Analysis method for treating ejection fraction retention type heart failure by using Qihong capsule based on network pharmacology

Abstract

The invention discloses an analysis method for treating ejection fraction retention type heart failure by using a Qihong capsule based on network pharmacology. The method comprises the following steps: step 1, screening active ingredients and core targets of the Qihong capsule; 2, acquiring ejection fraction retention type heart failure gene targets; step 3, constructing a Qihong capsule-HFpEF protein-protein interaction network; 4, gene ontology function enrichment analysis and Peking Doudu gene and genome pathway enrichment analysis, through a method of combining network pharmacology and clinical verification, the curative effect and the action mechanism of the Qihong capsule on HFpEF are discussed, and the result shows that the Qihong capsule can regulate cGMP / PKG signal pathways and various inflammation-related pathways, and has a good application prospect. The traditional Chinese medicine composition shows good treatment potential in the aspect of improving the heart function of an HFpEF patient, provides a scientific basis for clinical popularization and modern application of traditional Chinese medicines, provides more objective basis for treatment of HFpEF by a Qianhong formula, and lays a foundation for deep research on treatment of HFpEF by a qi-tonifying and blood-activating method.

Description

TECHNICAL FIELD

[0001] The application relates to the technical field of biological medicine, and particularly relates to an analysis method for treating heart failure with preserved ejection fraction (HFpEF) by Qihong capsules based on network pharmacology. BACKGROUND

[0002] Chronic heart failure is the end-stage of various cardiovascular diseases, and has a high morbidity and mortality [1]. Heart failure with preserved ejection fraction (HFpEF) is a type of heart failure characterized by normal left ventricular ejection fraction but impaired diastolic function, and the quality of life of patients is significantly reduced and the treatment options are limited. The main manifestations of HFpEF are the presence of symptoms or signs of heart failure, left ventricular ejection fraction greater than or equal to 50%, and B-type natriuretic peptide (BNP) greater than 35 ng / L or N-terminal pro-B-type natriuretic peptide (NT-proBNP) greater than 125 ng / L [2]. Due to the heterogeneity and complexity of its pathogenesis and comorbidities, there is currently no effective treatment for HFpEF in clinical practice. Vascular endothelial inflammation activation is closely related to diastolic left ventricular dysfunction and may lead to the occurrence of HFpEF [3]. The cyclic guanosine monophosphate (cGMP) / protein kinase G (PKG) signaling pathway plays a key role in improving the left ventricular diastolic function of HFpEF patients by enhancing the function of vascular endothelium, reducing the stiffness of myocardial cells and improving calcium ion metabolism [4]. Therefore, the cGMP / PKG pathway may become a new target for the treatment of HFpEF.

[0003] Traditional Chinese medicine has shown potential value in the treatment of HFpEF, and can exert good efficacy through multiple pathways, multiple targets and multiple mechanisms. Qihong capsules, a representative formula of tonifying qi and activating blood, are a hospital agreement formula of traditional Chinese medicine developed by Professor Wang Xiaofeng, a national traditional Chinese medicine inheritance instructor, based on the academic thought of “treating phlegm and blood stasis together” of national medical master Shen Baofan and combined with clinical experience. Qihong capsules have been used for the treatment of heart failure for more than 20 years and have achieved remarkable effects [5]. The previous studies of the research group have explored the mechanism of Qihong capsules in chronic heart failure from the aspects of regulating energy metabolism, anti-inflammation and improving myocardial oxygen consumption [6, 7], but the mechanism of Qihong capsules in HFpEF has not been involved.

[0004] The application designs an analysis method for treating heart failure with preserved ejection fraction (HFpEF) by Qihong capsules based on network pharmacology. By using the network pharmacology method [8], the key targets and signaling pathways of Qihong capsules in the treatment of HFpEF are preliminarily predicted, and the clinical efficacy and potential mechanism are further verified based on the results, so as to provide more objective basis for the treatment of HFpEF by Qihong capsules and lay a foundation for the further research on the treatment of HFpEF by the method of tonifying qi and activating blood. SUMMARY

[0005] The purpose of the present application is to provide a network pharmacology-based analysis method for treating heart failure with preserved ejection fraction (HFpEF) with Qihong capsules to solve the problems raised in the background art.

