Experimental Study In The Assessment On Quality And Function Of Donor Liver By~1H NMR-Based Metabonomics

ObjectiveThe study was intended to establish the university of Wisconsin solution metabolomics experiments based on1H nuclear magnetic resonance (1HNMR) teehnology, to explore new ideas in the assessment of the quality and function of donor liver.Methods20SD rats, according to the protocol of liver graft procurement; the liver grafts were harvested and preserved at0-4℃with the University of Wisconsin solution for0h、4h8h、12h. The1HNMR spectra at different time were collected and analyzed by the unsupervised statistical method of principal components analysis (PCA) and partial least square discriminant analysis (PLS-DA). The analysis results to the two main components (the Principal Component, PC) t[1]/t[2] were plotted (i.e., the score plot, Scores Plot). The results showed the distribution and differences, due to metabolic differences between samples; the corresponding loading plot showed the major metabolites of the cause of these differences. Using the Statistically normalized integral value of the four groups of university of Wisconsin solution samples metabolites, then done the Single-factor analysis of variance (ANOVA) with SPSS17.0, P<0.05was considered statistically significant difference.Results1. Analysised the0,4,8,12h UW solution with PCA, the organ coefficient of rat liver and weight of the No.2(minimum) and No.15(maximum) does not appear Regularity outlier,that can exclude the impact of differences in organ coefficient to the multivariate statistical analysis; 2. Sample metabolite spectral significant changed; the metabolites which saved4h greatest changed. compared with Oh, Raffinose, adenosine, allopurinol, lactobionate was significantly decreased in4h,8h,12h group; succinate, Inosine, the hypoxanthine, lactate, succinate, alanine, glutamic acid, glutamin), choline, ethanol, trimethylamine oxide/betaine (TMA/betaine), phosphorylcholine/glycerol phosphocholine(PC/GPC), glucose was significantly elevated; glutathione changes are less obvious;3. The metabolites were analyzed in20cases of UW solution samples, using1HNMR Combined pattern recognition of PCA and PLS-DA, Oh and4h, Oh and8h, Oh and12h,4h and8h4h and12h, PCA cross-validation score plot and associated factor loadings Figure;8h and12h, and among the three groups (4h,8h,12h) and among the four groups(0h,4h,8h and12h) PLS-DA cross-validation score plot and factor loadings figure showed, UW solution samples at different time points can be better distinguished in the score plot; differences between the two groups of data with the two principal components, each concentrated at each time point, no obvious cross and overlap.Substances caused by the difference between the groups as described below, with Oh, Raffinose, adenosine, allopurinol, lactobionate was significantly decreased in4h,8h,12h group, with the hepatic preservation time prolonged, the amount of these substances is gradually reduced; succinate, Inosine, the hypoxanthine, lactate, succinate, alanine, glutamic acid, glutamin), choline, ethanol, trimethylamine oxide/betaine, phosphorylcholine/glycerol phosphocholine, glucose was significantly elevated.Conclusions1. The metabonomics better reflect metabolic changes in transplant donor liver metabolite with save time extended, providing an important analytical tool to evaluate and judge the quality and function of the transplanted donor liver.2. The use of pattern recognition methods to find potential metabolic markers can be used for transplant donor quality and functional assessment of early diagnosis,of which the lactate, succinate, hypoxanthine, allopurinol, adenosine can be used as potential metabolic markers in the diagnosis of the quality and function of the transplanted graft.The prediction model based on these markers of information architecture can be realized early prediction of transplant donor quality and functionality.