| QI deficiency and blood stasis syndrome is an important factor in ischemic cardiovascular and cerebrovascular diseases.At present,some progress has been made in the research on the biological basis of blood stasis syndrome.However,the research on the occurrence and evolution mechanism of blood stasis syndrome is not yet fully clear,and the biological mechanism of QI deficiency and blood stasis syndrome in ischemic cardiovascular and cerebrovascular diseases still needs further research.Furthermore,compared to body fluids such as serum and plasma,the mechanism closely related to the syndrome of QI deficiency and blood stasis is more likely to be reflected in platelets.Therefore,this study uses metabonomics to explain the mechanism of the syndrome of QI deficiency and blood stasis related to platelets in ischemic cardiovascular and cerebrovascular diseases.ObjectiveA clinical sample study is conducted using metabolomics methods on the Qi deficiency and blood stasis syndrome of ischemic cardiovascular and cerebrovascular disease.The biological basis of the Qi deficiency and blood stasis syndrome of coronary heart disease and the Qi deficiency and blood stasis syndrome of cerebral infarction is characterized as a whole。And a disease syndrome combination animal model similar to clinical practice is established to conduct quasi-clinical research and compare with clinical research results。We need to pay attention to the correlation between clinical research and basic research,and explore the same metabolic pathways and differential metabolites of the two diseases under the same syndrome.At the same time,adopt the research method of "measuring syndrome by formula,corresponding to syndrome by formula",and use Buyang Huanwu Tang to intervene,explore its mechanism of action,and finally explain the relevant mechanism of Qi deficiency and blood stasis syndrome in ischemic cardiovascular and cerebrovascular disease from the perspective of platelets,as well as the mechanism of action of Buyang Huanwu Tang.MethodsFrom the perspective of combining disease and syndrome research,this study adopts metabolomics methods to analyze the platelets and plasma of clinical patients with coronary heart disease Qi deficiency and blood stasis syndrome and cerebral infarction Qi deficiency and blood stasis syndrome.The research focuses on platelets and focuses on key metabolic pathways in platelets.Further at the level of basic research,construct an animal model combining disease and syndrome to preliminarily validate the clinical research results,and explain the intervention effect of Buyang Huanwu Tang as auxiliary validation.By combining clinical and basic research,as well as integrating the same syndrome with different diseases,we can ultimately explore the relevant mechanisms of platelets in the Qi deficiency and blood stasis syndrome of ischemic cardiovascular and cerebrovascular disease.The topic is divided into five parts:The first and second parts are clinical and basic research on the syndrome of Qi deficiency and blood stasis in coronary heart disease.Using metabolomics techniques,platelet and plasma samples from clinical patients with coronary heart disease Qi deficiency and blood stasis syndrome and rat models with coronary heart disease Qi deficiency and blood stasis syndrome are studied to explore the differential metabolites and metabolic pathways affected in the platelets and plasma of patients with coronary heart disease Qi deficiency and blood stasis syndrome and animal models,and to screen for commonly related differential metabolic pathways in clinical and animal models,Explore the pathogenesis of related diseases and syndromes from both clinical and fundamental perspectives.The third and fourth parts are clinical and basic research on Qi deficiency and blood stasis syndrome of cerebral infarction.The research method is the same as the first and second parts.The fifth part is the study of platelet lipid omics in clinical patients with coronary heart disease with Qi deficiency and blood stasis syndrome and patients with cerebral infarction with Qi deficiency and blood stasis syndrome.Based on previous metabolomics studies,focusing on significant changes in lipid related pathways in platelets,quantitative lipidomics analysis was performed on clinical platelet samples using a lipidomics method.Explain the abnormal metabolism of lipid components and the changes in platelet lipid composition ratio