Investigating the epigenetic modulating potential of gallic acid during ethanol-induced toxicity: In silico and in vitro approaches.

Alcohol-related disorders contribute significantly to morbidity, mortality, and economic burdens worldwide. Excessive alcohol consumption affects almost all organs, particularly liver, intestine, and kidney, by generating excessive free radicals during metabolism, which disrupts cellular epigenetic homeostasis and contributes to the disease onset. One of the effective therapeutic strategy for alcohol-induced toxicity involves the use of immunosuppressants and corticosteroids, but with numerous side effects. The exploration of cost-effective, low-toxicity natural products for alcohol-associated liver disease is of growing interest. Moreover, the mechanistic relationship between oxidative stress-induced epigenetic dysregulation and ethanol-induced toxicity remains unexplored. Based on the above concepts, we investigated the effect of gallic acid (GA), a phytochemical known for its potent antioxidant activity against ethanol-induced hepatotoxicity. WRL 68 cells were treated with ethanol and GA under two different conditions: acute (24 h) and chronic (14 d). GA (30 μM) protected cells from oxidative stress by maintaining the levels of SOD, CAT, GSH, and TBARS in the ethanol-treated group. Under acute exposure to ethanol, GA significantly downregulated miR_21_5p, miR_17_5p, HAT, and HDM, and upregulated miR_199a_5p, miR_129_5p, miR_26b_5p, HDACs, and HMTs expression. GA preserved DNA integrity and prevented the cells from undergoing apoptosis by suppressing the expression of apoptotic genes (Cas 3, Cas 9, and Bax). During chronic exposure to ethanol, GA restored cellular homeostasis by maintaining the epigenetic balance of the cells and attenuated EMT progression in WRL 68 cells. Based on our findings, we suggest that GA can act as an antioxidant and protect cells from epigenetic dyshomeostasis during ethanol exposure. However, extensive analyses, such as protein expression profiling and ChIP assays, are required to understand the mechanistic insights of GA in maintaining the epigenetic balance of cells under ethanol toxicity.