GC/MS And LC/MS Based Metabonomic Study Of Liver Diseases

BackgroundThere are lots of reasons can induce liver injury.Hepatitis B is the primary reason in China which can induce multisystemic syndrome.And the incidence of drug induced acute liver damage is increasing with the drug abused.As the most important metabolic organ,the liver takes on lots of functions for the balance of life.Once the liver was damaged,the functions of other organ changed incidentally.Acute liver failure is an intricate syndrome which evolves so quickly after a miserable result to the liver that leads to the development of coagulopathy and hepatic encephalopathy (HE).A great number of middle and small weight molecules involve the procedure of liver failure.However,there are no effective methods to reveal the molecules during the disease happens.The concept about "OMICS" gives the scientists a good idea to study the process of diseases.As the terminal of system biology,metabonomics,defined as "the quantitative measurement of the multiparametric metabolic response of living systems to pathophysicological stimuli or genetic modification",is a novel methodology arising from the post-genomics era for the study of molecular systems biology.So we introduced the metabonomics to study metabolites globally.Methods:The GC/MS technique based metabonomic method was established,and the D-GalN induced liver failure animal serum was introduced to GC/MS.Applying the GC/MS method,the serum samples from hepatitis B induced liver failure patients were collected and detected by the machine.With the principal component analysis,the status of illness was evaluated.Established the approach with Ultra-performance-Liquid Chromatography-ElectroSpray Ionization-quatrupole-Time Of Flight mass spectrometry(UPLC-ESI-Q-TOF)for metabonomic study,and introduced the method to the D-GalN induced liver failure animal serum to judge the state of disease.Detected the metabonomic profiling with UPLC-ESI-Q-TOF in the urine of hepatitis B related liver diseases.Judging the efficacy of the metabonomic data to divide the groups of diseases.ResultsWith the multianalysis to the GC/MS based metabonomic data,the liver failure model could be divided into three groups,one was the control group,one was bioartificial liver support system(BAL)group and the three group is the sham group. 25 metabolites were found by GC-MS.There were amino acids,organic acids and urine et al.Some specific changes in the metabolic composition of serum samples from liver failure patients including amino acids and glucose were shown in GC-MS total ion current(TIC)profiles,and the clear biochemical separation between the healthy persons and liver failure patients was displayed by the pattern recognition method.We also found that the investigated liver failure patients with different grade of severity denoted as model for end-stage of liver diseases(MELD)could be well classified by the corresponding metabonomic data,while the current routine clinical indices,which can also diagnose the liver diseases,are lack of the ability to characterize the global phenotyping of liver failure.With the PLS and CDA,control and hepatitis or hepatocellular carcinoma(HCC)were separated into different groups.Established the UPLC-MS based metabonomic approach.Examination of the metabonomic data obtained by UPLC-ESI-Q-TOF from the individual animals,aided by statistical interpretation of the data,made it possible to group the different individual.The analytical data could be used to define a metabolic "trajectory" as toxicity developed and to provide an explanation for the efficacy by artificial liver support system.With the UPLC-MS,the acquired metabonomic data was analyzed by multivariate analysis.Significant difference in endogenous metabolite profiles was observed in the control and patients with chronc hepatitis B,hepatic cirrhosis and hepatocellular carcinoma.And the abnormality of metabolism recovered the potential biomarkers in diseases.ConclusionsThe results demonstrated that metabonomics is a potential and powerful tool for the characterization of the metabolic perturbation under different physiopathologic status,and also indicated that the metabonomic analysis is promising to provide an integrative criterion to evaluate the severity of the liver diseases and the prognosis.