Vanillic Acid Mitigates Alcohol-Induced Oxidative Stress by Modulating Lipid Peroxidation, Nitric Oxide Levels, and Antioxidant Enzyme Activity in Zebrafish Larvae

Abstract

Alcohol consumption is a major contributor to oxidative stress-related diseases, including liver damage and neurotoxicity. Vanillic acid (VA), a natural phenolic compound, possesses strong antioxidant properties that may mitigate oxidative damage. This study evaluates the protective effects of VA against alcohol-induced oxidative stress in zebrafish larvae. Zebrafish were exposed to 1% ETOH for 24 hours to induce oxidative stress, followed by treatment with VA at concentrations of 50 μg/mL and 100 μg/mL for another 24 hours. The impact of VA on oxidative stress biomarkers was assessed by measuring lipid peroxidation, nitric oxide production, and antioxidant enzyme activities, including superoxide dismutase, catalase, glutathione, and glutathione S-transferase. Additionally, acridine orange staining was used to evaluate apoptosis, while reactive oxygen species levels were quantified using dichlorofluorescin diacetate and lipid peroxidation was analyzed using diphenyl-1-pyrenylphosphine fluorescence assays. Results demonstrated a significant increase in oxidative stress markers following 1% ETOH exposure, characterized by elevated ROS levels, MDA accumulation, and reduced antioxidant enzyme activities. However, VA treatment effectively counteracted these effects, with 100 μg/mL showing the
highest efficacy in restoring antioxidant defenses, reducing lipid peroxidation, and inhibiting NO overproduction. Furthermore, fluorescence-based assays confirmed the reduction of apoptosis and oxidative damage in VA-treated zebrafish larvae. These findings suggest that VA mitigates alcohol-induced oxidative stress by modulating key biochemical pathways, thereby offering potential therapeutic benefits against alcohol-related toxicity. Future studies should focus on elucidating the molecular mechanisms underlying VA’s protective effects.