Modulation of the AMPK/TFEB Axis by Ezetimibe Attenuates Neuroinflammatory, Oxidative Stress, and Neurotransmitter Dysregulation in Naloxone-precipitated Tramadol Withdrawal in Mice.

Tramadol withdrawal is associated with neuroinflammation, oxidative stress, and neurotransmitter imbalance, yet effective therapeutic strategies remain limited. Activation of the AMPK–TFEB (AMP-activated protein kinase-transcription factor EB) signaling axis enhances autophagy and cellular homeostasis and may mitigate withdrawal-associated neurotoxicity. To investigate whether ezetimibe attenuates naloxone-precipitated tramadol withdrawal in mice through modulation of the AMPK/TFEB pathway. Swiss albino mice received chronic tramadol exposure followed by naloxone to induce withdrawal. Ezetimibe (5 and 10 mg/kg, p.o.) was administered with or without the TFEB inhibitor eltrombopag. Behavioral outcomes (withdrawal severity score, jumping frequency, hyperalgesia), oxidative stress and inflammatory markers, neurotransmitter levels, and molecular docking interactions with TFEB were evaluated. Ezetimibe significantly reduced withdrawal severity, jumping frequency, hyperalgesia, lipid peroxidation, glutamate levels, and pro-inflammatory cytokines, while restoring antioxidant status, dopamine, and serotonin. Co-administration of eltrombopag attenuated these effects. Docking analysis revealed a stable interaction between ezetimibe and TFEB. Ezetimibe ameliorates tramadol-withdrawal-induced neurobehavioral and molecular alterations, most likely via AMPK-mediated activation of TFEB and enhancement of autophagy, highlighting its therapeutic potential in opioid withdrawal.