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Here, we show that <i>Lactiplantibacillus plantarum</i> LP158 (LP158), <i>Lactobacillus helveticus</i> HY7804 (HY7804), and <i>Lacticaseibacillus paracasei</i> LPC226 (LPC226) isolated from raw milk alleviate non-alcoholic fatty acid disease (NAFLD) in a C57BL/6 mouse model. Lactic acid bacteria (LAB) were screened for their ability to inhibit fatty acid accumulation in palmitic acid (PA)-treated HepG2 cells, and three strains were selected based on the results. We also investigated hemolytic activity and antibiotic resistance of the three strains. LP158, HY7804, and LPC226 suppressed expression of mRNA encoding genes related to lipogenesis, and increased expression of genes related to β-oxidation, in a PA-induced HepG2 cell model. Moreover, when LP158, HY7804, and LPC226 were administered at 10⁹ CFU/kg/day for 8 weeks to mice with dietary-induced NAFLD, they all modulated blood biochemistry markers and reduced steatosis in liver tissue. Also, all three strains significantly reduced expression of mRNA encoding lipogenesis genes (<i>Fasn</i>, <i>Acaca</i>, and <i>Srebp-1c</i>) and inflammatory factors (<i>Tnfα</i> and <i>Ccl-2</i>) and fibrosis factors, and increased expression of a β-oxidation gene (<i>Acox1</i>) in the liver. In particular, HY7804 showed the strongest effects both in vitro and in vivo. Therefore, HY7804, LP158, and LPC226 can be proposed as potential supplements that can improve NAFLD through anti-steatosis, anti-inflammatory, and anti-fibrotic effects.