Metabonomics Studies Of Diseases And Drug Active Mechanisms Based On Chromatography-Mass Spectrometry

Metabonomics is mainly focused on the global metabolic status of biosystems, which reflects the series of biological events that occur under the condition of disease or drug effect by revealing the variation of metabolism. It has been more and more applied to clarify the metabolic characteristic of diseases and the action mechanism of drugs. In this thesis, the metabonomics approach based on chromatography-mass spectrometry was used to study the metabolic disorders of type 2 diabetes and rheumatoid arthritis and the treatment mechanism with berberine for type 2 diabetes and Yishen Juanbi for rheumatoid arthritis.1) First, the metabonomic method on the basis of UPLC-MS and GC-MS was developed, and utilized to analyze the serum metabolic profiles from human participants with normal glucose tolerance (NGT), impaired glucose regulation (IGR) and type 2 diabetes mellitus (T2DM). Multivariate analysis showed the progressive alterations in the metabolic profile from NGT to IGR and further to T2DM with glucose intolerance severity, and IGR was the intermediate status. ANOVA found that many metabolites had significant differences among the three groups. These metabolites variations are related to the dysfunctions in glucose metabolism, lipid metabolism, amino acid metabolism and oxidative stress, which would be closely associated with the development of T2DM.2) A comprehensive metabonomic method in combination with non-targeted and targeted metabolic analysis was performed to reveal the potential mechanism of berberine in the treatment of patients with type 2 diabetes. We found that the concentrations of free fatty acids after berberine administration decreased remarkably and differed from placebo statistically. The results suggest that berberine might play a pivotal role in the treatment of type 2 diabetes through down-regulating the high level of free fatty acids in patient serum so as to improve glucose and lipid metabolism, and reduce insulin resistance.3) A non-targeted metabonomics approach was carried out to profile metabolic characteristics of rheumatoid arthritis (RA) by using LC-MS and GC-MS. The results revealed that the RA patients presented diverse perturbations in inositol phosphate metabolism, lipid metabolism, amino acid metabolism, glucose metabolism, ascorbate metabolism, cholesterol metabolism, glyoxylate and dicarboxylate metabolism. The results are helpful for revealing the complex pathological mechanism in RA. Besides, the ROC curve was utilized to evaluate the accuracy of diagnosis in differential metabolites. The results showed that inositol, glycerate and glycolate had excellent diagnostic performance, and they may become the important clinical indicators of early diagnosis or evaluation drug effects in RA.4) The metabonomic method based on UPLC-MS and GC-MS was performed to study the effects of Yishen Juanbi pill on plasma metabolic profiles of RA model rats in different Chinese syndrome, and explore the anti-rheumatoid arthritis mechanism of Yishen Juanbi. The results showed that Yishen Juanbi pill could significantly improve the metabolic disorders of castration induced-kidney deficiency RA rats, whereas had no repair of pathological metabolism in collagen induced-RA (CIA) rats, hydroxyurea induced-kidney deficiency RA rats and reserpine induced-spleen deficiency RA rats. We found that the mechanism of Yishen Juanbi in the treatment of kidney deficiency in RA involved regulation of abnormal amino acid metabolism, phospholipid metabolism, inositol phosphate metabolism and cholesterol metabolism, and inhibition of inflammatory reaction and oxidative stress.