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Acetaminophen (APAP) or paracetamol is an analgesics and antipyretic drug that can cause hepatotoxicity at an overdosage or long-term use. Paracetamol-induced hepatotoxicity is a public concern worldwide. The present study was aimed to investigate the effects of probiotics and synbiotics on the prevention of paracetamol-induced hepatotoxicity in HepG2 cells. Culture cells treated with 25 mM APAP (IC50) and Intracellular fraction of probiotics (IFP) from 3 species: Bifidobacterium longum subsp. longum TISTR 2195, Lactobacillus bulgaricus TISTR 451 or Streptococcus thermophilus TISTR 458. Results showed that IFP of
S. thermophilus TISTR 458 suppressed growth inhibition and cell injury caused by APAP by decreasing lactate dehydrogenase (LDH) enzyme. Additionally, it increased glutathione (GSH) content and decreased enzyme activity of superoxide dismutase (SOD) in HepG2 cells and it exhibited the highest antioxidant activity by ORAC assay. IFP of synbiotics prepared by the incubation of 1% prebiotics (inulin or fructo-oligosaccharide) with probiotics clearly enhanced the potential of IFP in the increasing of enzyme activity in HepG2 cells. Overall, this present study showed that selective probiotics and synbiotics might be helpful in lessening paracetamol-induced hepatotoxicity.