13 C-Metabolic flux analysis detected a hyperoxemia-induced reduction of tricarboxylic acid cycle metabolism in granulocytes during two models of porcine acute subdural hematoma and hemorrhagic shock.

Autor: Wolfschmitt EM; Institute for Anesthesiological Pathophysiology and Process Engineering, University Hospital Ulm, Ulm, Germany., Vogt JA; Institute for Anesthesiological Pathophysiology and Process Engineering, University Hospital Ulm, Ulm, Germany., Hogg M; Institute for Anesthesiological Pathophysiology and Process Engineering, University Hospital Ulm, Ulm, Germany., Wachter U; Institute for Anesthesiological Pathophysiology and Process Engineering, University Hospital Ulm, Ulm, Germany., Stadler N; Institute for Anesthesiological Pathophysiology and Process Engineering, University Hospital Ulm, Ulm, Germany., Kapapa T; Clinic for Neurosurgery, University Hospital Ulm, Ulm, Germany., Datzmann T; Clinic for Anesthesia and Intensive Care, University Hospital Ulm, Ulm, Germany., Messerer DAC; Institute for Anesthesiological Pathophysiology and Process Engineering, University Hospital Ulm, Ulm, Germany.; Institute for Transfusion Medicine, University Hospital Ulm, Ulm, Germany., Hoffmann A; Institute for Anesthesiological Pathophysiology and Process Engineering, University Hospital Ulm, Ulm, Germany., Gröger M; Institute for Anesthesiological Pathophysiology and Process Engineering, University Hospital Ulm, Ulm, Germany., Münz F; Institute for Anesthesiological Pathophysiology and Process Engineering, University Hospital Ulm, Ulm, Germany.; Clinic for Anesthesia and Intensive Care, University Hospital Ulm, Ulm, Germany., Mathieu R; Clinic for Neurosurgery, Bundeswehrkrankenhaus, Ulm, Germany., Mayer S; Clinic for Neurosurgery, Bundeswehrkrankenhaus, Ulm, Germany., Merz T; Institute for Anesthesiological Pathophysiology and Process Engineering, University Hospital Ulm, Ulm, Germany.; Clinic for Anesthesia and Intensive Care, University Hospital Ulm, Ulm, Germany., Asfar P; Département de Médecine Intensive - Réanimation et Médecine Hyperbare, Centre Hospitalier Universitaire, Angers, France., Calzia E; Institute for Anesthesiological Pathophysiology and Process Engineering, University Hospital Ulm, Ulm, Germany., Radermacher P; Institute for Anesthesiological Pathophysiology and Process Engineering, University Hospital Ulm, Ulm, Germany., Zink F; Institute for Anesthesiological Pathophysiology and Process Engineering, University Hospital Ulm, Ulm, Germany.
Jazyk: angličtina
Zdroj: Frontiers in immunology [Front Immunol] 2024 Jan 09; Vol. 14, pp. 1319986. Date of Electronic Publication: 2024 Jan 09 (Print Publication: 2023).
DOI: 10.3389/fimmu.2023.1319986
Abstrakt: Introduction: Supplementation with increased inspired oxygen fractions has been suggested to alleviate the harmful effects of tissue hypoxia during hemorrhagic shock (HS) and traumatic brain injury. However, the utility of therapeutic hyperoxia in critical care is disputed to this day as controversial evidence is available regarding its efficacy. Furthermore, in contrast to its hypoxic counterpart, the effect of hyperoxia on the metabolism of circulating immune cells remains ambiguous. Both stimulating and detrimental effects are possible; the former by providing necessary oxygen supply, the latter by generation of excessive amounts of reactive oxygen species (ROS). To uncover the potential impact of increased oxygen fractions on circulating immune cells during intensive care, we have performed a 13 C-metabolic flux analysis (MFA) on PBMCs and granulocytes isolated from two long-term, resuscitated models of combined acute subdural hematoma (ASDH) and HS in pigs with and without cardiovascular comorbidity.
Methods: Swine underwent resuscitation after 2 h of ASDH and HS up to a maximum of 48 h after HS. Animals received normoxemia (P a O 2 = 80 - 120 mmHg) or targeted hyperoxemia (P a O 2 = 200 - 250 mmHg for 24 h after treatment initiation, thereafter P a O 2 as in the control group). Blood was drawn at time points T1 = after instrumentation, T2 = 24 h post ASDH and HS, and T3 = 48 h post ASDH and HS. PBMCs and granulocytes were isolated from whole blood to perform electron spin resonance spectroscopy, high resolution respirometry and 13 C-MFA. For the latter, we utilized a parallel tracer approach with 1,2- 13 C 2 glucose, U- 13 C glucose, and U- 13 C glutamine, which covered essential pathways of glucose and glutamine metabolism and supplied redundant data for robust Bayesian estimation. Gas chromatography-mass spectrometry further provided multiple fragments of metabolites which yielded additional labeling information. We obtained precise estimations of the fluxes, their joint credibility intervals, and their relations, and characterized common metabolic patterns with principal component analysis (PCA).
Results: 13 C-MFA indicated a hyperoxia-mediated reduction in tricarboxylic acid (TCA) cycle activity in circulating granulocytes which encompassed fluxes of glutamine uptake, TCA cycle, and oxaloacetate/aspartate supply for biosynthetic processes. We further detected elevated superoxide levels in the swine strain characterized by a hypercholesterolemic phenotype. PCA revealed cell type-specific behavioral patterns of metabolic adaptation in response to ASDH and HS that acted irrespective of swine strains or treatment group.
Conclusion: In a model of resuscitated porcine ASDH and HS, we saw that ventilation with increased inspiratory O 2 concentrations (P a O 2 = 200 - 250 mmHg for 24 h after treatment initiation) did not impact mitochondrial respiration of PBMCs or granulocytes. However, Bayesian 13 C-MFA results indicated a reduction in TCA cycle activity in granulocytes compared to cells exposed to normoxemia in the same time period. This change in metabolism did not seem to affect granulocytes' ability to perform phagocytosis or produce superoxide radicals.
Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
(Copyright © 2024 Wolfschmitt, Vogt, Hogg, Wachter, Stadler, Kapapa, Datzmann, Messerer, Hoffmann, Gröger, Münz, Mathieu, Mayer, Merz, Asfar, Calzia, Radermacher and Zink.)
Databáze: MEDLINE
Externí odkaz: