Introduction: Addiction to tramadol and pregabalin has escalated over a decade. Long-term tramadol use was linked to mitochondrial alteration in rodents’ brains. Also, pregabalin (PGB) abuse was proven to cause brain insults. Aim of the Work: The current study aimed to evaluate the impacts of pregabalin and tramadol on the cerebral cortex histological image and AMPK status in adult albino rats. Materials and Methods: thirty adult male albino rats were classified into three major groups: the control group, which obtained no medicine at any point throughout the study, the tramadol-treated group, which was administered a gradually increased dosage of tramadol, and the pregabalin-treated group, which obtained a gradually growing pregabalin dosage. The study ran for ninety days. Tramadol and pregabalin cytotoxicity were evaluated histologically and biochemically through assessing AMPKα1 & AMPKα2 mRNA expression and the apoptosis-related markers Bax & Bcl-2 by qRT-PCR. Results: Histological analysis in the tramadol and pregabalin groups showed aberrant disruption in cerebral cortical layers in addition to cellular apoptosis. Biochemical data showed that the neurotoxic effect of tramadol and pregabalin is mediated via oxidative stress, inflammation and AMPK expression in the brain of rats. Furthermore, there was a notable rise in the expression of the AMPKα1 and AMPKα2 genes. Conclusion: male albino rats were found to be neurotoxically affected by both tramadol and pregabalin. However, these impacts were less severe with Pregabalin. oxidative stress, inflammation, apoptosis, and the upregulation of AMPK expression are the mechanisms mediating the neurotoxic consequences caused by tramadol and pregabalin abuse. When these medications were misused, the brain's antioxidant defense enzyme activity was lowered, lipid peroxidation and 8-OHdG were elevated. Furthermore, both exhibited downregulation of anti-apoptotic proteins and increased the activation/expression of inflammation and apoptosis indicators, AMPKα1 & AMPKα2 in the rats' brains