Department of Pediatrics I, University Children's Hospital Heidelberg, 69120, Heidelberg, Germany.
Division of Cardiovascular Physiology, Institute of Physiology and Pathophysiology, University of Heidelberg, 69120, Heidelberg, Germany.
Metabolomics. 2019 Oct 1;15(10):131. doi: 10.1007/s11306-019-1594-2.
Shiga toxin 2a (Stx2a) induces hemolytic uremic syndrome (STEC HUS) by targeting glomerular endothelial cells (GEC).
We investigated in a metabolomic analysis the response of a conditionally immortalized, stable glomerular endothelial cell line (ciGEnC) to Stx2a stimulation as a cell culture model for STEC HUS.
CiGEnC were treated with tumor necrosis factor-(TNF)α, Stx2a or sequentially with TNFα and Stx2a. We performed a metabolomic high-throughput screening by lipid- or gas chromatography and subsequent mass spectrometry. Metabolite fold changes in stimulated ciGEnC compared to untreated cells were calculated.
320 metabolites were identified and investigated. In response to TNFα + Stx2a, there was a predominant increase in intracellular free fatty acids and amino acids. Furthermore, lipid- and protein derived pro-inflammatory mediators, oxidative stress and an augmented intracellular energy turnover were increased in ciGEnC. Levels of most biochemicals related to carbohydrate metabolism remained unchanged.
Stimulation of ciGEnC with TNFα + Stx2a is associated with profound metabolic changes indicative of increased inflammation, oxidative stress and energy turnover.
志贺毒素 2a（Stx2a）通过靶向肾小球内皮细胞（GEC）诱导溶血性尿毒症综合征（STEC HUS）。
我们通过代谢组学分析研究了条件永生化稳定肾小球内皮细胞系（ciGEnC）对 Stx2a 刺激的反应，作为 STEC HUS 的细胞培养模型。
用肿瘤坏死因子-（TNF）α、Stx2a 或 TNFα 和 Stx2a 序贯处理 ciGEnC。我们通过脂质或气相色谱和随后的质谱进行了代谢组学高通量筛选。与未处理的细胞相比，刺激的 ciGEnC 中的代谢物倍数变化被计算出来。
鉴定并研究了 320 种代谢物。在 TNFα+Stx2a 反应中，细胞内游离脂肪酸和氨基酸明显增加。此外，ciGEnC 中的炎症介质、氧化应激和细胞内能量周转增加。与碳水化合物代谢相关的大多数生化物质的水平保持不变。
ciGEnC 用 TNFα+Stx2a 刺激与炎症、氧化应激和能量周转增加相关的深刻代谢变化相关。
