The Study Of The Hepatoprotective Effect Of The Total Alkaloids From Corydalis Saxicola Bunting Based On Metabonomics And The Metabolism Of Its Active Components

Objectives:1.To reveal the mechanism of the hepatoprotective effect of the total alkaloids of Corydalis Saxicola Bunting based on urinary metabonomics.2.Identify the metabolites of Corydalis Saxicola Bunting in rat plasma,urine,feces and bile after intragastric administration.Methods:1.Forty healthy male SD rats were randomly divided into blank group,model group,TACS group and BP group.Chronic liver injury model was established by subcutaneous injection of CCl₄ peanut oil solution into the model group,TACS group and BP group,once a week for 8 weeks.Rats in the blank group were subcutaneously injected with the same volume of peanut oil solution.The model group and the blank group were given intragastric administration of distilled water.TACS and BP group were given intragastric administration of TACS and BP once a week for 8 weeks respectively.24 h urine samples were collected from rats after the last administration.2.Evaluate the intervention effect of TACS by measuring the activitiesofthealanineaminotransaminase?ALT?,aspartate aminotransaminase?AST?,alkaline phosphatase?ALP?,albumin?ALB?,malondialdehyde?MDA?,superoxide dismutase?SOD?,glutathione?GSH?and histopathologic examination.3.Find the difference of urine metabolites between the model group and the control group,and analyze the possible relationship between the potential biomarkers and the chronic liver injury,based on the metabonomics by nuclear magnetic resonance?NMR?and combined with statistical analysis and online metabolome database.Furthermore,we could characterize the changes of endogenous metabolites in the urine of rats after treatment with TACS,in order to find the potential biomarker and target which may be related to the hepatoprotective effects.4.Using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry?UPLC-Q-TOF/MS?to identify the metabolites of dehydrocarvetin,palmatine and berberine from the CS extract in rat plasma,urine,feces and bile after intragastric administration.5.Using MassLynx 4.1 software and combining with mass depletion filtration?MDF?technique to remove the interference of endogenous substances on the detection of metabolites,so as to analyze the metabolites of dehydrocarvetin,palmatine and berberine in rats.Results:1.The levels of ALT,AST and ALP in serum were decreased significantly?P<0.05?,and the levels of ALB was increased significantly?P<0.05?after treated with TACS;the level of MDA was decreased significantly?P<0.05?,and the level of SOD and GSH were increased significantly?P<0.05?after treated with TACS;the pathological changes of liver in rats were improved after treated with TACS.2.The changes in 2-oxoglutarate,citrate,taurine and hippurate during the process of CCl₄-induced chronic liver injury were significantly restored by TACS treatment,which suggested that TACS synergistically mediated the regulation of multiple metabolic pathways including the TCAcycle,gutmicrobiotametabolismandtaurineand hypotaurinemetabolism.3.It was shown by our results that these compounds was metabolized to the forms of hydroxylation,glucuronidation,sulfation,glucuronidation with sulfation and glucuronidation with hydroxylation in vivo.Conclusions:1.TACS can interventional regulate the disordes of serum biochemicalindexes,livertissuebiochemicalindexesand histopathological changes of rats with chronic liver injury.2.Chronic liver injury induced by CCl₄ could significantly change the metabolic profile of urine,disturb the circulation of TCA cycle,metabolism of intestinal microorganism and metabolism of amino acid.But TACS can regulate the disturbances of metabolic pathways.3.This study could bring valuable insight to evaluating the efficacy of TACS intervention ther-apy,help deepen the understanding of the hepatoprotective mechanisms of TACS and enable optimaldiagnosis of chronic liver injury.4.The metabolites of dehydrocarvetin,palmatine and berberine in plasma,urine,bile and feces of rats are identified.It is of great significance in the identification of absorbed chemical constituents and active component groups,and is an important way to clarify the material basis and mechanism of action of active components of CS...