[0006] The present application explores the potential mechanism of Qihong capsules in treating heart failure with preserved ejection fraction (HFpEF) through network pharmacology, and verifies its efficacy and mechanism through clinical trials. The network pharmacology analysis method is used to obtain the action targets of Qihong capsules and the related genes of HFpEF, construct a protein-protein interaction (PPI) network and obtain the core targets. The effective action targets of Qihong capsules in treating HFpEF are obtained through gene ontology (GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. The heart function and diastolic function of HFpEF patients are evaluated by observing the effects of Qihong capsules on NT-proBNP, E / A value, and E / e' value. The expression levels of serum inflammatory factors and cyclic guanosine monophosphate (cGMP) are detected to explore the mechanism of action. The present application obtains 82 effective chemical components of Qihong capsules, 877 effective targets, 415 HFpEF-related targets, and 108 common targets, among which there are 15 core targets. The GO analysis results show that Qihong capsules mainly regulate biological processes such as vasodilation, inflammatory response, and positive regulation of the MAPK cascade. The KEGG pathway enrichment mainly involves the cGMP-PKG signaling pathway, AGE-RAGE signaling pathway, and lipid and atherosclerosis pathway. In the clinical trial, it was found that the NT-proBNP level of patients in the Qihong capsule group was significantly reduced (P<0.05), the E / A value and E / e' value showed an improvement trend (P>0.05), the serum IL-1β and IL-6 levels decreased (P<0.05), and the cGMP level increased (P<0.05), all of which were more significant than the control group. Conclusion: Qihong capsules can improve the heart function of HFpEF patients, and its mechanism may be related to the regulation of the cGMP / PKG signaling pathway and the improvement of vascular inflammation.

[0007] To achieve the above-mentioned purpose, the present application provides the following technical solutions:

[0008] A network pharmacology-based analysis method for treating heart failure with preserved ejection fraction (HFpEF) with Qihong capsules, the method steps being as follows:

[0009] Step one: effective components of Qihong capsule and core target screening;

[0010] Step two: obtaining of heart failure with preserved ejection fraction gene target;

[0011] Step three: Qihong capsule-HFpEF protein-protein interaction network construction;

[0012] Step four: gene ontology function enrichment analysis and kyoto gene and genome encyclopedia pathway enrichment analysis.

[0013] As a further scheme of the application: the specific steps of step one are as follows: first, collect and sort the compounds of Astragalus, Ramulus Cinnamomi, Rhodiolae, Salviae Miltiorrhizae, Alismatis, Semen Lepidii from TCMSP database, China National Knowledge Network, Wanfang Data, PubMed literature, further query the ADME parameters of these compounds through TCMSP and admetSAR databases, screen effective compound components with oral bioavailability (OB) ≥ 30% and drug-likeness (DL) ≥ 0.18, then find the SMILES structure of these effective compounds through PubChem website, and import them into SWISS TargetPrediction database to obtain the action target of each compound, the species is selected as “homo sapiens”, and the screening condition is Probability> 0, so as to obtain effective target, and use UniProt database for target gene annotation conversion.

[0014] As a further scheme of the application: the specific steps of step two are as follows: first, search related targets in GeneCards, OMIM, DrugBank and TTD databases with “Heart failure with preserved ejection fraction” as the keyword, further after summarizing and de-duplicating, obtain the potential action target of heart failure with preserved ejection fraction.

[0015] As a further scheme of the application: the specific steps of step three are as follows: first, use online Wayne diagram tool to analyze the intersection target of effective component target and heart failure with preserved ejection fraction potential target, further import the intersection target into String database for protein-protein interaction (PPI) analysis, remove independent target, and set the highest confidence as 0.9 as the condition for screening drug anti-heart failure with preserved ejection fraction core target, obtain tsv file, further import tsv file into Cytoscape3.7.2 software for PPI network visualization processing, and screen out target points above the median value in degree value sorting as core target points.

[0016] As a further scheme of the present application: the specific steps of step four are as follows: first, the core target points obtained by screening in step three are introduced into the Metascape database for KEGG pathway analysis and GO enrichment analysis, then the top 10 and top 20 results are taken respectively to draw GO column chart and KEGG bubble pathway chart, and R software is used for result visualization.

[0017] Compared with the prior art, the beneficial effects of the present application are:

[0018] 1. The present application preliminarily predicts the key target points and signal pathways of Qihong Capsule in treating HFpEF by using network pharmacology method, and further verifies the clinical efficacy and potential mechanism based on the results, thereby providing more objective basis for Qihong Capsule in treating HFpEF, and laying a foundation for the in-depth study of the treatment of HFpEF by the method of tonifying qi and promoting blood circulation.

