Christian Patry1,2, Kathrin Plotnicki1, Christian Betzen1,3, Alba Perez Ortiz4, Kirk L Pappan5, Simon C Satchell6, Peter W Mathieson7, Martina Bielaszewska8,9, Helge Karch8, Burkhard Tönshoff1, Neysan Rafat10,11,12. 1. Department of Pediatrics I, University Children's Hospital Heidelberg, 69120, Heidelberg, Germany. 2. Division of Cardiovascular Physiology, Institute of Physiology and Pathophysiology, University of Heidelberg, 69120, Heidelberg, Germany. 3. Division of Functional Genome Analysis, German Cancer Research Center (DKFZ), 69120, Heidelberg, Germany. 4. Department of Neonatology, University Children's Hospital Mannheim, University of Heidelberg, 68167, Mannheim, Germany. 5. Metabolon, Inc., 617 Davis Drive, Suite 400, Durham, NC, 27713, USA. 6. Learning and Research Southmead Hospital Bristol, University of Bristol, Bristol, BS8 1TH, UK. 7. The Principal's Office, University of Edinburgh, Edinburgh, EH8 9YL, UK. 8. Institute for Hygiene, University of Münster, 48149, Münster, Germany. 9. Reference Laboratory for E. coli and Shigella, National Public Health Institute, Srobarova 48, 10042, Prague, Czech Republic. 10. Department of Pediatrics I, University Children's Hospital Heidelberg, 69120, Heidelberg, Germany. Neysan.Rafat@umm.de. 11. Department of Neonatology, University Children's Hospital Mannheim, University of Heidelberg, 68167, Mannheim, Germany. Neysan.Rafat@umm.de. 12. Department of Pharmaceutical Sciences, Bahá'í Institute of Higher Education (BIHE), Teheran, Iran. Neysan.Rafat@umm.de.
Abstract
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.
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.
Authors: David C Nieman; R Andrew Shanely; Nicholas D Gillitt; Kirk L Pappan; Mary Ann Lila Journal: J Proteome Res Date: 2013-09-13 Impact factor: 4.466
Authors: S C Satchell; C H Tasman; A Singh; L Ni; J Geelen; C J von Ruhland; M J O'Hare; M A Saleem; L P van den Heuvel; P W Mathieson Journal: Kidney Int Date: 2006-05 Impact factor: 10.612