Vivo Distribution In Rats And Metabolism Study Of Acteoside

Acteoside is one typical phenylethanoid glycoside with a lot of biological activities.Acteoside extracted from Plantago asiatica L in our laboratory?purity>98%,deteched by HPLC?was found that it was an effective xanthine oxidase inhibitor and a prevention of hyperuricemia,but the research about its metabolism was hysteretic,so this paper focus on tissue distribution in vivo,simulated degradation in vitro,analysis of metabolic product in urine and metabolism pathway based on metabonomics study.The results as follows:1.HPLC was used to detect the content of acteoside in heart,liver,spleen,lung,kidney,stomach,small intestine,large intestine and plasma of SD rats with a single dose of acteoside in 15,30,45,60,90,120 min respectively after the gavage.It was found that acteoside in gastrointestinal was significantly higher than other tissues.Acteoside concentration fell sharply in the digestive tract,caffeic acid was detected and increased at the same time.So it was speculated that acteoside was degraded to caffeic acid in the digestive tract.The intestine metabolite was identified by HPLC-MS for further confirmation and it was confirmed that caffeic acid was the main metabolite of acteoside.In addition,another unknown metabolites was detected,it was speculated the unknown metabolites was metabolism product of acteooside after II phase reaction in intestinal.2.HPLC-MS was used to detect the acteoside metabolic production in simulated gastrointestinal digestive juice in vitro.Results showed that in artificial gastric digestion,hydroxytyrosol,acteoside de-hydroxytyrosol product and isoacteoside three metabolites were detected,degradation rate was 11.62±in 1.78%;in artificial intestinal fluid digestion,caffeic acid and isoacteoside two metabolites were detected,degradation rate was 18.87±1.09%;in artificial gastric and intestinal fluid accumulation digestion,hydroxytyrosol,caffeic acid,de-hydroxytyrosol acteoside and isoacteoside were detected,degradation rate was 26.68±1.38%.3.UPLC-QTOF-MS was used to detect the metabolites in SD rats urine after a single dose of acteoside,and 24 possible acteoside metabolites was found.It was speculated that hydrolysis of glycosidesa was reacted firstly and the corresponding product was hydroxytyrosol?M₁?and de-hydroxytyrosol acteoside(M₁₄).Then ester bond of de-hydroxytyrosol acteoside(M₁₄)was broken and generated to caffeic acid?M₅?;hydroxytyrosol?M₁?and caffeic acid?M₅?was oxidized,reduced,methylated,been sulfuric acid,and been glucose oxidated,obtained a series of other metabolites.4.Based on metabonomics,pre12h?-12h?and 4h,8h,12h and 24h after gavage of acteoside and hyperuricemia model urine of SD rats were analyzed in principal component analysis?PCA?method,combining with website searching,according to the differences and variety of the metabolites to determine the affected metabolic pathway.It was found that normal group,control group and treated group in hyperuricemia model were different,treated group was more similar to normal group,acteoside could treat and prevent hyperuricemia.By monitoring the change of metabolic markers,it was found that the metabolic pathways affected by acteoside on hyperuricemia were:cholesterol biosynthesis,sphingolipids metabolism,glycolysis and gluconeogenesis,lactic acid cycle,nucleotide metabolism,glucose homeostasis,urea cycle and amino compounds metabolism,amino acid and its derivatives metabolism,phase II metabolism,benzoic acid metabolism and amino acid conjugation,sulfated,methylation,one carbon metabolism.When acteoside affecting on normal rats,it was found that the blank control?-12h?and the other four groups was signifiantly different;in addition to the individual group samples,the difference between 4h,8h group and 12h,24h group along the X axis was greater;4h and 8h was different in along the Z axis.12h urine samples and 24 h urine samples were more similar.By monitoring the change of metabolic markers,it was found that the metabolic pathways affected by acteoside on rats were:energy metabolism-TCA cycle,glucose homeostasis,urea cycle metabolism,amino acid metabolism,nucleotide metabolism,nicotine metabolism,phase I metabolism,phase II metabolism and bile acid metabolism;The metabolic pathways were closely related to the function of anti oxidation,anti inflammation and blood uric acid in the body,and it had no toxic and side effects on liver and kidney.