Nonalcoholic fatty liver disease (NAFLD) is currently one of the most common liver diseases worldwide. Lifestyle modifications through the diet are the mainstay of treatment. Auricularia nigricans is a popular edible mushroom known to possess medicinal properties. Gas chromatography-mass spectrometry and liquid chromatography-tandem mass spectrometry analysis indicated that linoleic acid ethyl ester, butyl 9,12-octadecadienoate, 9,12-octadecadienoic acid, ergosta-5,7,22-trien-3-ol, 2(3,4-dihydroxyphenyl)-7-hydroxy-5-benzene propanoic acid, and 3,30-di-0-methyl ellagic acid were present in the A. nigricans ethyl acetate (EA) fraction. The cytotoxicity assay showed that the EA fraction was noncytotoxic to HepG2 cells at concentrations < 100 μg/mL. In the antihepatic steatosis assay, 50 μg/mL of EA fraction caused a decline in absorbance to 0.20 ± 0.02 compared to palmitic acid (PA)-induced cells (0.24 ± 0.02). Furthermore, cells treated with 50 μg/mL and 25 μg/mL of EA fraction contributed an approximately 1.12-fold and 1.08-fold decrease in lipid accumulation compared to PA-induced cells. Coincubation with PA and 25 μg/mL of EA fraction decreased levels of tumor necrosis factor-α, interleukin (IL)-6, IL-8, and monocyte chemoattractant protein-1 to 140.48 ± 8.12, 91.16 ± 2.40, 184.00 ± 22.68, and 935.88 ± 39.36 pg/mL compared to PA-induced cells. The presence of the EA fraction also suppressed the stress-activated protein kinase/Jun amino-terminal kinase, p-38 mitogen-activated protein kinase, nuclear factor-κB, and signal transducer and activator of transcription 3 signaling pathways. In conclusion, these findings suggest that the A. nigricans EA fraction demonstrates antisteatotic effects involving antioxidant capacity, hypolipidemic effects, and anti-inflammatory capacity in the PA-induced NAFLD pathological cell model.
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