Construction Of Multidimensional Network And Analysis Of Complex Role Of Traditional Chinese Medicine

Research backgroundResearch background:Traditional Chinese medicine(TCM)has been widely used in clinical practice due to its effectiveness,minimal side effects,and suitability for long-term use.However,due to the complex composition of TCM and the complexity of its action hierarchy,it has the disadvantage of unclear mechanism of action,which has resulted in a bottleneck in its clinical application.In this study,we propose a specific biological effect research strategy based on the "characteristic network of disease links" for the research of TCM formulations,including Shengmai Injection,DengzhanShengmai Capsule,Xiaoerfupi Granules,and Sanwujiaowan Pill.It involves the use of experimental data such as proteomics,novel biomarkers,metabolomics,and microbiomics as the basis for construction,bioinformatics tools and literature as the support,and the characteristic component molecular network of TCM formulations as a reference,to construct a multidimensional data network for the complex analysis and clinical optimization of TCM formulations.Research contentsPart 1.Mechanism of Shengmai Injection in treating heart failure with reduced ejection fraction based on "biomarker characteristic network of heart failure with reduced ejection fraction".Using bioinformatics methods,multidimensional data from disease,biomarkers,and drug components were collected to predict the intervention mechanism,molecular targets,and action pathways of Shengmai Injection(SMZ)in treating heart failure with reduced ejection fraction(HFrEF)based on the characteristic network of the disease.The effects and mechanism of SMZ in intervening HFrEF were investigated through in vitro experiments.Based on this,dynamic monitoring of serum indicators in HFrEF rat models was conducted to identify the targets and pathways of SMZ intervention in HFrEF,and several potential biomarkers that can characterize the progression of HFrEF were discovered.Part 2.Efficacy Evaluation and Mechanism Dengzhanshengmai Capsule(DZSMC)in the Treatment of Heart Failure with Preserved Ejection Fraction(HFpEF)based on the"Characteristic Network of HFpEF with Preserved Ejection FractionBased on the research in Part 1 and the specific biological effects of the characteristic components of TCM formulations,a strategy for the three-dimensional complex analysis and clinical optimization of TCM formulations was developed.A disease characteristic network of HFpEF was constructed from multiple dimensions,including drug components,disease targets,and disease pathways,using bioinformatics methods and literature data.The efficacy of DZSMC in treating HFpEF was studied based on this network.Part 3.Analysis of the Complex Effects of Xiaoerfupi(XEFP)on Functional Dyspepsia Based on the "Intestinal Microbiota Disease Characteristic Network"Based on the research foundation of the first two parts and the clear understanding of the pharmacological effects,a "intestinal microbiota disease characteristic network" of functional dyspepsia(FD)was constructed from the levels of brain-gut peptides(BGPs),intestinal microbiota,and metabolites using a rat model of FD,in order to study the multi-layered effects of Xiaoerfupi Granules(XEFP)on FD.Part 4.Clinical Positioning and Complex Action Analysis of Sanwujiaowan Capsule based on the "Clinical Application Characteristic Network":Based on the research foundation of the previous three parts,the "disease characteristic network" strategy was used for clinical optimization and complex action analysis of Sanwu Capsule.A multidimensional characteristic network was constructed based on drug components,target effects,indications,etc.,to optimize the clinical positioning of Sanwu Capsule and study its mechanism for intervention in acute ischemic stroke.Research resultPart 1.Mechanism of Shengmai Injection in treating heart failure with reduced ejection fraction based on "biomarker characteristic network of heart failure with reduced ejection fraction".1.1 Construction of the "Biomarker Characteristic Network of HFrEF":A biomarker-based disease characteristic network containing 603 targets was constructed based on bioinformatics methods,measured proteomics data,and clinical biomarkers.The main pathways included myocardial injury,myocardial pressure overload,myocardial remodeling,renin-angiotensin-aldosterone system disorder,and chronic myocardial inflammation.1.2 Construction of the "SMZ Component-Target" Prediction Network:First,the components of SMZ were determined in TCM-BATMAN,and their potential targets were predicted in SwissTargetPrediction.Heart failure-related targets were collected from Genecard,and bioinformatics analysis was performed to obtain the disease network between SMZ chemical components and heart failure targets.1.3 Prediction of interventions in disease pathways:After bioinformatics analysis of the two networks,the results showed that SMZ played an antiHFrEF role mainly through 17 closely related pathways,which were mainly related to the myocardial fibrosis pathway.Targets included epidermal growth factor receptor(EGFR),apolipoprotein M(APOM),transforming protein RhoA(RHOA),mitogen-activated protein kinase 1(MAPK1),cell division control protein 42 homolog(CDC42),Ras-related C3 botulinum toxin substrate 1(RAC1),and Catenin beta-1(CTNNB1).2.Evaluation of the therapeutic effects and mechanism of SMZ in HFrEF interventionIn this section,the left anterior descending branch of the coronary artery was ligated to establish an acute myocardial infarction model in male Sprague Dawley rats.The therapeutic effect of SMZ on HFrEF was observed,and the results of echocardiography showed that SMZ significantly improved left ventricular ejection fraction and short-axis shortening rate.Masson staining results showed that fibrosis in the left ventricular tissue of heart failure rats was significantly reduced after administration of SMZ.These results indicate that SMZ has a good therapeutic effect on HFrEF and can slow down the degree of myocardial fibrosis.Based on the prediction results of the characteristic network,ELISA