Livogrit ameliorates acetaldehyde-induced steatosis in HepG2 cells through modulation of lipogenesis and β-oxidation pathways

Frequent alcohol consumption leads to development of alcoholic liver steatosis. Onset of steatosis is caused by alcohol metabolite ‘acetaldehyde’ causing intracellular accumulation of lipid bodies due to abnormal metabolism. Livogrit is a tri-herbal Ayurveda formulation prepared using Boerhavia diffusa L., Phyllanthus niruri L., and Solanum nigrum L. plant extracts. The formulation has earlier proven effective against carbon tetrachloride-induced toxicity and non-alcoholic fatty acid induced liver steatosis. In present study, we evaluated efficacy of Livogrit in ameliorating ‘acetaldehyde’ induced steatosis and assessed its mode of action. HepG2 cells were exposed to varying concentrations of acetaldehyde and assessed for lipid body accumulation and cytosafety. An inhibition concentration of 50% for acetaldehyde was observed at 71.42 mM in HepG2 cells. 1 mM acetaldehyde was selected as a non-cytotoxic steatosis-stimulating dose. The acetaldehyde-stimulated HepG2 cells were used for studying the disease-reversing efficacy of Livogrit. Efficacy of the formulation was analyzed at cellular, biochemical, and molecular levels. Acetaldehyde stimulation caused an increase in intracellular lipid and triglyceride accumulation, release of ALT and AST enzymes, ROS generation, and decrease in extracellular release of glycerol molecules in HepG2 cells. Livogrit treatment of the stimulated-HepG2 cells showed a reversal of steatosis through normalization of observed intra- and extracellular parameters. At gene level, Livogrit-modulated the acetaldehyde-stimulated upregulation of SREBP1c, FAS, PLIN2, TNF-α, NF-kB, and LC3A genes associated with lipogenesis, inflammation, and autophagy. At protein level, the protective role of Livogrit was observed in acetaldehyde-stimulated HepG2 cells through modulation of TNF-α, PLIN2, and LC3 proteins. Livogrit provided further protection against steatosis through stimulation of fatty acid β-oxidation associated with SIRT1, CPT1A, and PPAR-α genes. In conclusion, Livogrit recovered HepG2 cells from alcoholic steatosis at biochemical and molecular levels through multifaceted modulation of lipogenesis, inflammation, autophagy, and β-oxidation. Hence, Livogrit was found to be a potential candidate for the amelioration of alcohol-induced steatosis.