Metabonomic Study On The Patients With Hepatocellular Carcinoma And Functional Dyspepsia

Different physiological and pathological status lead to different biochemical state and metabolomic profiles accordingly. These overall profiles of metabolites reflect hundreds of compounds in many biochemical pathways, and describe the disease characteristics more completely than the other sciences. Physiopathologic stimulation could cause the changes in concentration and rate of endogenous metabolites through various pathways. Therefore, by comparing the metabolic changes of patients, a group of small molecules related with the occurrence and development of disease could be found. As biomarkers, these micromolecular metabolites can be used to assist the diagnosis and classification of disease. Analysis of metabolic pathways is benefit to helping people understand the development process of disease and the pathway of metabolites in patients deeply. Metabonomics is very important for the illustration of the complicated life systems because of the objective and systematic response of the changes in overall metabolites. In recent years, it is widely applied to clinical diagnosis, screening markers, exploration of pathogenesis and drug toxicity, genetic variation, nutrition and other research areas. In this article, nuclear magnetic resonance (NMR) and gas chromatography / time-of-flight mass spectrometry (GC/TOF-MS) were used to probe the metabolic changes in patients with liver disease and functional dyspepsia. This research mainly includes the following two parts:1. NMR-based and GC-MS-based metabonomics were applied to study the metabolic characteristics of patients with hepatitis, cirrhosis and hepatocellular carcinoma (HCC). A group of micromolecular metabolic markers closely related to the occurrence and development of liver cancer then be found and verified. 1H NMR spectra in conjunction with pattern recognition were applied to sudy the collected serum and urine samples. Metabonomics indicated that the serum level of phosphatidylcholine (PtdCho), triglyceride, betaine, and the urine level of creatinine, hippurate was correlated with the development of liver disease. Compare with hepatitis and cirrhosis, the serum level of glutamate, N-acetyl glycoproteins (NAc), lactate, choline, methionine, low-density lipoprotein / very low-density lipoprotein, (LDL/VLDL) and the urine level of taurine, acetate, citrate in the HCC group were changed significantly. GC / TOF-MS and multivariate statistical analysis showed that the serum concentration of phenylalanine in patients increased significantly (P<0.01) and was positively related to disease process. Compared with other groups, there were relatively lower levels of phosphoric acid and tryptophan in HCC serum, and higher levels of butanoic acid and 3-hydroxybutyrate. Different from previous studies, hepatitis, cirrhosis and HCC were concerned simultaneously in this research, and corresponding characteristic spectrums of metabolites were established. Hence, characteristic metabolites were been searched during the development process of liver disease. Metabolites with continuous changes in the process of hepatitis - liver cirrhosis - HCC or characteristic changes in HCC may be potential biomarkers for early diagnosis HCC, and worthy of further study. At the same time, the exploration of molecular mechanisms changing from hepatitis to HCC played an active role in the establishment of early prevention and risk warning systems of HCC. Meanwhile, the analysis of metabolic pathway indicated that citric acid cycle, urea cycle,lipid and amino acid metabolism and other metabolic pathway were abnormal, which provided a data support for the further explanation of the HCC pathogenesis.2. Metabonomics was applied to study the changes in plasma metabolites of female patients with functional dyspepsia (FD). 1H NMR results showed that, compared with the healthy controls, there were relatively higher levels of glycine, 3-hydroxybutyrate, PtdCho, fructose and lower levels of lactate, glutamate, acetate, NAc in FD female patients. Simultaneously, plasma samples were subjected to GC / TOF-MS. As a result, there were about 122 peaks detected, and 31 compounds of which were identified, including sugars, amino acids, fatty acids and organic acids. Multivariate statistical analysis showed that, levels of urea, phenylalanine, cholesterol, stearic acid and citrate were higher, while phosphoric acid, mannitol, oxalic acid and linoleic acid were lower in plasma of FD female patients than in healthy controls. In addition, there were higher level of glycine and lower level of valine in FD patients, which were consistent with NMR results. These results suggested that, changes in plasma metabolites of female patients with FD could be evaluated by metabonomics objectively. Current diagnosis and treatment of FD still be short of objective and reliable evaluation standards. Metabolites with obvious changes in this research have potential to be candidate biomarkers used for the clinic. Analysis of metabolic pathway indicated that several abnormal metabolism happened in vivo of FD female patients, including citric acid cycle, urea cycle,lipid and amino acid metabolism and other metabolic pathway. There are few reports on metabolic characteristics of FD, and most of these reports have paid attention to the effect of acupuncture based on FD patients. Different from previous research, 1H NMR and GC / TOF-MS were applied to observe overall changes of micromolecular metabolites in plasma of FD female patients. Metabonomics provide a unique perspective for the pathogenesis research of FD.