Study On The Differential Expression Profiles And Atherosc Lerosis Molecular Mechanism Of MiRNA-mRNA Genes Related To Phlegm,Fever And Blood Stasis Syndrome Of Coronary Artery Lesion Treated By Huatan-Huoxue-Qingre Decotion

"China Cardiovascular Disease Report 2018" reports that the number and proportion of cardiovascular diseases in China are currently at the forefront,with approximately 290 million patients with coronary heart disease and cardiovascular disease.Mortality rates continue to increase in rural areas,especially acute myocardial infarction(AMI),which has surpassed urban death rates in the past 10 years.The interventional treatment of coronary heart disease has increased by 13% compared with the past,thus causing huge medical expenses.The study found that 30% of patients with coronary stenosis of 40%-70% can be accompanied by myocardial ischemia,and 35% of patients with 50%-70% ischemic degree of FFR?80%.For the history of patients undergoing coronary angiography percutaneous coronary intervention(PCI),7/8 patients had a moderate stenosis range(30%-70%)before the onset of angiography,and 6% within one year Development of acute lesions.This range is currently tentatively defined by the medical community as coronary heart disease.Coronary critical lesion refers to the lesions of coronary stenosis(30%-70%)measured by coronary angiography(CAG)diameter method.It is a special stage in the progression of coronary atherosclerosis and its pathogenesis,Pathological characteristics,modern treatment,TCM diagnosis and treatment are consistent with coronary heart disease(CHD).Studies have shown that the main types of TCM syndromes that are common in coronary lesions are the combined syndromes of phlegm dampness,heat accumulation,and blood stasis.In this study,blood samples from clinical trials of patients with coronary sputum stasis and heat syndrome treated with Huatan-Huoxue-QingreDecotion were collected,and high-throughput sequencing and bioinformatics analysis were performed to find the genetic differences in the combination of disease and syndrome and Huatan-Huoxue-QingreDecotion.Expression spectrum.At the same time,explore the molecular mechanism of Xuanbi Antong Recipe based on the combination of disease syndrome and atherosclerosis.Method 1 Clinical literature researchIn this study,a combination of coronary heart disease,phlegm,stasis and heat syndrome and disease syndrome patterns was developed,and corresponding search languages were developed.The Chinese databases of HowNet,CBM,Weipu,and Wanfang were self-built and published in December 2019.Reporting(clinical trial research and expert clinical experience).The basic characteristics of the literature,the prescriptions of syndromes and treatments,and the canonical traceability are extracted,and in combination with "Treatise on Febrile Diseases and Miscellaneous Diseases",the corresponding prescriptions and corresponding prescriptions are discussed.2.Study on the dosage compatibility of the prescription for removing phlegm,promoting blood circulation and clearing heatOn the basis of the above,combined with the prescriptions and classic articles of Zhang Zhongjing's Treatise on Febrile and Miscellaneous Diseases,the related drugs of Huatan-Huoxue-QingreDecotion were analyzed,and the compatibility rules of the drugs were found.Provide a basis for clinical application of Huatan-Huoxue-QingreDecotion.3 Network pharmacology study on the treatment of syndrome AS with HHQTCMSP,Pubchem,SwissTargetPrediction,and UniProt predict that Huatan-Huoxue-QingreDecotion can effectively regulate targets.TTD,DrugBank,and DisGeNET were used to obtain coronary heart disease targets.GeneCards obtained atherosclerotic(AS)mRNA.Analysis of ClueGO and ClusterONE of Cytoscape 3.2.1,to obtain statistically significant(P <0.01)pathway information of core target participation.Active nodes,predicted intersection targets,action pathways,and pathway annotations are used as nodes,and the Excel tables are used to establish the corresponding relationships,which are imported into Network Analyzer,and the phlegm and blood circulation are constructed according to the degrees of freedom and degree of key topological parameters.Qingre Recipe-"Composition(miRNA)-TargetPathway" Network Diagram for Atherosclerosis of Coronary Heart Disease,through the comprehensive formation of the "Composition-Target-Pathway" network,discovering multi-target,multi-target Approach,multi-level role.The relationship between traditional Chinese medicine ingredients and predicted targets is marked with special colors.By constructing a visual network diagram of "components-targets-signal pathways",it provides a basis for laboratory research.4 Study of differential expression profileA total of 30 exceptions were collected from the treatment group of coronary sputum stasis and heat syndrome(before and after treatment),the coronary sputum stasis and heat syndrome of comfort group(before and after treatment),the non-sputum stasis and coronary stenosis group and the normal group.Peripheral blood nucleated cell samples were tested for the expression of miRNA and mRNA by high-throughput sequencing technology,and comprehensive analysis of non-treatment of coronary sputum stasis-heat syndrome(experimental group,placebo group)and normal group and coronary angiopathy was performed.Comparison of phlegm and stasis fever group,analysis of self-control(post,anterior)of coronary sputum stasis-heat type treatment group and coronary sputum stasis-heat type comfort group(back,anterior)of coronary artery critical lesion.The differentially expressed genes were screened for correlation analysis to obtain miRNA and mRNA differential expression profiles of phlegm and stasis heat syndrome in critical coronary lesions,and miRNA and mRNA differential expression profiles of phlegm and stasis heat syndrome in critical coronary lesions treated with Huatan-Hu oxue-QingreDecotion.5 Study on the molecular mechanism of HHQ in the treatment of syndrome ASTo explore the mechanism of Huatan-Huoxue-QingreDecotion in the treatment of AS-related miRNA and mRNA of coronary sputum stasis-heat syndrome.Obtain the differential gene expression profiles of Huatan-HuoxueQingreDecotion for the treatment of coronary sputum stagnation and heat syndrome,and use GeneCards database to retrieve atherosclerosis-related mRNA and miRNA.Combined with Huatan-Huoxue-QingreDecotion to predict treatment targets,three parts The intersection is considered to be a key gene for the treatment of AS with coronary sputum stasis and heat syndrome by Huatan-Huoxue-QingreDecotion.Cluster analysis,principal component analysis(PCA),GO pathway,and KEGG pathway analysis are used.The miRBase and starBase databases were compared with the annotated sequencing results,and the gene regulatory association was obtained through analysis.A miRNA and mRNA regulatory network for phlegm-stasis-heat syndrome with coronary phlegm stasis and heat syndrome were established through the analysis of Huatan-Huoxue-QingreDecotion.Result: 1 Clinical literature researchAnalyze 49 articles that meet the inclusion / exclusion criteria,16 dissertations,33 journal articles,the earliest year 2000,the latest year 2019,Siwu Tang(Peach Red Siwu Tang)and Xuefu,which are clearly related to Zhongjing Academics Zhuyu Decoction,Wendan Decoction(Huanglian Wendan Decoction),Erchen Decoction,Chaihu Decoction Breast Decoction,Xiaotra Decoction Breast Decoction,Guabai Bai Pinxia Decoction.Decoction,Sini San,Pinellia Poria Decoction,Tangerine Bamboo Ru Soup,Xiao Chai Hu Tang,Xiao Xi Chest Soup,Guabai Bai Pinxia Soup,retrospective analysis.Induction of increased phlegm-stasis-heat syndrome in coronary heart disease is related to obesity,decreased activity,and poor diet and rest.It is a complex disease with blood-stasis syndrome as the core and phlegm-heat syndrome as the main body of deficiency and truth,and is closely related to liver and kidney essence and blood deficiency.Therefore,while promoting blood circulation,clearing heat and resolving phlegm,attention should be paid to correcting deficiency and regulating liver and kidney as appropriate.2 Study on the dosage compatibility of the prescription for removing phlegm,promoting blood circulation and clearing heatAccording to induction,it is found that in the application of Wuwei Chinese medicine decoction,there are 14 prescriptions compatible with Coptis chinensis,44 prescriptions compatible with Pinellia,50 prescriptions compatible with Paeonia,5 prescriptions compatible with Pueraria,and 38 prescriptions with Ginseng.first.There are 32 prescriptions that are compatible with 5 Chinese medicines.In the application of decoction,the usual dose of compatible Huanglian is one or two(57.14%);the commonly