Aims. Acute consumption of sweeteners has become increasingly common in modern diets. However, emerging research suggests that the impact of sweeteners on the hypothalamus, involved in regulating food intake [CA1] and metabolism, may be deleterious. Several studies have raised concerns about their potential effects on brain cellular metabolism (Kohno D et al.,2016), suggesting a mismatch between detection of false sugar and the energy value actually detected. This study aimed to investigate the dose-dependent impact of sweeteners on these energetic processes. Methods. Using the hypothalamic cell line GT1-7, we assessed ATP levels in response to varying concentrations of two commonly used sweeteners Sucralose (0, 0.01mM, 0.1mM, 1mM and 5mM) and AceK (0, 0.03mM, 0.3mM, 3mM and 15mM) for 30 min concomitantly with stimulation with 20 mM glucose. ATP concentrations were measured using a luciferin/luciferase bioluminescent assay. For mitochondrial respiration, oxygen consumption was measured (Oxygraph-2k respirometer; Oroboros Instruments) using 2-3 millions GT1-7 cells transferred in mitochondrial respiration medium (MIR05) in the oxygraph chamber. Respiratory capacities were measured in the basal state and then stimulated by adding 20 mM succinate (substrate-driven respiration) before or after increasing concentrations of sweeteners (10-3, 10-2 or 1 mM of sucralose or AceK). Results. Higher doses of sweeteners significantly impair both ATP production and mitochondrial respiration, highlighting that these sweeteners alter cellular oxidative metabolism. Conclusion. These findings underscore the importance of further research to elucidate the mechanisms underlying the effects of non-nutritive sweeteners on the energy capacities of neuronal cells, albeit with sufficient glucose