[0019] 2. The present application discusses the efficacy and mechanism of Qihong Capsule on HFpEF by the method of combination of network pharmacology and clinical verification, and the results show that Qihong Capsule can regulate cGMP / PKG signal pathway and multiple inflammation-related pathways, and has good treatment potential in improving the cardiac function of HFpEF patients, thereby providing a scientific basis for the clinical promotion and modern application of traditional Chinese medicine. BRIEF DESCRIPTION OF DRAWINGS

[0020] Fig. 1 Wayne diagram for Qihong Capsule and related target points of heart failure with preserved ejection fraction.

[0021] Fig. 2 PPI network diagram for common target points.

[0022] Fig. 3 GO enrichment analysis result graph (1).

[0023] Fig. 4 GO enrichment analysis result graph (2).

[0024] Fig. 5 GO enrichment analysis result graph (3).

[0025] Fig. 6 KEGG pathway enrichment analysis bubble chart. DETAILED DESCRIPTION

[0026] The technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0027] Referring to Figs. 1-6 In the embodiments of the present application, a network pharmacology-based analysis method for treating heart failure with preserved ejection fraction by using Qihong capsules is provided, and the method steps are as follows:

[0028] Step 1: Screening of effective components of Qihong capsules and core target points

[0029] The compounds of Astragalus, Ramulus Cinnamomi, Rhodiolae, Salviae Miltiorrhizae, Alismatis, and Semen Lepidii were collected and sorted from TCMSP database (http: / / tcmspw.com / tcmsp.php), China National Knowledge Infrastructure (https: / / www.cnki.net), Wanfang Data (https: / / www.wanfangdata.com.cn / ), PubMed (https: / / pubmed.ncbi.nlm.nih.gov / ), and other related literatures. The ADMET parameters of these compounds were queried from TCMSP (http: / / tcmspw.com / tcmsp.php) and admetSAR (http: / / lmmd.ecust.edu.cn / ) databases, and the effective compound components with oral bioavailability (OB) ≥ 30% and drug-likeness (DL) ≥ 0.18 were screened. Subsequently, the SMILES structure of these effective compounds was found by using PubChem (https: / / pubchem.ncbi.nlm.nih.gov / ) website, and was imported into SWISSTargetPrediction database (http: / / www.swisstargetprediction.ch) to obtain the action target point of each compound. The species was selected as “homo sapiens”, and the screening condition was Probability>0, so as to obtain the effective target point, and the target point gene annotation conversion was performed by using UniProt database (https: / / www.uniprot.org / ).

[0030] Step 2: Obtaining of heart failure with preserved ejection fraction gene target points

[0031] The related targets were retrieved in databases such as GeneCards (https: / / www.genecards.org), OMIM (https: / / db.idrblab.org / ttd / ), DrugBank (https: / / www.drugbank.ca) and TTD (http: / / db.idrblab.net / ttd) with “Heart failure with preserved ejection fraction” as the keyword. After summarizing and deduplication, the potential action targets of heart failure with preserved ejection fraction were obtained.

[0032] Step three: Construction of the protein-protein interaction (PPI) network of Qihong capsule-HFpEF protein

[0033] The intersection targets of the active ingredient targets and the potential targets of heart failure with preserved ejection fraction were analyzed by using the online Venn diagram tool (http: / / bioinformatics.psb.ugent.be / webtools / Venn / ). Then the intersection targets were imported into the String database (https: / / string-db.org) for protein-protein interaction (PPI) analysis. The independent targets were removed, and the highest confidence was set to 0.9 as the condition for screening the core targets of the drug against heart failure with preserved ejection fraction, and the tsv file was obtained. Next, the tsv file was imported into Cytoscape3.7.2 software for visualization processing of the PPI network, and the targets above the median value in the degree value sorting were selected as the core targets.

[0034] Step four: Gene ontology (GO) function enrichment analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis.

[0035] In order to clarify the role of the action targets of Qihong capsule against heart failure with preserved ejection fraction in the pathway, the core targets screened in step three were imported into the Metascape database (https: / / metascape.org / gp / index.html) for KEGG pathway analysis and GO enrichment analysis. The top 10 and top 20 results were taken to draw GO column chart and KEGG bubble pathway chart respectively, and R software was used for visualization of the results.

