| Background:Long-term alcohol abuse can cause a series of liver diseases,including initial simple hepatic steatosis,and subsequent liver inflammation.In general,hepatic steatosis is combined with inflammatory reactions.If effective measures are not taken to relieve liver inflammation,the disease in the liver will further worsen.In addition to gender,age,and ethnic factors affecting the pathogenesis of alcoholic liver disease,the direct toxicity of ethanol and its metabolite acetaldehyde to hepatocytes,and the excessive accumulation of oxygen free radicals during ethanol metabolism affect the structure and function of biological macromolecules.Gut microbial metabolites,and gutderived lipopolysaccharide activate innate immune responses in the liver.Moreover,the innate immune response of liver activated by enteric lipopolysaccharide has attracted more attention.In addition to abstinence,nutritional support,glucocorticoid therapy,there is no good clinical management strategy for ALD.Therefore,it is necessary and meaningful to search for drugs to treat ALD.Leonurus japonicus Houtt,usually named Chinese motherwort,is a plant from the Lamiaceae family.In China,Leonurus japonicus has been used to treat inflammation and gynecological diseases.Leonurine is an alkaloid present in Leonurus japonicus,and previous studies have confirmed that it exerts anti-inflammatory,antioxidant effects and cound relieve abnormal accumulation of fat in nonalcoholic fatty liver in mice.Objective:To explore the anti-inflammatory activity of leonurine on alcoholic steatohepatitis and its partial mechanism through in vitro and in vivo experiments.Methods:In this study,NIAAA model was used to establish alcoholic steatohepatitis model in mice.In simple terms,mice were fed liquid alcoholic diet for 15 days,At the same time,an alcohol gavage was performed on the last day.In terms of grouping,including:alcohol group,control group,positive drug（silymarin）control group,ethanol+LE（40mg/kg）,ethanol+LE（80mg/kg）,ethanol+LE（120mg/kg）,all of which were administered by gavage.In in vitro experiments,RAW264.7 cells were induced by lipopolysaccharide to establish an in vitro inflammation model.Experimental groups:control group,control+leonurine（160μM）group,model（lipopolysaccharide）group,model+leonurine（40μM）group,model+leonurine（80μM）group,model+leonurine（160μM）group.Hematoxylin-eosin staining and oil red staining were used to analyze the lesions of mice liver tissue.The levels of transaminase in the serum of the mice,and the levels of MDA,GSH and SOD in the liver of the mice were measured with the help of biochemical kits,and the ELISA method was used.Changes in the levels of proinflammatory factors in mouse liver were analyzed.Western blotting and qPCR were applied to analyze the changes of related indexes in mouse liver and macrophages.Immunofluorescence and immunohistochemistry were used to analyze the expression of related proteins in cells and liver.level.Analysis of apoptosis and apoptotic protein expression in mouse hepatocytes by Tunel staining and Western blotting.Results:（1）Leonurine can alleviate liver tissue lesions in mice,reduce the content of TG and transaminase in serum,reduce the content of MDA in the liver of mice,and increasethe antioxidant capacity of the liver.（2）ELISA,Western blot,qPCR and Tunel staining showed that Leonurine could reduce the levels of inflammatory factors in alcohol-stimulated alcoholic steatohepatitis and lipopolysaccharide stimulated macrophages.（3）Leonurine cound reduce alcohol induced liver cell apoptosis and improve the antioxidant capacity of liver.（4）Western blot,immunohistochemistry and immunofluorescence results show that leonurine cound reduce the expression of TLR4、P-P65 in liver and macrophages.Conclusion:（1）Leonurine alleviates alcoholic steatohepatitis by reducing apoptosis and oxidative stress of hepatocytes.（2）Leonurine may alleviate alcoholic steatohepatitis through TLR4/NF-κB signaling pathway. |
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