caused by two diseases with the same syndrome but different diseases in clinical practice,and reveal the relevant mechanisms of qi deficiency and blood stasis syndrome in ischemic cardiovascular and cerebrovascular disease from the perspective of platelet lipids.ResultsPart 1:There are 13 differential metabolic pathways in platelets of patients with coronary heart disease with qi deficiency and blood stasis syndrome,of which there are 5 main differential metabolic pathways（P<0.05 and impact>1）:linoleic acid metabolism,arachidonic acid metabolism,α-Linolenic acid metabolism,glycerol phospholipid metabolism,and sphingolipids metabolism.Except for arachidonic acid metabolic pathway,other pathways of these five main differential metabolic pathways in platelets are consistent with abnormal metabolic pathways in plasma,indicating that there is a strong correlation between blood and abnormal metabolic pathways in platelets in patients with qi deficiency and blood stasis syndrome.Comparing the biomarkers on the metabolic pathways of plasma and platelets,the common components are mostly lipids,indicating a strong correlation between the abnormal metabolic pathways of blood and platelets in patients with Qi deficiency and blood stasis syndrome.The main abnormal metabolic pathways with significant differences in clinical platelet metabolomics results are all lipid components,indicating significant metabolic changes in lipid components in platelets.Part 2:Through sleep deprivation and coronary ligation of the heart,an animal model of coronary heart disease with qi deficiency and blood stasis syndrome was established.The successful establishment of the model of coronary heart disease with qi deficiency and blood stasis syndrome and the Buyang Huanwu Decoction can effectively intervene in coronary heart disease with qi deficiency and blood stasis syndrome were confirmed from the three aspects of TCM syndrome indicators,disease indicators and platelet parameters.The metabolomics results show that there are 23 differential metabolic pathways in the platelets of rats with coronary heart disease Qi deficiency and blood stasis syndrome,of which 12 are the main differential metabolic pathways:linoleic acid metabolism,sphingolipids metabolism,purine metabolism,glycerol phospholipid metabolism,TCA cycle,glutamine and glutamic acid metabolism,arginine biosynthesis,pentose and glucuronic acid ester mutual conversion,glyoxylic acid and dicarboxylic acid metabolism,alanine,aspartic acid and glutamic acid metabolism,The pentose phosphate pathway and the biosynthesis of phenylalanine,tyrosine,and tryptophan.The metabolic pathways of platelets and plasma overlap more,but multiple amino acid and purine metabolic pathways are enriched in platelets.Comparing the biomarkers of different metabolic pathways between platelets and plasma,the same endogenous components are mostly fatty acids,amino acids,glycerol phospholipids,and sphingolipids,indicating a certain correlation between plasma and abnormal platelet metabolism in coronary heart disease with Qi deficiency and blood stasis syndrome.The first and second parts are clinical and basic research on the syndrome of Qi deficiency and blood stasis in coronary heart disease.The use of metabolomics techniques was used to study platelet and plasma samples from patients with coronary heart disease Qi deficiency and blood stasis syndrome and rats with coronary heart disease Qi deficiency and blood stasis syndrome.Discover 13 differential metabolic pathways in platelets of patients with coronary heart disease with qi deficiency and blood stasis syndrome,including linoleic acid metabolism,arachidonic acid metabolism,α-Linolenic acid metabolism,glycerol phospholipid metabolism,sphingolipid metabolism,with the exception of arachidonic acid metabolism pathway,other pathways are consistent with abnormal plasma metabolism pathway.Comparing the biomarkers on the metabolic pathways of plasma and platelets,the common components are mostly lipids,indicating a strong correlation between the abnormal metabolic pathways of blood and platelets in patients with Qi deficiency and blood stasis syndrome.The key abnormal metabolic pathways with significant differences in clinical platelet metabolomics results are all lipid components,indicating significant metabolic changes in lipid components in platelets.There are 23 differential metabolic pathways in the platelets of rats with coronary heart disease