Study On The Bioactive Components In Vivo And The Mechanism Of Treating Hepatochlic Hygropyrexia Syndrome Of Longdan Xiegan Decoction

AIMApplication of UPLC-Q-TOF technology,the chemical components in vitro and the absorbed components in rat plasma after oral administration of Longdan Xiegan Decoction was studied to reveal active substance directly in vivo,which may offer candidate compounds for the study on material base of therapeutic effect.Analysis of the therapeutic effects of LXD on hepatochlic hygropyrexia syndrome model rats based on the urine metabolimic analysis method,the metabolite profiles and the biomarkers of rats of LXD on hepatochlic hygropyrexia syndrome was in depth explored to reveal its mechanism.METHODSAccording to the theory and method of Serum Pharmacochemistry of the traditional Chinese medicine,UPLC-Q-TOF technique was applied to established the chemical components in vitro and the absorbed components in rat plasma after oral administration of Longdan Xiegan Decoction,the chromatograms acquired in vitro and vivo was processed through the intuitionistic comparation to reveal the components absorbed into blood.hepatochlic hygropyrexia syndrome model rats was duplicated by feeding with alcohol mixture combined with high fat and sugar diet with the appearance,bile secretion,physiological and biochemical indexes of histopathological changes.The study evaluated the therapeutical effects and the mechanism in rats of LXD on hepatochlic hygropyrexia syndrome using metabolimic analysis method.RESULTS1.Analysis of the constituents in vitroA systemic analysis of LXD constituents in vitro was established applying UPLC-Q-TOF technology,moreover,111 chemical composition were identified objectively in the majority with flavonoids ingredients,and others iridoids,sapnins,alkaloids and sterols.Among them,12 constituents from Gentiana scabra Bge,11 constituents from Gardenia jasminoides Ellis,5 constituents from Bupleurum chinensis,9 constituents from Alisma plantago-aquatica,25 constituents from Scutellaria haicalensis Georgi,7 constituents from Plantago asiatica L,12 constituents from Radix Angelica Sinensis,19 constituents from Glycyrrhizae,5 constituents from Akebia quinata,3 constituents from Rehmannia glutinosa,otherwise 1 constituents from Scutellaria baicalensis Georgi,Plantago asiatica L.Akebia quinata,Rehmannia glutinosa,1 constituents from Gardenia jasminoides Ellis.Plantago asiatica L,1 constituents from Bupleurum chinensis,Scutellaria baicalensis Georgi.Plantago asiatica L.Akebia quinata2.Analysis of the constituents in vivoThe chromatograms of medicated serum of rats was acquired with optimizing blood collection time of rats and the method of plasma treatment based on analysis method in vitro.Through the intuitional comparison of the retention time and high and low collision energy mass spectrum of dosed rat plasma with those of controlled rat plasma and LXD,25 compounds derived from LXD were detected and characterized in rat plasma,including 21 prototype components and 4 new metabolites3.Effects mechanism research of LXD on hepatochlic hygropyrexia syndrome model ratsWistar rats were treated with alcohol mixture combined with high fat and sugar diet in humid and hot environment to duplicated hepatochlic hygropyrexia syndrome model,the study founded anal temperature rising,the volume of bile secretion decreasing,AST?ALT?ALP?T-Bil?CHOL?TG??-GT?TBA?MDA increasing markedly,SOD lower(P<0.05,P<0.01)after model group rats at 4th week,and liver tissue pathological section appeared fatty liver and fatty degeneration,all statistical results proved hepatochlic hygropyrexia syndrome model was built successfully.The model group rat was treated with LXD after 4th week,the study showed all indexes of the treated rats called back remarkably.A metabolomic approach which was suitable for the rat urine samples was established based on UPLC-Q-TOF optimizing,the urine metabolic profiling and track was analyzed and 40 biomarkers was conformed and identified,then found out these markers related metabolic pathways.After analysis of the data of LXD treatment group,model group and control group,urine metabolic profiling of treatment group were trend to normal,the level of 19 biomarkers in treatment group had obvious trend of callback.These biomarkers which have significant differences were as efficacy biomarkers of LXD on hepatochlic hygropyrexia syndrome model and to reveal its effect mechanism.CONCLUSIONSDepending on UPLC-Q-TOF technology,the study represented 111 LXD constituents in vitro successfully and preliminary identified constituents absorbed into blood in vivo,25 was founded and identified,which provided the basis for further study of LXD effective substances.Hepatochlic hygropyrexia syndrome model was duplicated successfully,all statistical results proved LXD was very efficacy for relieving or eliminating the rat liver inflammation and syntonying various indicators of hepatochlic hygropyrexia syndrome significantly.It demonstrated LXD can be the effective treatment of hepatochlic hygropyrexia syndrome.