Evaluation of Rohypnol-Induced Acute Neurotoxicity in Male Wistar Rats

Rohypnol (Flunitrazepam), a benzodiazepine, is commonly used as a pre- anaesthetic and for insomnia treatment. However, it is also frequently abused for its sedative and amnestic effects, raising concerns regarding its neurotoxic potential. This study aimed to evaluate the acute neurotoxic effects of Rohypnol on cognitive function, cholinergic activity, and oxidative stress in the brains of male Wistar rats. Twenty male Wistar rats (180–200 g) were divided into two groups: control (Group A) and Rohypnol-treated (Group B). Rohypnol was administered at a dose of 0.1 mg/kg orally for 7 days. Behavioral testing included the T-maze to assess spatial memory, the Y-maze for spontaneous alternation, and the Light-Dark box test for anxiety- like behavior. Biochemical assays were performed to measure acetylcholinesterase (AChE) activity, and oxidative stress markers, including malondialdehyde (MDA) levels, superoxide dismutase (SOD), glutathione peroxidase (GST), glutathione (GSH), and catalase activity in brain homogenates. Rats treated with Rohypnol showed a significant reduction in spontaneous alternation in the Y-maze, suggesting impairments in spatial memory (p < 0.05). Rohypnol-exposed rats also spent significantly more time in the dark compartment of the Light-Dark box test, indicating increased anxiety-like behavior (p < 0.05). Biochemically, Rohypnol administration led to a significant increase in AChE activity and MDA levels, indicating disrupted cholinergic signaling and neuronal oxidative damage (p < 0.05). Additionally, oxidative stress markers including SOD, GSH, GST, and catalase were significantly elevated in the brains of Rohypnol-treated rats (p < 0.05), suggesting a compensatory response to oxidative damage. Acute exposure to Rohypnol induces significant neurotoxic effects in male Wistar rats, as evidenced by impaired cognitive performance, increased anxiety-like behavior, and biochemical markers of oxidative stress and cholinergic dysfunction. These findings highlight the potential neurotoxic effects of Rohypnol, particularly in cases of misuse or overdose, and underscore the need for further research on its long-term impact on brain function and health.