Tamuli Roktima, Mellick George D, Schirra Horst Joachim, Feng Yunjiang
Institute for Biomedicine and Glycomics, Griffith University, Nathan, QLD 4111, Australia.
School of Environment and Science, Griffith University, Nathan, QLD 4111, Australia.
Molecules. 2025 Aug 12;30(16):3352. doi: 10.3390/molecules30163352.
This study used NMR-based metabolomics to investigate the mode of action (MoA) of 6-hydroxydopamine (6-OHDA) toxicity in the SH-SY5Y neuroblastoma cell model. 6-OHDA, a structural analogue of dopamine, has been used to create a Parkinson's disease model since 1968. Its selective uptake via catecholaminergic transporters leads to intracellular oxidative stress and mitochondrial dysfunction. SH-SY5Y cells were treated with 6-OHDA at its IC concentration of 60 μM, and samples of treated and untreated groups were collected after 24 h. The endo metabolome was extracted using a methanol-water mixture, while the exo metabolome was represented by the culture media. Further, endo- and exo metabolomes of treated and untreated cells were analysed for metabolic changes. Our results demonstrated significantly high levels of glutathione, acetate, propionate, and NAD, which are oxidative stress markers, enhanced due to ROS production in the system. In addition, alteration of myoinositol, taurine, and o-phosphocholine could be due to oxidative stress-induced membrane potential disturbance. Mitochondrial complex I inhibition causes electron transport chain (ETC) dysfunction. Changes in key metabolites of glycolysis and energy metabolism, such as glucose, pyruvate, lactate, creatine, creatine phosphate, glycine, and methionine, respectively, demonstrated ETC dysfunction. We also identified changes in amino acids such as glutamine, glutamate, and proline, followed by nucleotide metabolism such as uridine and uridine monophosphate levels, which were decreased in the treated group.
本研究采用基于核磁共振的代谢组学方法，在SH-SY5Y神经母细胞瘤细胞模型中研究6-羟基多巴胺（6-OHDA）毒性的作用模式（MoA）。6-OHDA是多巴胺的结构类似物，自1968年以来一直用于建立帕金森病模型。其通过儿茶酚胺能转运体的选择性摄取会导致细胞内氧化应激和线粒体功能障碍。用60μM的IC浓度的6-OHDA处理SH-SY5Y细胞，24小时后收集处理组和未处理组的样本。内源性代谢组用甲醇-水混合物提取，而外源性代谢组由培养基代表。此外，分析处理组和未处理组细胞的内源性和外源性代谢组的代谢变化。我们的结果表明，作为氧化应激标志物的谷胱甘肽、乙酸盐、丙酸盐和NAD水平显著升高，这是由于系统中活性氧的产生所致。此外，肌醇、牛磺酸和磷酸胆碱的改变可能是由于氧化应激诱导的膜电位紊乱。线粒体复合体I的抑制导致电子传递链（ETC）功能障碍。糖酵解和能量代谢的关键代谢物，如葡萄糖、丙酮酸、乳酸、肌酸、磷酸肌酸、甘氨酸和蛋氨酸的变化，分别表明ETC功能障碍。我们还发现谷氨酰胺、谷氨酸和脯氨酸等氨基酸发生了变化，随后核苷酸代谢如尿苷和单磷酸尿苷水平也发生了变化，这些在处理组中均有所下降。