[0036] Clinical verification

[0037] 1. Selection and grouping of research subjects;

[0038] The present application selects 70 patients who are diagnosed as HFpEF and treated in the Department of Cardiology of the Traditional Chinese Medicine Hospital of Xinjiang Uygur Autonomous Region from June 2023 to June 2024. The inclusion criteria are as follows: meeting the diagnostic criteria for preserved ejection fraction heart failure in the Chinese Heart Failure Diagnosis and Treatment Guidelines 2018; age 30-80 years old, gender unrestricted; voluntarily participating in the study and signing the informed consent form. The exclusion criteria are as follows: uncontrolled myocarditis, left ventricular outflow tract obstruction, aneurysm, unstable angina pectoris, acute myocardial infarction; planned heart valve, vascular surgery or cardiac resynchronization therapy within 12 weeks; severe peripheral arterial disease, acute exacerbation of chronic obstructive pulmonary disease, pulmonary vascular disease; severe liver disease, kidney disease, blood system disease or systemic disease; uncontrolled hypertension (systolic blood pressure ≥180 mmHg or diastolic blood pressure ≥110 mmHg); pregnant or lactating women; known or suspected allergy to study drugs; received other drug clinical trials within 30 days. The included patients signed the informed consent form (ethical approval number: 2023XE-YS0012). The patients were randomly divided into Qihong capsule group and control group, 35 cases in each group. The patients in the control group received conventional treatment, including furosemide tablets, metoprolol sustained-release tablets, spironolactone tablets and sacubitril valsartan tablets. The patients in the Qihong capsule group took Qihong capsule on the basis of conventional treatment, 4 capsules each time, 3 times a day, orally, for 12 weeks of continuous treatment;

[0039] 2. Assessment of cardiac function

[0040] Before and after treatment, the serum NT-proBNP value and the left ventricular diastolic early and late blood flow velocity ratio (E / A value) and the left ventricular diastolic early blood flow velocity to mitral annulus diastolic early movement velocity ratio (E / e' value) detected by echocardiography were used to evaluate the improvement of the patient's cardiac function. The change of serum NT-proBNP concentration was detected by high sensitivity chemiluminescence immunoassay (CLIA). The E / A ratio represents the peak ratio of the early diastolic wave (E wave) to the late diastolic wave (A wave) in the left ventricular diastolic period, which is used to evaluate the diastolic function of the left ventricle. When the E / A ratio is normal, it indicates that the cardiac diastolic function is good; when it is lower than the normal range, it may indicate cardiac diastolic dysfunction. The E / e' ratio is the ratio of left ventricular diastolic early blood flow velocity to mitral annulus diastolic early movement velocity, which can also be used to evaluate the diastolic function of the left ventricle;

[0041] 3. Determination of serum inflammatory factors and cGMP levels

[0042] Before and after treatment, the patients' elbow venous blood samples were collected using standardized blood sampling procedures. All blood samples were immediately sent to the laboratory for serum separation and stored in a -80°C refrigerator. After the blood sample collection was completed, the levels of IL-1β, IL-6 and cGMP in the serum were determined by enzyme-linked immunosorbent assay (ELISA).

[0043] Statistical analysis

[0044] All data were statistically analyzed using SPSS 25.0 software. Measurement data were expressed as mean ± standard deviation, and the differences between groups were compared using t test. If the data met the normal distribution and homogeneity of variance, one-way analysis of variance was used for comparison between groups, and LSD method was used for multiple comparisons; if the data did not meet the homogeneity of variance, Tamhane T2 test was used. P<0.05 was considered statistically significant.

[0045] Results

[0046] Network pharmacology analysis

[0047] 1. Screening of effective components and target points of Qihong capsule and HFpEF

[0048] Through screening, 82 effective components and 877 effective target points of Qihong capsule were obtained. Through database retrieval, 415 HFpEF related target points were obtained. The intersection of Qihong capsule effective target points and HFpEF related target points was obtained, and 108 common target points were obtained, that is, the target points of Qihong capsule acting on HFpEF (such as Fig. 1 );

[0049] 2. Construction of PPI network diagram and acquisition of core target points

[0050] The target point interaction results were imported into Cytoscape software to construct the drug-disease target point interaction PPI network diagram (such as Fig. 2). After screening by STRING database analysis, 15 core targets were obtained: tumor protein p53 (TP53), JUN proto-oncogene (JUN), serine / threonine-protein kinase AKT1 (AKT1), tumor necrosis factor (TNF), interleukin 6 (IL-6), interleukin-1 beta (IL-1β), mitogen-activated protein kinase 1 (MAPK1), B-cell lymphoma-2 (BCL-2), C-X-C motif chemokine ligand 8 (CXCL8), C-C motif chemokine ligand 2 (CCL2), recombinant V-Rel reticuloendotheliosis viral oncogene homolog A (RELA), cysteine aspartate-specific protease 3 (CASP3), interferon gamma (IFNG), cyclin D1 (CCND1), and epidermal growth factor receptor (EGFR). Analysis of the molecular mechanisms of the above core targets found that each target played an important role in different biological processes, especially in inflammatory injury, cell proliferation, or apoptosis mechanisms;

