| Summary: | Alcoholic liver fatty disease (ALFD) is caused by excessive and chronic alcohol consumption. Alcohol consumption causes an imbalance in the intestinal microflora, leading to liver disease induced by the excessive release of endotoxins into the hepatic portal vein. Therefore, research on the intestinal microflora to identify treatments for ALFD is increasing. In this study, the protective effects of lactic acid bacteria (LAB) strains, including <i>Levilactobacillus brevis, Limosilactobacillus reuteri</i>, and <i>Limosilactobacillus fermentum</i>, were evaluated in ethanol-induced HepG2 cells. Among the evaluated LAB, nine strains increased aldehyde dehydrogenase (ALDH) levels and downregulated lipid peroxidation and liver transferase in the ethanol-induced HepG2 cells. Moreover, <i>L. brevis</i> MG5280 and MG5311, <i>L. reuteri</i> MG5458, and <i>L. fermentum</i> MG4237 and MG4294 protected against ethanol-induced HepG2 cell damage by regulating <i>CYP2E1</i>, antioxidant enzymes (<i>SOD</i>, <i>CAT</i>, and <i>GPX</i>), lipid synthesis factors (<i>SREBP1C</i> and <i>FAS</i>), and lipid oxidation factors (<i>PPARα</i>, <i>ACO</i>, and <i>CPT-1</i>). Moreover, five LAB were confirmed to be safe probiotics based on antibiotic susceptibility and hemolysis assays; their stability and adhesion ability in the gastrointestinal tract were also established. In conclusion, <i>L. brevis</i> MG5280 and MG5311, <i>L. reuteri</i> MG5458, and <i>L. fermentum</i> MG4237 and MG4294 may be useful as new probiotic candidates for ALFD prevention.
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