Qi deficiency and blood stasis syndrome.The main differential metabolic pathways in platelets are the biosynthesis of phenylalanine,tyrosine,and tryptophan,linoleic acid metabolism,glutamine and glutamate metabolism,sphingolipids metabolism,glycerol phospholipid metabolism,alanine,aspartate,and glutamate metabolism,TCA cycle,arginine biosynthesis,purine metabolism,conversion of pentose and glucuronic acid esters,glyoxylate and dicarboxylic acid metabolism,and pentose phosphate pathway.The metabolic pathways of platelets and plasma overlap more,but they are more enriched in multiple amino acid metabolic pathways and purine metabolism.Comparing the biomarkers of different metabolic pathways between platelets and plasma,the same biomarker components are mostly fatty acids,amino acids,glycerol phospholipids,and sphingolipids.Among different biomarkers,amino acids and purines are more abundant in platelets,indicating a certain correlation between plasma and abnormal platelet metabolism in coronary heart disease with qi deficiency and blood stasis syndrome.However,there are also some differences,and amino acid metabolism is more prominent in this platelet.Part 3:There are 8 differential metabolic pathways in platelets of patients with Qi deficiency and blood stasis syndrome in clinical cerebral infarction,among which 4 are the main differential metabolic pathways,namely linoleic acid metabolism αLinolenic acid metabolism,glycerol phospholipid metabolism,ether lipid metabolism.Compared with plasma metabolic pathways,the key metabolic pathways shared by plasma and platelets are linoleic acid metabolism,glycerol phospholipid metabolismα-Linolenic acid metabolism.Compared with the biomarkers on the metabolic pathways of plasma and platelets,the common components are mostly lipids,with significant lipid metabolism disorders,which play an important role in the syndrome of cerebral infarction with qi deficiency and blood stasis.Part 4:Through sleep deprivation and multiple cerebral infarction surgery,the animal model of the combination of disease and syndrome of cerebral infarction with stagnation of qi and blood stasis syndrome was constructed,and the intervention effect of Buyang Huanwu Decoction was confirmed from TCM syndrome,disease and platelet indicators.Metabolomics analysis found that there are 23 differential metabolic pathways in the platelets of rats with cerebral infarction Qi deficiency and blood stasis syndrome,among which 8 are the main differential metabolic pathways,including linoleic acid metabolism,sphingolipids metabolism,glycerol phospholipid metabolism,arginine biosynthesis,purine metabolism,taurine and subtrocrine metabolism,glutamine and glutamate metabolism,cysteine and methionine metabolism.The main differential metabolic pathways shared by plasma and platelets are linoleic acid metabolism,sphingolipids metabolism,glutamine and glutamate metabolism,and arginine biosynthesis.Comparing the biomarkers of different metabolic pathways between platelets and plasma,it was found that there are more fatty acids,glycerol phospholipids,sphingolipids,and amino acid components in plasma and platelets,indicating a certain correlation between plasma and abnormal platelet metabolism in coronary heart disease with Qi deficiency and blood stasis syndrome.The third and fourth parts are clinical and basic research on cerebral infarction with Qi deficiency and blood stasis syndrome.The data from these two parts were integrated and analyzed,and the results showed that the quasi clinical differential metabolic pathways of the animal model of cerebral infarction with Qi deficiency and blood stasis syndrome include linoleic acid metabolism and glycerol phospholipid metabolism,α-Linolenic acid metabolism,sphingolipid metabolism,taurine and subtropine metabolism,arachidonic acid metabolism,and unsaturated fat acid biosynthesis.The research results are basically consistent with the clinical differential metabolic pathways of the animal model of coronary heart disease with qi deficiency and blood stasis syndrome.Part 5:Exploring and summarizing the pathogenesis of Qi deficiency and blood stasis syndrome through platelet targeted lipid methods.Among them,there are 55 lipid components that are jointly regulated by the Qi deficiency and blood stasis syndrome of coronary heart disease and the Qi deficiency and blood stasis syndrome of cerebral infarction,mainly involving abnormalities in several lipid