WZB117-Induced Glucose Hypometabolism Triggers Mitochondrial Dysfunction and Amyloidogenic Processing in an In Vitro Primary Forebrain Neuron Model

Background Alzheimer's disease (AD) is a progressive neurodegenerative disorder in which cerebral glucose hypometabolism represents one of the earliest and most consistent pathological abnormalities, often preceding classical amyloid and tau pathology. Despite strong clinical evidence, the causal contribution of impaired neuronal glucose utilization to AD-related cellular alterations remains incompletely understood. Objective This study aimed to determine whether selective inhibition of neuronal glucose transport induces AD-relevant cellular stress responses, including mitochondrial dysfunction and amyloidogenic processing, in a controlled primary neuronal system. Methods Primary forebrain neurons were established from embryonic chick brain and subjected to glucose hypometabolism using WZB117, a pharmacological inhibitor of facilitative glucose transporters. Neuronal viability and cytotoxicity were assessed by trypan blue exclusion and lactate dehydrogenase release assays. Mitochondrial membrane potential was evaluated using TMRE (tetramethylrhodamine ethyl ester) fluorescence, while amyloidogenic processing was quantified by measuring β-secretase (BACE1) activity and intracellular Aβ42 levels. β-hydroxybutyrate was employed to assess metabolic rescue. Results Inhibition of glucose transport resulted in a significant increase in neuronal cell death and cytotoxicity, accompanied by a pronounced reduction in mitochondrial membrane potential, indicating severe bioenergetic failure. Concurrently, amyloidogenic processing was markedly enhanced, as evidenced by elevated BACE1 activity and increased intracellular Aβ42 accumulation. Metabolic supplementation with β-hydroxybutyrate significantly attenuated neuronal death, restored mitochondrial function, and suppressed amyloidogenic alterations. Conclusions These findings demonstrate that acute neuronal glucose hypometabolism induces mitochondrial depolarization and increased amyloidogenic signaling in primary neurons, suggesting that metabolic stress may contribute to cellular processes associated with AD. The study identifies impaired neuronal bioenergetics as a potential upstream contributor to AD-related cellular pathology and supports further investigation of metabolism-focused strategies in AD research.