INTRODUCTION: Shiga toxin 2a (Stx2a) induces hemolytic uremic syndrome (STEC HUS) by targeting glomerular endothelial cells (GEC). OBJECTIVES: 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. METHODS: 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. RESULTS: 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. CONCLUSION: Stimulation of ciGEnC with TNFα + Stx2a is associated with profound metabolic changes indicative of increased inflammation, oxidative stress and energy turnover.

Department of Neonatology University Children's Hospital Mannheim University of Heidelberg 68167 Mannheim Germany

Department of Pediatrics 1 University Children's Hospital Heidelberg 69120 Heidelberg Germany

Department of Pharmaceutical Sciences Bahá'í Institute of Higher Education Teheran Iran

Division of Cardiovascular Physiology Institute of Physiology and Pathophysiology University of Heidelberg 69120 Heidelberg Germany

Division of Functional Genome Analysis German Cancer Research Center 69120 Heidelberg Germany

Institute for Hygiene University of Münster 48149 Münster Germany

Learning and Research Southmead Hospital Bristol University of Bristol Bristol BS8 1TH UK

Metabolon Inc 617 Davis Drive Suite 400 Durham NC 27713 USA

Reference Laboratory for E coli and Shigella National Public Health Institute Srobarova 48 10042 Prague Czech Republic

The Principal's Office University of Edinburgh Edinburgh EH8 9YL UK

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