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Updated: Aug 25, 2026
Screening Assay for Oxidative Stress in a Feline Astrocyte Cell Line, G355-5
Published on: July 13, 2011
Glucocerebroside inhibits NADPH oxidase activation in cell-free system
Patryk Moskwa1, Anita Palicz, Marie-Hélène Paclet
1Department of Physiology, Semmelweis University, Budapest, Hungary.
Abstract:
We reported earlier that monocytes and macrophages from patients with type I Gaucher disease have a decreased capacity to generate superoxide anion (O(2)(-)) on stimulation with opsonized S. aureus or formyl-methionyl-leucyl-phenylalanine. In this study, various forms of the cell-free assay system were used to probe the hypothesis that glucocerebroside (GC) accumulating in Gaucher patients' phagocytes may interfere with the activation of NADPH oxidase. Xanthine/xanthine oxidase assay was applied to explore the possibility that GC may scavenge O(2)(-). We found that addition of GC to the crude, semirecombinant or fully purified cell-free systems inhibited activation of NADPH oxidase in a concentration-dependent manner. The inhibitory effect of GC could be overcome by increased concentrations of p47(phox) and p67(phox). In contrast, O(2)(-) generation was not decreased by GC added to the assembled, catalytically active enzyme complex. In the xanthine/xanthine oxidase system, GC had no effect on the generation of O(2)(-). These data indicate that assembly of the respiratory burst oxidase of phagocytic cells may be a possible target of the pathologic actions of GC.
Insights
Glucocerebroside (GC) accumulation in Gaucher disease impairs phagocyte NADPH oxidase assembly. This dysfunction in superoxide anion (O(2)(-)) generation is linked to GC
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Monocytes and macrophages in Gaucher disease exhibit reduced superoxide anion (O(2)(-)) generation.
- Glucocerebroside (GC) accumulates in Gaucher patients' phagocytes.
Purpose of the Study:
- To investigate if accumulating glucocerebroside (GC) interferes with NADPH oxidase activation in Gaucher disease.
- To determine the mechanism by which GC affects O(2)(-) generation.
Main Methods:
- Utilized cell-free assay systems (crude, semirecombinant, purified) to study NADPH oxidase activation.
- Employed xanthine/xanthine oxidase assay to assess GC's potential to scavenge O(2)(-).
- Investigated the effect of varying concentrations of p47(phox) and p67(phox) on GC's inhibitory action.
Main Results:
- Glucocerebroside (GC) inhibited NADPH oxidase activation in a concentration-dependent manner in cell-free systems.
- The inhibitory effect of GC was reversed by increasing concentrations of p47(phox) and p67(phox).
- GC did not affect O(2)(-) generation when added to the pre-assembled, active enzyme complex or in the xanthine/xanthine oxidase system.
Conclusions:
- Glucocerebroside (GC) accumulation interferes with the assembly of the respiratory burst oxidase.
- The assembly process of NADPH oxidase is a potential target for the pathological effects of GC in Gaucher disease.
- This finding provides insight into the cellular dysfunction observed in Gaucher disease phagocytes.
