Metabolic Flux Analysis

Metabolomics reflects the static abundance ratio of metabolites, and the increase of a single metabolite may be due to either the active synthetic pathway or the inhibition of the depletion pathway. Therefore, metabonomics is often inadequate to explain all the problems in the study of specific metabolic pathways and metabolic networks.

As a complement to metabonomics, Metabolic Flux analysis uses stable isotope to label specific molecules and track their metabolism in an organism, thus providing dynamic information on the Metabolic pathways of metabolites.

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Detect Content

• Glycolysis
• TCA cycle
• Pentose phosphate pathways

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Sample Requirements

• Cells in the sample≥ 4×10⁶
• Store in liquid nitrogen or -80°C
• Dry ice shipping

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Analysis Process

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Case Analysis

Regulate metabolic pathways in vivo to inhibit the occurrence of acute and chronic diseases Gout and Pseudo-gout-related Crystals Promote GLUT1-mediated Glycolysis that Governs NLRP3 and Interleukin-1β Activation on Macrophages

journal: Annals of the rheumatic diseases       Impact factor: 16.102 Published date: July 2019       Published by: University of Paris, France

Analysis Purpose

Deciphering the role of glycolysis and oxidative phosphorylation in IL-1β-induced microcrystal responses.

Analysis Method

Macrophages were stimulated with monosodium urate (MSU) and calcium pyrophosphate (CPP) crystals and subjected to in vitro metabolomics and real-time extracellular flux analysis to observe the metabolic phenotype of macrophages. Using f-deoxyglucose positron emission tomography, glucose uptake experiments, and molecular biological methods that inhibit glucose invertase (Glut1) , evaluation of the effects of aerobic glycolysis on IL-1 β production in vitro and in vivo.

Analysis Results

We observed that MSU and CPP crystals led to metabolic reorganization of macrophages into the aerobic glycolytic pathway, which is caused by increased glucose uptake and expression of GLUT1 plasma membrane in macrophages. Furthermore, neutrophils isolated from human synovial fluid during gout flares express GLUT1 more frequently than neutrophils isolated from blood. Both 2-deoxyglucose or GLUT1 inhibited NLRP3 activation and IL-1β production, thereby inhibiting microcrystalline inflammation.

GLUT1 and glycolysis regulate NLRP3 inflammasome activation and IL-1β production induced by MSU and m-CPPD crystals.

MSU and m-CPPD crystals induce metabolic changes in the glycolysis pathway and the TCA cycle

Conclusion

GLUT1-mediated glucose uptake plays an important role in MSU- and CPP crystal-induced IL-1β protein depletion responses. These findings open new avenues for the treatment of arthritis in both acute and chronic patients.

Reference

Renaudin Felix,Orliaguet Lucie,Castelli Florence et al. Gout and pseudo-gout-related crystals promote GLUT1-mediated glycolysis that governs NLRP3 and interleukin-1β activation on macrophages.[J] .Ann. Rheum. Dis., 2020