Metabolomics Study On Qi Stagnationand Blood Stasis Syndrome And Qi Deficiency And Blood Stasis Syndrome Of Coronary Heart Disease Patients With Stable Angina Pectoris

This study was based on a metabolomics analysis method,with stable coronary heart disease and angina pectoris as the research object,and Qi stagnation and blood stasis(QSBS)syndromes and Qi deficiency and blood stasis(QDBS)syndromes as the research syndrome types.Serum and urine samples were collected that met the Western medicine diagnostic criteria and traditional Chinese medicine(TCM)differentiation criteria for coronary heart disease and angina pectoris.Untargeted and targeted analysis methods based on LC-MS/MS technology has established for serum and urine samples of coronary heart disease(CHD)with angina pectoris of different syndromes.This method was used to analyze a large scale of serum and urine samples of CHD with angina pectoris of different syndromes.By comparing the difference between the health people and QSBS syndrome and QDBS syndrome in CHD with angina pectoris,we screened out small molecular markers closely related to the diagnosis of CHD with angina pectoris and analyzed those metabolic pathways.Through the measurement of vascular endothelial function indexes and energy metabolism indexes,the objective indicators and material basis of TCM syndrome differentiation of coronary heart disease are further explored.At the molecular level,the multi-factor regulating mechanism of TCM "syndromes" is clarified,and the essence and scientific connotation of TCM"syndromes" are explored.1.Inclusion and collection of samplesThis chapter focused on the diagnostic criteria,inclusion criteria,and exclusion criteria of the subjects.The gender and age were matched one by one.A total of 90 serum samples and 90 urine samples were collected.There was no statistical difference between gender and age.2.Urine metabonomics of stable coronary heart disease and angina pectoris with two syndrome typesUsing the LC-MS/MS combined technology,non-targeted metabolomics study were conducted on the urine of patients with stable coronary heart disease and angina pectoris.Including urine sample preparation,LC-MS mass spectrometry and optimization of liquid phase conditions,a urine metabolomics analysis method based on LC-MS technology suitable for two syndromes of stable coronary heart disease and angina pectoris was established.In addition,the targeted metabolomics analysis is used for key amino acids.According to the result of PLS-DA,the control group and the patients of coronary heart disease with angina pectoris had a clear separation of the urine metabolic profile,Moreover,there were also significant differences in serum metabolic profiles between patients with QSBS syndrome and patients with QDBS syndrome.Statistical analysis showed that there were 16 differential metabolites.These include abnormalities in the metabolic pathways of amino acids,carnitines,sphingolipids,purines,tricarboxylic acids,and other metabolic pathways.And then clarify the essence of TCM syndromes of coronary heart disease and angina pectoris3.Serum metabolomics of angina pectoris in stable coronary heart disease with two syndrome typesthe combined technology of UPLC and G2-XS QTof mass spectrometer is used to explore the pretreatment of sample,chromatography,and mass spectrometry conditions of untargeted serum metabolomics,and established a stable and reliable LC-MS method for serum samples.Afterwards,the combined technology of UPLC and XEVO-TQS mass spectrometer was used,and the multi-reaction monitoring mode(MRM)method was used to study the serum metabolomics of targeted amino acids and lipids,including the extraction of lipid samples,Selection of shaking time,optimization of reconstitution volume,etc.Samples were applied to the study of combining untargeted and targeted metabolomics.According to the result of PLS-DA,The serum metabolic profile of the control group was significantly different from the patients with coronary heart disease and angina pectoris.Moreover,there were also significant differences in serum metabolic profiles between patients with Qi stagnation and blood stasis syndrome and patients with Qi deficiency and blood stasis syndrome.Statistical analysis revealed that there were 86 differential metabolites.These include abnormalities in metabolic pathways such as amino acids,carnitines,sugars,organic acids,cholic acids,fatty acids,sphingolipids,etc.,and further clarify the essence of TCM syndromes at the molecular level4.Measurement of vascular endothelial function indexes and energy metabolism indexes in serum samplesAll serum samples collected were measured for endothelin-1,6-keto-prostaglandin F1?,soluble intercellular adhesion molecule-1,soluble vascular cell adhesion molecule-1,nitric oxide,ATP and thromboxane B2 in this study.Significant differences were found between the patients with different syndromes of coronary heart disease and healthy controls.The results showed that the content of vasoconstriction factors such as endothelin-1 and thromboxane B2 increased,the content of vasodilators such as nitric oxide and 6-keto-prostaglandin decreased,the content of soluble intercellular adhesion molecule-1 increased,and the content of ATP decreased.These may cause myocardial damage,which can lead to coronary heart disease.