| Objectives:Wilson's disease (WD), which is also called hepatolenticular degeneration(HLD), is a kind of neurological disease due to the metabolism disorder of copper. The objective of the research is as the following:(1)To explore the small mocular metabolites of sub-model, model, TCM treated and penicillamine treated rats of WD, and acquire the related finger-print spectrum and metabolites.(2)To determine and analyze the typical metabolites and possible mechanisms related closely to the sub-model, model, TCM treated and penicillamine treated rats of WD,in order to get subjective data to support TCM theory on this disease.(3) To find a new method for researching WD, which can also serve as a new modernization way for TCM.Methods:66 male Wistar rats (weight=180±20g) were divided into six groups randomly,15-day model group,15-day control group, model group, control group, TCM treated group and penicillamine treated group,with each group containing 11 rats.15-day model group, model group, TCM treated group and penicillamine treated groups were established with excessive copper method. Among them, 15-day model group was administrated with copper food and water for 15 days, while the rest groups for 60 days. Since the 31th day of copper administration, the TCM group was treated with Gandouling tablets by intragastric method, while the penicillamine treated group was treated with penicillamine in the same way. The 15-day control and control groups were naturally taking food and water.All the rats'serum was taken when the above proceedings were over. The serum was tested with 1H-NMR technology. The spectra were edited with MestRe-C2.3 and self-programmed software and then principal component analysis (PCA) was applied to make differentiation between the 15-day model and 15-day control groups, model and control groups,15-day model and model groups, TCM and model groups, penicillamine and model groups, TCM and penicillamine groups respectively. Then, small modular changes in the serum from the above contrast were analyzed to explore the related mechanisms.Results:1,Contrasting the small molecular changes between the 15-day control and 15-day model groups, we found lipids, lactate, proline, and acetone concentrations were lower and glutamine, glucose and phosphorylcholine concentrations were higher in the serum of the 15-day model group.2,Choline and phosphorylcholine concentrations were found to be lower and TAMO+betaine, LDL, VLDL and glucose were significantly lower in the serum of the model group. While creatine+ creatinine and arginine concentrations were found to be higher and lactate, glutamine and glycoprotein were significantly higher in the model group. The small molecular metabolites above may contribute to the discrimination,and serve as references for further research on WD pathogenesis.3,Contrasting the motabolites of 15-day model and model groups, we found the difference lay in the concentrations of lactate, glucose, arginine, acetone and tyrosine.4,Contrasting the small molecular changes between the TCM and model groups, we found glycoprotein,3-hydroxybutyrate,Lipid, glucose,choline,phosphorylcholine concentrations were significantly higher, and leucine, isoleucine concentrations were higher, while lactate concentration was significantly lower, and glutamine Creatine+ creatinine, arginine concentrations were lower in the serum of TCM group.5,Contrasting the small molecular changes between the penicillamine and model groups, we found lactate, glycoprotein, glutamine concentrations were significantly lower, Creatine, creatinine concentrations were lower, glucose concentration was significant higher, and betaine, lipid, choline and phosphorylcholine concentrations were higher in the serum of penicillamine group.6,Contrasting the small molecular changes between the penicillamine and TCM groups, we found 3-hydroxybutyrate, leucine, isoleucine and valine concentrations were higher in the serum of TCM group.Conclusions and highlights:1,It is the first time for NMR based metabonomics to be applied to the research of WD model rats and the comparison of the motabolites after its TCM and western medicine treatment. This research is on the leading of this subject, and is a methodological innovation.2,The small molecular metabolites from the contrast and analysis of the 15-day model and 15-day control,model and control, 15-day model and model groups can be a possible references for the future research on the disease. Among them, the result of glutamine can be possible data reference for nrurological degeneration diseases. Through the analysis of the metabolites, we found the results could coordinate with the TCM interpretation of this disease, which could put forth a new idea for modernization of TCM.3,Through the research on TCM and penicillamine groups, some data could be taken into consideration in the future research on mechanism of penicillamine chelating copper. The detection of some motabolites in the TCM group proved TCM beared the function of nourishing and stenghening liver and kidney.This laid the foundation for researching the curative effect of TCM on WD from a modernized medical perception.4,We explored the difference of TCM and western medicine treating this disease from a systematic facet, through analyzing the difference of the metabolites. As a result, we found the TCM treating WD beared a complex metabolic net and manifested multi-target spot. Which showed metabonomics provided a feasible way for advanced research in the future.5,By experiment, we proved NMR based metabonomics method could be applied to discriminate the sub-model, model rats and control rats, and to discriminate the metabolites after the TCM and penicillamine groups. And through this method, we could get the typical metabolites, which proved this method had a broad application prospect in the future TCM and western medical research of diagnisis, pathophysiology and treatment on WD.
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