[0051] 3. GO function and KEGG pathway enrichment analysis of Qihong capsule and HFpEF

[0052] GO function annotation contains 366 entries, including 297 biological processes (BP), 59 cell components (CC), and 10 molecular functions (MF). Based on the results of the corrected P-value ranking, the top 10 of each were selected and displayed in a bar chart (such as Figs. 3-5, Table 1). This shows that Qihong capsule mainly regulates the biological processes of vasodilation, inflammatory response, regulation of vascular contraction, positive regulation of MAPK cascade, response to hypoxia, etc.; in terms of cell components, it is mainly involved in functional locations such as plasma membrane, extracellular region, and membrane raft; in terms of molecular function, it is mainly related to endopeptidase activity, serine-type endopeptidase activity, metalloendopeptidase activity, peptidase activity, and protein serine / threonine / tyrosine kinase activity;

[0053] Table 1 GO enrichment analysis results of core target points

[0054]

[0055] KEGG enrichment analysis identified 113 signaling pathways. According to the sorted P value after correction, the top 20 pathways were selected for bubble chart (such as Fig. 6 , Table 2). The main pathways involved are cGMP-PKG signaling pathway, AGE-RAGE signaling pathway, lipid and atherosclerosis pathway, neuroactive ligand-receptor interaction related pathway, and cAMP signaling pathway. The cGMP / PKG signaling pathway has the most enriched target points;

[0056] Table 2 Top 20 pathways of KeGG pathway analysis

[0057] Clinical efficacy of Qihong capsule in the treatment of HFpEF

[0058] 1. General information

[0059] Seventy patients with HFpEF who visited the Department of Cardiology of the Traditional Chinese Medicine Hospital of Xinjiang Uygur Autonomous Region from June 2023 to June 2024 were selected, and all of them signed the informed consent form. According to the random number table method, they were divided into Qihong capsule group and control group (35 cases in each group). There was no statistically significant difference in general information between the two groups (P>0.05), and they were comparable. The Qihong capsule group was (64.57±11.47) years old, with 8 males and 27 females. The control group was (60.81±10.83) years old, with 7 males and 28 females.

[0060] 2. Effect of Qihong capsule on NT-proBNP level, E / A value and E / e' value of patients with HFpEF

[0061] After 12 weeks of treatment, the NT-proBNP levels, E / A values and E / e' values of the patients in both groups were compared. The results showed that the NT-proBNP levels of the Qihong capsule group and the control group were statistically different (P<0.05), indicating that both groups improved after treatment. The E / A values and E / e' values before and after treatment were not statistically different (P>0.05), and the difference between the Qihong capsule group and the control group before and after treatment was also not statistically different (P>0.05), but there was a trend of improvement (Table 3);

[0062] Table 3 Comparison of NT-proBNP, E / A values and E / e' values of patients in both groups before and after treatment

[0063] Note: * indicates P<0.05 before and after treatment

[0064] 3. Effect of Qihong capsule on serum IL-1β, IL-6 and cGMP of HFpEF patients;

[0065] Before treatment, the serum IL-1β, IL-6 and cGMP levels of the patients in both groups were not significantly different (P>0.05). After treatment, the IL-1β and IL-6 levels of the Qihong capsule group were significantly reduced, and the difference was statistically significant. The change in IL-1β of the control group was statistically different, while the change in IL-6 was not statistically different. In addition, the cGMP level of the Qihong capsule group increased from (1.22±0.26) nmol / L to (2.27±0.45) nmol / L (P<0.01), while the cGMP level of the control group increased from (1.25±0.32) nmol / L to (1.91±0.47) nmol / L (P<0.01), which was statistically different (P<0.05), but the change was not as significant as the Qihong capsule group. This indicates that Qihong capsule may have an activating effect on the cGMP / PKG pathway and can improve the inflammatory response of blood vessels, as shown in Table 4;

[0066] Table 4 Comparison of IL-1β, IL-6 and cGMP of patients in both groups before and after treatment

[0067]

[0068]