used dose of compatible Pinellia is half liter(78.26%),and the usual dose of compatible paeonia It is two two,three two(45.45%),the commonly used dose of compatible pueraria is four two(40%),and the commonly used dose of compatible ginseng is three two(42.11%).In the treatment of coronary heart disease,"Huanglian-Pinellia" enhances heat-clearing,stomach,dryness and dampness,and phlegm,"Ginseng-Pinellia" enhances righting,dispels evil,and reduces inversion,and "Huanglian-Ginseng" purifies the deficiency and nourishes Qi Quxie,"pueraria root-peony" enhances Tongluo and removes stasis and promotes blood circulation.3 Network pharmacology study on the treatment of syndrome AS with HHQ Network analysis shows that Huatan-Huoxue-QingreDecotion Decoction for coronary heart disease AS regulates 82 targets through 129 therapeutic components,and through GEO2 R on-line analysis of multiple gene chips,there are up and down differences in targets,35 targets up and 47 targets down.In 12 biological processes,54 specific pathways.Conclusion: The mechanism of Xuanbi Antong Decoction in treating AS with coronary heart disease may be the regulation of PTGS2?MGLL?PDGFRA?PPARG?NR1H3?PPARA?TNF?SCARB1?IL1B?VDR?NFKB1?LDLR?IL6?APOB through eight Chinese medicines.And lipoprotein synthesis and metabolism,coagulation function,NO biosynthesis process,inflammatory response,lipopolysaccharide response,positive regulation of sequence-specific DNA binding transcription factor activity,negative regulation of external stimulus response,biological homeostasis,etc.The pathway works.4.Study on differential expression profileHigh-throughput sequencing technology and bioinformatics methods have been used to obtain differential expression profiles of coronary sputum stasis and fever syndrome genes,including 557 mRNAs(249 up-regulated and 308 down-regulated)and 162 miRNAs(28 known up-regulated 37 up-regulations were found,38 known down-regulations and 59 new down-regulations were found).High-throughput sequencing technology and bioinformatics methods to obtain differential expression profiles of coronary sputum stasis and fever syndrome genes including 54 mRNAs(38 up-regulated and 16 down-regulated),and miRNAs 33(known 4 up-regulated,new 17 up-regulations were found,5 known down-regulations and 7 new down-regulations were found).5.Study on the molecular mechanism of HHQ in the treatment of syndrome ASThere are 9 AS-related mRNAs and no miRNAs found in the treatment of coronary phlegm phlegm and heat sy ndrome with Huatan-HuoxueQingreDecotion,and 9 differentially expressed miRNAs were obtained through reverse matching,which was further combined with the effective components of Huatan-Huoxue-QingreDecotion prescription to predict the target.Three key mRNAs confirmed by experiments,including PTGS2,MGLL,and PDGFRA,reversely search for miRNAs targeted to target relationships,compare the differential expression of target miRNAs before treatment,and obtain 5 new miRNAs(4 up-regulated and 1 down-regulated),2 known miRNAs(1 downregulation,1 up-regulation),namely novel1020_star upregulation,novel1404_mature upregulation,novel1978_mature downregulation,novel295_mature upregulation,novel926_mature upregulation,hsa-miR-8485 downregulation,hsa-miR-210-5p upregulation Significant statistical difference(P <0.05),found 77 relevant active ingredients.Enrichment analysis of Funrich software,GO pathway,and KEGG pathway revealed that differentially expressed genes participate in 8 biological processes(BP),8 molecular functions(MF),and 12 cellular components,39 specific signal pathways,using Cytoscape3.2.1 software to build molecular mechanism pathway map.ConclusionAt present,there is a type of phlegm stasis and heat syndrome in coronary heart disease,and the main contradiction is the deficiency of truth and evil.The molecular biological characteristics of coronary sputum stasis and fever syndrome were obtained from the differential expression profiles of mRNA and miRNA genes,and the atherosclerosis-related miRNA-The relationship between mRNA regulation and the formation of a "traditional Chinese medicine-component-miRNA-mRNA-pathway-disease" visualization network.The exploratory research of Huatan-Huoxue-QingreDecotion in treating coronary sputum stasis and heat syndrome with “critical component-miRNAmRNA-pathway-disease” provides a basis for future research of this type.