metabolism pathways such as PC,PE,TG,PG,SPH,SM,Cer,etc.The pathogenesis of Qi deficiency and blood stasis syndrome may be related to this and directly reflect abnormalities in platelet lipid composition in cardiovascular and cerebrovascular diseases.Among them,the most obvious metabolic changes are in sphingolipids,which involve different components such as SPH,SM,Cer in the metabolic pathway.For the first time,abnormal metabolism of these components and the changes in platelet lipid composition caused by them may be the key mechanism for the pathogenesis of heart and brain qi deficiency and blood stasis syndrome.Conclusion1.The metabolomics results of clinical cardiovascular and cerebrovascular diseases with Qi deficiency and blood stasis syndrome show that there are a total of 8 differential metabolic pathways in platelet samples of cerebral infarction with Qi deficiency and blood stasis syndrome,while there are a total of 13 differential metabolic pathways in coronary heart disease with Qi deficiency and blood stasis syndrome.Among them,a total of 7 identical metabolic pathways show a certain degree of similarity in the impact of the same syndrome and different diseases on platelets,including linoleic acid metabolism α-Linolenic acid metabolism,arachidonic acid metabolism,glycerol phospholipid metabolism,biosynthesis of unsaturated fat,sphingolipid metabolism,taurine and sub taurine metabolism;The results showed that the lipid group related metabolism in platelets with clinical Qi deficiency and blood stasis syndrome was most significantly affected.The lipidomics results of clinical cardiovascular and cerebrovascular diseases with Qi deficiency and blood stasis syndrome show that two diseases with the same syndrome have a strong correlation.The results show that the two diseases are jointly regulated by 55 lipid components,mainly involving abnormalities in several lipid metabolic pathways such as PC,PE,TG,PG,SPH,SM,Cer,and directly reflecting abnormalities in platelet lipid composition in cardiovascular and cerebrovascular diseases.Among them,the most obvious changes in sphingolipids metabolism involve different components such as SPH,SM,Cer in the metabolic pathway.The pathogenesis of Qi deficiency and blood stasis syndrome may be related to this.Components that reflect changes in lipid composition,such as SPH（d17:0）,SPH（d19:0）,SM（d40:1）,SM（d42:2）,SM（d34:1）,SPH（t17:0）,SM（d34:1）,TG（18:310:2-11:4）,etc.2.The metabolomics results of ischemic cardiovascular and cerebrovascular disease with Qi deficiency and blood stasis syndrome in animal models of disease and syndrome combination show that more metabolic pathways are affected in platelets than in clinical samples,and lipid related metabolic pathways are more closely related to clinical relevance.The main differences in platelet samples between animal models of Qi deficiency and blood stasis syndrome of cerebral infarction and coronary heart disease are 12 metabolic pathways in the coronary heart disease group and 8 pathways in the cerebral infarction group.The results showed that the abnormal metabolism of linoleic acid,sphingolipids,and glycerol phospholipids related to lipid metabolism in platelets may be related to the occurrence of blood stasis;The metabolism of taurine and taurine,TCA cycle,pentose phosphate pathway,glutamine and glutamate metabolism can all be involved in energy metabolism,which may be a manifestation of abnormal energy metabolism in platelets of animal models combined with disease and syndrome;Arginine biosynthesis is associated with cardiovascular and cerebrovascular diseases;Purine metabolism is associated with abnormalities in uric acid and oxidative stress.The metabolomics results systematically reflect the changes in platelets of Qi deficiency and blood stasis syndrome.3.Using the research method of "measuring the syndrome by formula and corresponding to the syndrome",the application of Buyang Huanwu Decoction for intervention proves that Buyang Huanwu Decoction can effectively treat cardiovascular and cerebrovascular diseases of qi deficiency and blood stasis syndrome.The metabolic pathways of platelet intervention are glycerol phospholipid metabolism,sphingolipid metabolism,linoleic acid metabolism,unsaturated fat acid biosynthesis,arachidonic acid metabolism,purine metabolism,arginine biosynthesis,glutamine and glutamic acid metabolism,metabolism of alanine、aspartic acid and glutamate. |
PDF Full Text Request