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 Adenosine 5’-monophosphatase-activated protein kinase (AMPK) status in adult albino rats. Materials and Methods: thirty adult male albino rats were classified into three major groups ten rats each: Control group, which obtained no medicine at any point throughout the study, Tramadol-treated group, which was administered a gradually increased dosage of tramadol starting with 30 mg/kg/day (1/5th LD50) for 30 continuous days, then 60 mg/kg/day for another 30 days then 120 mg/kg/day in the last 30 days of the study, and Pregabalin-treated group, which obtained a gradually growing (PGB) dosage starting with 300 mg/day 3 times/week till reaching dependence level within a month then 3400 mg for additional 2 months the study ran for ninety days. Tramadol and pregabalin cytotoxicity were evaluated histologically and biochemically through assessing AMPKα1 and AMPKα2 mRNA expression and the apoptosis-related markers Bax and 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.