was used to detect the levels of EGFR and MAPK1 in the serum.It was found that SMZ could delay myocardial fibrosis by intervening in the levels of these two markers,thus delaying the onset of HFrEF.At the same time,EGFR and MAPK1 could also be used as clinical biomarkers for SMZ intervention in HFrEF.Part 2.Efficacy Evaluation and Mechanism Dengzhanshengmai Capsule(DZSMC)in the Treatment of Heart Failure with Preserved Ejection Fraction(HFpEF)based on the"Characteristic Network of HFpEF with Preserved Ejection Fraction1.Bioinformatics analysis was carried out by searching the Genecard database and combining literature data.It was found that DZSMC mainly works by regulating biological functions such as smooth muscle cell differentiation,regulation of inflammatory response,and response to hormones in the treatment of HFpEF.2.A mouse model of HFpEF was established by mini-pump infusion after nephrectomy and DZSMC was used for intervention.The results showed that after intervention,the hypertrophy of the mouse hearts in the HFpEF model was significantly reduced,and the quality returned to normal levels.HE staining results showed that DZSMC not only improved myocardial hypertrophy but also improved myocardial cell damage and long-term damage caused by excessive myocardial load.The thickness of the left ventricular anterior and posterior walls was calculated from the echocardiography data.After four weeks of continuous drug intervention,the thickening of the left ventricular anterior and posterior walls was improved to varying degrees.This indicates that the abnormal left ventricular function and myocardial hypertrophy caused by long-term disorder of the renin-angiotensinaldosterone system have been alleviated.3.ELISA was used to verify the predicted disease pathways by bioinformatics analysis.It was found that DZSMC can intervene in important pathways of HFpEF,such as slowing chronic systemic inflammation,microvascular inflammation,and reducing myocardial fibrosis.It was found that DZSMC can treat HFpEF by reducing the levels of key molecular targets such as CRP,IL-6,VC AM,E-SELECTIN,and IL1RL1.Part 3.Analysis of the Complex Effects of Xiaoerfupi(XEFP)on Functional Dyspepsia Based on the "Intestinal Microbiota Disease Characteristic Network"1.A rat model of FD was established by administering 0.1%iodine acetamide,and XEFP was used for intervention.The results showed that XEFP significantly improved gastric emptying in the model rats and improved the levels of amylase,lactate dehydrogenase(LD),nitric oxide synthase(NOS),calcitonin gene-related peptide(CGRP),gastrin,motilin(MTL),somatostatin(SS),vasoactive intestinal peptide(VIP),interferon-gamma(IFN-gamma),and interleukin-4(IL-4)in the serum,which were correlated with physiological pharmacological indicators.This indicates that XEFP can treat rat models of FD.2.In the case of a clear understanding of the pharmacological effects,the intestinal microbiota of the rats was sequenced.According to the CAG module analysis,there were significant changes in five categories of microbiota.After adjustment,four categories of microbiota were restored,and the dominant microbiota included Lachnospiraceae,Ruminococcaceae,Streptococcaceae,and Erysipelotrichaceae.Based on bio3.Based on this,non-targeted metabolomics research was conducted on the serum of the rats.The majority of differentially produced metabolites were glycerophospholipids(70.8%),including lysophosphatidylcholines(LysoPC),lysophosphatidylethanolamines(LysoPE),phosphatidylcholines(PC),and phosphatidylethanolamines(PE).Through bioinformatics integration,it was found that XEFP can regulate four microbiota and improve seven metabolic pathways,thereby resetting the balance of metabolites in the rat body and improving FD.Part 4.Clinical Positioning and Complex Action Analysis of Sanwujiaowan Capsule based on the "Clinical Application Characteristic Network":1.Based on the disease characteristic network model,a precise clinical positioning network composed of 324 core proteins for SWJW was constructed according to its clinical indications and databases such as Genecard,BATMAN-TCM,and SwissTargetPrediction.Bioinformatics analysis showed that SWJW scored high in the treatment of stroke and had potential for further development.2.Based on this,a rat model of stroke was established by middle cerebral artery occlusion surgery.It was found that SWJW can reduce the infarct area of the model rat’s brain and improve its behavioral score,and has a certain therapeutic effect on stroke rats.Serum levels of SOD,MDA,AchE,and Ach in the rats were tested according to the predicted results,and it was found that SWJW can regulate these indicators to exert therapeutic effects.3.Proteomics and serum metabolomics analyses were performed on rat brain tissue.It was found that SWJW can improve neuroinflammation,oxidative stress and other biological processes by improving Tau-related proteins.Western Blot technology was then combined,and it was found that SWJW can regulate the levels of phosphorylated Tau protein and Camk2 to improve stroke.It was also discovered that phosphorylated Tau protein(p-Tau)during the acute phase of stroke has potential neuroprotective effects.Conclusion1.Prove that Shengmai injection can delay the occurrence of HFrEF by inhibiting the progression of myocardial fibrosis.2.It is proved that DZSMC can improve the cardiac function of HFpEF mice by adjusting E/A,E/e’ values,alleviating left ventricular hypertrophy,improving the synchronization of heart beat,improving the inflammatory response and microvascular inflammation of mice.3.Combined with the verification results of metabolomics,it is found that XEFP can regulate 7 metabolic pathways to improve the level and balance of 20 metabolites by intervening in four key intestinal flora(Ruminococcae,Streptococcaceae,Erysipelotrichaceae,Lachnospiraceae)and improve a series of BGPs and inflammatory markers to treat FD.4.SWJW can regulate the changes of metabolites such as uric acid,kynurenic acid,3hydroxyphenyl acetic acid,citrate and their related proteins,as well as the changes of Camk2 and pTau,so as to play an anti-stroke effect.