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

ObjectiveThe study was intended to establish the cold preservation model of liver grafts based on 1H nuclear magnetic resonance (1HNMR) technology in clinical experiment, which purpose was to analysis metabolic alternations of liver grafts in different stage with cold preservation to look for new ideas in the assessment of the quality and function of liver grafts.Methods1、40 liver grafts within old preservation was randomly selected. The age of donor was controlled in less than 50 years old, which gander of donor was randomly assigned.Effluent and perfusate samples were collected after 0,4,8,12h of reperfusion. They were divided into 4 groups (0h、4h、8h、12h). Each group has 10 samples. Livers were cold flushed at 4℃ in situ via the portal vein with the preservation solution and preserved at Liquid nitrogen. Perforate samples were studied by H-NMR spectroscopy. In the H NMR study, spectra were acquired at 600 MHz on a Varian UNITY INOVA spectrometer. The Fourier transform was performed on 15000 data points without filtering. Spector was composed of free induction decay signal, and the data is used for pattern recognition analysis, such as unsupervised principal component analysis (PCA), partial east-squares-discriminant analysis (PLS-DA) and orthogonal partial least-squares-discriminant analysis (OPLS-DA). The forms of the results were scores plot and loadings plot. Results were expressed as means± standard error of the mean (SEM) and compared by the use of variance analysis. Significant results were defined by p<0,2、The study was intended to establish the cold preservation model of liver grafts based on 1H nuclear magnetic resonance (1HNMR) technology in clinical experiment, which purpose was to analysis metabolic alternations of liver grafts in different stage and identify resonances.Results1、Sample metabolite spectral significant changed.Raffinose, adenosine, allopurinol, Citrate, Succinate, inosine, lactate, and creatine was significantly decreased in 4h,8h,12h group; glutathione changes are less obvious.2、The data was used for pattern recognition analysis, such as unsupervised principal component analysis (PCA), partial east-squares-discriminant analysis (PLS-DA) and orthogonal partial least-squares-discriminant analysis (OPLS-DA).10 samples were obviously gathered at Oh, and the rest of the sample was dispersed. With the extension of time, the metabolic changes were more obvious. Between the 4h,8h, andl2h spectroscopy samples has some overlap.3、The data of between the 4h,8h, and 12h spectroscopy samples was used for pattern recognition analysis, such as unsupervised principal component analysis (PCA), partial east-squares-discriminant analysis (PLS-DA) and orthogonal partial least-squares-discriminant analysis (OPLS-DA). The main resonances identified were those arising from allopurinol, citrate, succinate, Inosine, Lactate, and creatine.Conclusions1、The study was intended to establish the cold preservation model of liver grafts based on 1H nuclear magnetic resonance (1HNMR) technology in clinical experiment, which purpose was to analysis metabolic alternations of liver grafts in different stage and identify resonances. The main resonances identified were those arising from allopurinol, citrate, succinate, Inosine, Lactate, and creatine.2、There is a close relationship between the main resonances and tricarboxylic acid cycle, Anaerobic glycolysis, Amino acid metabolism, and Purine metabolism.