The Study On The Effect And Material Basis Of Alisma Orientale Extract Alleviating Liver Injury In Fructose-induced Mice

Nonalcoholic fatty liver(NAFLD)is a disease closely related to metabolic disorders,include liver fat accumulation and inflammation with insulin resistance as the core.In recent years,with the increase in daily consumption of fructose,the incidence and mortality of NAFLD are increasing year by year in the worldwild.NAFLD begins with simple steatosis and further develops into nonalcoholic steatohepatitis,cirrhosis and liver carcinoma.The formation of nonalcoholic fatty liver induced by fructose may be related to fat accumulation,oxidative stress,and inflammation.There are still difficulties in determining the specific pathological mechanism of NAFLD.Studies have shown that the extract of Alisma orientale can significantly regulate liver lipids,sugar metabolism,protect liver damage,anti-inflammatory,anti-oxidation,anti-fibrosis,etc.,but its pharmacodynamic material basis is not very clear.Therefore,the protective effect and material basis of A.orientale extract against liver injury in fructose-induced mice have been studied in depth.In this experiment,on the basis of fructose-induced liver injury model in mice,after intervention through the administration of A.orientale extract,based on biochemical indicators,liver pathological section and metabolomics research and deep exploration of potential biological information,while obtaining monomer compounds by using chemical separation means and fast characterization of high resolution mass spectrometry,further clarify that the extract of A.orientale can alleviate the liver damage in frucose-induce mice and the substance basis of its efficacy.On the basis of fructose-induced liver injury model in mice,after modeling and giving the different of A.orientale extract(low and high dose)for 13 weeks,respectively.Compared with the model group,there was statistically significant in the degree of decline about the body weight of mice in the high-dose group.The results of the postprandial blood glucose test showed that the A.orientale extract group could significantly reduce the postprandial blood glucose in mice.Compared with the model group,the A.orientale extract can reduce the relative content of serum TC,TG,LDC-C,ALT,UA.The results of liver pathological section showed that the extract of A.orientale could significantly reduce the levels of TC、LPS and TNF-α;at the same time,the A.orientale extract can reduce the relative content of LPS in the small intestine.The result of liver pathological section showed that the fructose water can induce inflammatory cell infiltration,hepatocellular enlargement and steatosis in liver tissue of mice.the A.orientale extract administration group could significantly alleviate the degree of liver inflammation,cell morphology and steatosis.Based on ~1H-NMR liver metabolomics,52 endogenous metabolites were identified from liver tissues,and 32 differential metabolisms(model vs blank)were screened for fructose-induced live injury.The metabolic pathways are mainly enriched in amino acid metabolism,glucose metabolism,lipid metabolism,glutathione metabolism,taurine and hypotaurine metabolism,etc.It is suggested that fructose might induce the disorder of glucose and lipid metabolism,cytokine secretion in the liver of mice.Compared with the model group,the A.orientale extract administration group mainly affected the changes of 26 metabolites,including 21 different metabolites were screened together(model vs blank and the A.orientale extract administration group vs model).Compared with model group,the A.orientale extract administration group can bring 16 metabolites back to normal metabolic levels,which is statistical significance.From the three indexes of serum,tissue biochemical indexes,liver pathological section,and metabolomics,the comprehensive evaluation of the extract of A.orientale can alleviate liver injury in fructose-induced mice.In this experiment,based on phytochemical separation,thin layer chromatography,open chromatographic column,preparation liquid chromatography,high performance liquid chromatography,nuclear magnetic resonance spectroscopy,high resolution mass spectrometry,polarimeter,circular dichroism chromatograph and other methods,10 compounds were isolated and identified from the A.orientale,among them,triterpenoids(alisol C 23-acetate,alisol B23-acetate,alisol F,alisol F 24-acetate,alismaketone 23-acetate),sesquiterpenoids(1S,4R,5R,10S-guaianediol、1S,4S,10S-calamusin I),alkaloid(1H-indole-3-carboxylic acid),fatty acid(hexadecanoic acid)and steroids(β-sitosterol).High-resolution mass spectrometry is used to quickly characterize its chemical composition,36 species were speculated,including33 triterpenoids.Studies have reported that the extract of A.orientale can inhibits simple steatosis,inhibits the secretion of adipokines and cytokines and improves endoplasmic reticulum stress.Among them,alisol B 23-acetate can effectively regulate lipid metabolism,suger metabolism,inhibit oxidative stress,improve liver inflammation and fibrosis.Through the pharmacodynamic evaluation of the extract of A.orientale can alleviating liver injury in fructose-induced mice and the isolation,purification and speculation of its chemical components,it was initially proved that the extract of A.orientale can improve the liver injury in fructose-induced mice.It provides a scientific basis for the prevention,treatment of A.orientale for NAFLD and establishes the foundation for the further research of Chinese medicine A.orientale.