to FFA alone/in combination with high-glucose was associated with increased ROS levels, membrane damage, lipid accumulation, and DNA double-strand breaks. Forty-nine abundant metabolites were identified and quantified using H-NMR. Chemometric pair-wise analysis in glucotoxic and lipotoxic conditions, when compared with glucolipotoxic conditions, revealed partial overlap in the dysregulated metabolites; however, the dysregulation was more significant under glucolipotoxic conditions.
The current study compared gluco-, lipo- and glucolipotoxic conditions in parallel and elucidated differences in metabolic pathways that play major roles in Diabetes. o-phosphocholine and UDP-N-acetylglucosamine were identified as common dysregulated metabolites and their ratio was proposed as a potential biomarker for these conditions.
长期慢性暴露于高浓度葡萄糖和游离脂肪酸（FFA）中，或者同时暴露于高浓度葡萄糖和 FFA 中，分别会导致β细胞的糖毒性、脂毒性和糖脂毒性。这些条件下β细胞功能障碍和凋亡的分子机制以及可用于预测这些条件相似性和差异性的生物标志物的发展，将有助于糖尿病的早期诊断。
本研究旨在使用代谢组学来深入了解β细胞对过量营养应激的反应机制，并确定相关的生物标志物。
INS-1E 细胞在高葡萄糖、棕榈酸中单独或联合培养 24 小时，分别模拟糖毒性、脂毒性和糖脂毒性条件。进行生化和细胞实验以确认这些条件的建立。为了获得分子见解，使用 H-NMR 鉴定和定量大量代谢物。
高葡萄糖培养中未观察到细胞活力丧失，而单独或与高葡萄糖联合暴露于 FFA 会导致 ROS 水平升高、膜损伤、脂质积累和 DNA 双链断裂。使用 H-NMR 鉴定和定量了 49 种丰富的代谢物。在糖毒性和脂毒性条件下的化学计量两两分析，与糖脂毒性条件相比，失调代谢物存在部分重叠；然而，在糖脂毒性条件下，失调更为显著。
本研究平行比较了糖毒性、脂毒性和糖脂毒性条件，阐明了在糖尿病中起主要作用的代谢途径的差异。o-磷酸胆碱和 UDP-N-乙酰葡萄糖胺被鉴定为共同失调的代谢物，其比值被提议作为这些条件的潜在生物标志物。
