Metabolomic Investigation Of Serum Of Acute Liver Failure Pigs Treated With Fuidized Bed Bioartificial Liver

Part I Dynamic Patterns of Serum Metabolites in Aute Liver Failure PigsBackground&Aims:The study was aimed to explore the Dynamic Patterns of endogenous serum metabolites in the whole process of acute liver failure (ALF), and to develop a metabolomic model for evaluating the severity of ALF.Methods:ALF was induced in pigs (n=8) by intravenous administration of D-galactosamine. Serum samples were collected before the administration,18h,24h and36h after the administration and within12h before death and assayed by ultra performance liquid chromatography-mass spectrometry. The multivariate data was analyzed with SIMCA-P+12.0. Matlab Version7.8(R2009a) was used to perform pattern recognition analysis and construct a linear discriminant analysis model.Results:Three increased patterns and one decreased pattern were found in the whole process of ALF, and each pattern has its own distinct changing trends at different stages. Aromatic amino acids (Patternl) and Lysophosphatidylcholines (LPCs)(Pattern4) might be good markers for evaluating the severity of ALF, while some conjugated bile acids, long chain acylcarnitines (Pattern2) and Glycocholic acid (Pattern3) could indicate liver injury in the early stage. Inspired from the principal component analysis score plot that the pathogenetic condition of ALF aggravated with sampling time, a linear discriminant analysis (LDA) model based on phenylalanine and LPC18:1were further constructed for evaluating the severity of ALF. The leave-one-out cross-validation accuracy of91.67%for the training set and the prediction accuracy of92.31%for the external validation set confirmed its excellent performance.Conclusion:There are at least four Dynamic Patterns in the whole process of ALF, and each pattern has its own distinctive clinical significance. The severity of ALF could be evaluated by the metabolomics model.Part Ⅱ Effects of Fuidized Bed Bioartificial Liver on Serum Metabolites of Acute Liver Failure PigsBackground&Aims:Through characterizing the potential serum metablomics alterations of ALF pigs after the real and sham Fuidized Bed Bioartificial Liver (FBBAL) treatment, this study was aimed to reveal their therapy mechanism and evaluate their safety and get useful information for improving FBBAL.Methods:ALF was induced by intravenous administration of D-galactosamine.18hours later, pigs were treated for6hours as follows. In FBBAL group (B), pigs (n=7) were treated with FBBAL containing encapsulated primary porcine hepatocytes; in sham FBBAL group (S), pigs (n=7) were treated with FBBAL containing cell-free capsules; in ALF group (C), pigs (n=7) were only given intensive care. Serum samples were collected at0h,18h,24h and48h and assayed by ultra performance liquid chromatography-mass spectrometry. The multivariate data was analyzed with SIMCA-P+12.0.Results:Survival time was prolonged significantly (P<0.01) in the B group (70.4±11.5h) as compared with S group (51.6±7.9h) and C group (49.3±6.6h). No apparent metabolomics difference was observed between each group before FBBAL treatment, however, after the treatment, distinct clustering of B group and other two groups, which were intermixed, were presented. Phosphatidylcholines, lysophosphatidylcholines, sphingomyelinase and fatty acids were decreased further, while conjugated bile acids were increased further after FBBAL treatment.Conclusion:Alginate-chitosan capsules have no obvious influence on serum metabolites of ALF pigs. FBBAL possesses some liver functions and could alters the serum metabolome of ALF pigs, and ALF pigs survive longer thereby. Nevertheless, the alterations of metabolites are not universally toward health, and much should be done to improve the therapeutic effect of FBBAL.