Short‐Chain Fatty Acid Mediated Protection Against Oxidative Stress by Lacticaseibacillus paracasei in HepG2 Cells

Metabolic dysfunction associated steatohepatitis (MASH) (formerly nonalcoholic steatohepatitis, NASH) is a progressive manifestation of metabolic dysfunction‐associated steatotic liver disease, strongly linked to oxidative stress and gut–liver axis dysregulation. Targeting microbial metabolites such as short‐chain fatty acids (SCFAs) through dietary interventions represents a promising avenue for hepatoprotection. In this study, we developed a probiotic‐fortified grape juice incorporating Lacticaseibacillus paracasei and its secondary metabolites and investigated its stability and potential protective effects in a cellular model of NASH. L. paracasei was cultured under optimized conditions, and its metabolite extract was characterized by Fourier‐transform infrared spectroscopy, which confirmed the presence of SCFAs and other bioactive functional groups. The fortified juice was evaluated for physicochemical attributes, microbial stability, and probiotic viability over 15 days of refrigerated storage. The formulation retained acceptable sensory quality while maintaining probiotic counts above 8 log CFU/mL until Day 15, with no evidence of spoilage. To assess hepatoprotective potential, HepG2 cells exposed to palmitic acid were used to simulate NASH‐like oxidative stress. Treatment with the fortified juice extract reduced intracellular ROS levels by 48.2 ± 3.5 % ( p < 0.01) compared with untreated controls while preserving cell viability (< 20% cytotoxicity, mean ± SD, n = 3). These findings suggest that the combination of live probiotics and their postbiotic metabolites within a fruit‐based delivery system can attenuate oxidative stress and potentially restore redox homeostasis in hepatocytes. This work highlights the translational potential of leveraging L. paracasei ‐derived SCFAs as part of precision nutrition strategies for MASH management. Beyond liver health, such formulations may represent a broader platform for microbiota‐targeted dietary interventions aimed at mitigating oxidative damage and supporting gut–liver axis integrity.