Study Of Phlegm-turbid Syndrome And Blood-stasis Syndrome Of Angina Pectoris Due To Coronary Heart Disease Based On Metabolomics

Background:As the first main cause of death in our country, cardiovascular disease is a complex process closely related to metabolic, with morbidity and mortality increasing. Its occurrence, development associated with metabolite abnormalities in metabolism of body fluids and tissues. Syndrome identification is the key feature of Chinese medicine. The diagnosis, treatment, prescription, medication are begin at syndromes. The gate of medical diagnosis and treatment of coronary heart disease is Chinese Traditional Medicine syndromes. Metabolomics is a new method developed follow with genomics, proteomics to explore the overall metabolic state of the body. The body metabolism disorder caused by disease often show the performance by the metabolic group in body fluids such as urine and blood. Testing and analysis on the metabolism of body fluids such as urine and blood group will contribute: understanding to the biochemical changes accompanying the process of the development of the disease, the disclosure of the early metabolomics markers related diseases and recognization of the molecular mechanism of pathogenesis, and the diagnosis of disease in early stage. Purpose:In this study, metabonomics technology was used for comparison between blood, urine of angina pectoris patients with phlegm-turbid syndrome and blood-stasis syndrome, and among three kinds of blood-stasis syndrome:Qi-stagnation and blood-stasis syndrome (QSBS), heart blood-stasis syndrome (HBS), Qi-deficieney and blood-stasis syndrome (QDBS). We tried to explore syndrome biomarkers underlying patterns of disease by detecting changes of blood and urine metabolites and to provide a scientific basis for angina pectoris phlegm-turbid syndrome and blood-stasis syndrome study. We tried to apply ideas and methods to explore the nature of Chinese Traditional Medicine syndromes, by the establishment of disease and syndrome combining metabolomics research platforms.Methods:1, Plasma metabonomics study for angina pectoris patients with phlegm-turbid syndrome and blood-stasis syndrome:plasma samples from 10 patients with angina pectoris due to coronary heart disease phlegm-turbid syndrome patients and 10 patients with angina pectoris due to coronary heart disease blood-stasis syndrome were detected by nuclear magnetic resonance ('H NMR). Plasma metabolites profilling difference between angina pectoris patients due to coronary heart disease with phlegm-turbid syndrome or with blood-stasis syndrome was analysed by discriminant analysis (PLS-DA) method.2, Urine metabolomics study on angina pectoris patients due to coronary heart disease with phlegm-turbid syndrome, blood-stasis syndrome:urine samples from 12 patients with angina pectoris due to coronary heart disease phlegm-turbid syndrome patients and 12 patients with angina pectoris due to coronary heart disease blood-stasis syndrome were detected by nuclear magnetic resonance ('H NMR). Urine metabolites profilling difference between angina pectoris patients due to coronary heart disease with phlegm-turbid syndrome or with blood-stasis syndrome was analysed by discriminant analysis (PLS-DA) method.3, Plasma metabonomics study for angina pectoris patients due to coronary heart disease with three blood-stasis syndrome:plasma samples from 18 healthy people and 18 patients with angina pectoris due to coronary heart disease with three blood-stasis syndrome (6 cases QSBS,6例HBS,6 cases QDBS) were detected by nuclear magnetic resonance (1H NMR). Plasma metabolites profilling difference between healthy people and angina pectoris patients due to coronary heart disease with three blood-stasis syndrome was analysed by discriminant analysis (PLS-DA) method.Results:The plasma metabolite levels of angina pectoris patients due to coronary heart disease with phlegm-turbid syndrome and blood-stasis syndrome varied significantly. Compared with APPS patients, LDL, VLDL, Lipid,3-hydroxy-butyric acid, Acetone, Lactate, glucose, NAC, aspartate, isoleucine, glutamine and myc-inositol increased in APBS patients, while HDL, unsaturated lipid, Alanine, Citrulline, Arginine, Proline decreased considerably.PLS-DA of urine 1H NMR spectra revealed revealed different metabolic spectra between APPS patients and APBS patients, demonstrated that the metabolic characteristics of the two groups were significantly different. Compared with APPS patients, Citric acid,α-ketoglutarate, cis - aconitic acid, glucose,3 - hydroxybutyric acid, acetone, tyrosine, creatinine, trimethylamine oxide, dimethylamine, hippuric in urine of APBS patients increased clearly, while the bile acid, histidine decreased considerably.PLS-DA of plasma 1H NMR spectra revealed different metabolic spectra between healthy people and APBS patients. Compared with healthy people, citrate, succinate, glucose, glycoprotein, LDL, VLDL,3-hydroxybutyrate, glutamine, proline, leucine increased in APBS patients, while HDL, unsaturated lipid, threonine, histidine decreased considerably. Different metabolic spectra also were revealed between QDBS patients and other APBS patients. Compared with QSBS patients and HBS patients, Citrate,3-hydroxybutyrate, acetone, TMAO increased in QDBS patients, while glucose, NAC, tyrosine, phenylalanine, proline, leucine decreased.Conclusion:Characteristic metabolic products in each groups can be identified by 1H NMR based metabonomics analysis. Metabonomic study is a feasible and promising way to detect the variation of plasma and urine metabonomics of Angina Pectoris patients with different TCM Syndromes. Changes in urinary metabolites can distinguish different Chinese Traditional Medicine syndromes of angina pectoris patients due to coronary heart disease to some extent. The advantage of metabolomics is more convenient to obtain samples, and more easier to test. It can detect small molecule compounds for qualitative and quantitative research, and the up- and down-regulation of these which perhaps in the future can be used in identification of Chinese Traditional Medicine syndromes. Chinese medicine and metabolomics both explore the multi-body factors affecting the overall condition of a pathological change. The relationship between their theory and the results of this study provide a new idea of objective research to TCM Syndrome, and a new method to reveal the nature of TCM Syndrome.1H-NMR pattern of plasma between healthy people and patients with angina pectoris due to coronary heart disease with blood-stasis syndrome can be distinguished, and further determine the difference in metabolite; in three types of angina with blood-stasis syndrome, plasma'H-NMR pattern of QDBS vary greatly to the other groups, which can determine the difference in metabolites. Wherever there is no obvious differences between QSBS and HBS. This provides us a guideline to further explore the nature of Chinese Traditional Medicine syndromes of coronary.