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Aim To investigate the protective effects of gentiopicroside (GPS) on acute alcoholinduced fatty liver.Methods Mice were treated with ethanol (5 g · kg1 of body weight) by gavage every 12 h for a total of three doses to induce acute fatty liver.GPS (40 or 80 mg · kg1) was garaged with ethanol simultaneously for three doses.Administration of GPS significantly prevented the increases of serum ALT and AST caused by ethanol, as well as serum and hepatic TG.Results GPS could significantly prevent ethanolinduced hepatic steatosis and necrosis by H&E and Oil Red O staining.GPS also significantly inhibited lipogenic genes including sterol regulatory element binding transcription factor 1 (SREBP1), fatty acid synthase (FASN) and acetylCoA carboxylase (ACC) in ethanoltreated mice.Additionally, GPS possessed the ability to prevent ethanolinduced acute liver steatosis, possibly through inducing the phosphorylation of AMPactivated protein kinase (AMPK) and liver kinase B1 (LKB1).After treatment with GPS, peroxisome proliferatoractivated receptor α (PPARα) protein expression in mouse liver was recovered as that level of normal mice.Ethanol treatment evoked P2X7r and caspase1 protein expression,while GPS significantly suppressed those protein expressions.GPS may be developed as a potential therapeutic candidate for ethanolinduced hepatic steatosis and inflammation.