Ambroxol's Role in PTZ-Induced Anxiety-Like Conditions in Mice: Targeting Neuroinflammation and Oxidative Stress via In Vivo and In Silico Approaches

Abstract

Anxiety disorders are major neuropsychiatric conditions linked to oxidative stress, neuroinflammation, and dysregulated neurotransmission. Current pharmacotherapies, such as benzodiazepines, offer symptomatic relief but are limited by tolerance and sedation. Ambroxol, a clinically established mucolytic agent, has demonstrated neuroprotective, anti-inflammatory, and antioxidant activities, suggesting potential therapeutic repurposing for neuropsychiatric disorders. The present study evaluated the anxiolytic and neuroprotective effects of ambroxol in a pentylenetetrazol (PTZ)-induced anxiety model in mice, supported by molecular docking and molecular dynamics analyses, showing moderate but significant interactions. Mice received ambroxol (60 mg/kg, p.o.) for 21 days, while diazepam (2 mg/kg, i.p.) served as the reference control for 14 days. Behavioral assessments were performed using the elevated plus maze, open field, light-dark box, and hole board tests. Biochemical analysis of prefrontal cortical tissue was conducted to evaluate oxidative stress and pro-inflammatory markers (MDA, SOD, CAT, GSH, TNF-alpha, IL-1 beta, NF-kappa B). Ambroxol significantly improved exploratory and anxiety-related behaviors, restored antioxidant enzyme activities, and suppressed cytokine levels in the prefrontal cortex. Docking and simulation studies revealed stable, biologically relevant interactions of ambroxol with TNF-alpha, IL-1 beta, and NF-kappa B, suggesting modulation of neuroimmune signaling. Ambroxol exhibited anxiolytic efficacy comparable to diazepam, with no apparent sedative effects observed behaviorally, although direct assessment of sedation (e.g., rotarod test) was not performed. Ambroxol demonstrated promising multi-target neuroprotective potential by concurrently attenuating oxidative and inflammatory cascades, supporting its further exploration as a safe, repurposed therapeutic candidate for anxiety disorders.