[0069] Note: * indicates P<0.05 before and after treatment

[0070] Conclusion

[0071] 1. Study on the pathogenesis of HFpEF;

[0072] HFpEF accounts for about 50% of all heart failure patients, but due to the unclear underlying pathogenesis, there is still a lack of evidence-based therapy [9]. Studies have shown that the pathogenesis of HFpEF may be related to abnormal cardiac systolic function, pulmonary hypertension and right ventricular dysfunction, cardiac lipotoxicity, coronary microcirculation disorder, skeletal muscle metabolism, myocardial inflammatory injury, etc. in addition to ventricular diastolic dysfunction

[10] . Cardiac inflammation stimulation may be one of the important pathological mechanisms of HFpEF, which can reduce the bioavailability of myocardial NO, affect the conduction of NO / sGC / cGMP signaling pathway, reduce the activation of PKG, thereby reducing the phosphorylation of sarcomere protein Titin, increasing myocardial passive stiffness, and reducing cardiac diastolic function

[11] . The GMP / PKG signaling pathway can be activated through two intrinsic pathways of NO soluble guanylate cyclase (NO-sGC) or natriuretic peptide-microgranular guanylate cyclase (NP-pGC), which can inhibit myocardial hypertrophy and ventricular hypertrophy, regulate cardiac remodeling, and thus improve the cardiac function of HFpEF patients [12-14]. Given the impaired cGMP-PKG signaling in HFpEF, activation of these pathways may be an effective strategy for treating HFpEF patients.

[0073] 2. Prevention and treatment of HFpEF by traditional Chinese medicine and characteristics of Qihong Capsule;

[0074] HFpEF, according to the clinical manifestations of TCM, is classified into the category of heart water, edema, chest stuffiness and other syndromes. TCM believes that HFpEF is mainly in the heart, with deficiency in the heart, spleen and kidney, and excess in phlegm turbidity and blood stasis. Compared with heart failure with reduced ejection fraction, the overall pathological changes are mainly excess, with phlegm and blood stasis as the main pathological factors, and deficiency in qi, with less involvement of yang deficiency, and mild degree of deficiency syndrome. The pathogenesis is related to qi and stasis, with common symptoms of heart qi deficiency and qi deficiency and blood stasis. The basic pathogenesis of HFpEF, deficiency and excess, determines the general treatment principle of strengthening the body resistance and eliminating evil. Qihong Capsule, a representative formula of tonifying qi and promoting blood circulation, is a Chinese medicine protocol developed based on the theory of treating phlegm and blood stasis together and modern TCM theory for the treatment of chronic heart failure. Its composition is: Huangqi 15g, Guizhi 9g, Hongjingtian 9g, Danshen 9g, Zexie 9g, Tingshili 12g, with the functions of tonifying qi and promoting blood circulation, warming yang and unblocking collaterals. Huangqi is the monarch in the formula, with sweet and warm properties, and entering the spleen and lung meridians, which can tonify qi and ascend yang, reinforce the defensive aspect and consolidate the exterior. Guizhi and Hongjingtian are the ministers, with pungent and sweet flavors, warm property, and entering the heart, lung and bladder meridians, which can sweat and disperse the exterior, warm the meridians and assist yang and qi. Huangqi can warm yang and promote blood circulation, and Guizhi can strengthen the function of warming the meridians. Hongjingtian has the functions of tonifying qi and blood, strengthening the body resistance, promoting blood circulation and stopping bleeding. Danshen, Tingshili and Zexie are the ministers, which can tonify the heart and lung, and the kidney essence, tonify the blood of the heart and liver, promote blood circulation and remove blood stasis, and tonify without stagnation. The combination of these herbs can tonify qi for the root and remove phlegm and blood stasis, promote blood circulation and remove blood stasis for the branch, with tonifying and removing phlegm and blood stasis, and promoting blood circulation and removing blood stasis, which can treat both the root and branch, and tonify qi and promote blood circulation and warm yang and unblock collaterals.

[0075] 3. Network pharmacology predicts effective targets of Qihong Capsule for the treatment of HFpEF;

[0076] Network pharmacology is the integration of drug action network and biological network to analyze the interaction between drugs and nodes or network modules in the network, from single target to comprehensive network analysis. In recent years, some studies have used network pharmacology to explore the effective targets of Yiqi Huoxue Chinese medicine in the treatment of heart failure. For example, Zhao et al. proved that Qili Qiangxin capsule may regulate INS, ALB and other related targets through cGMP / PKG signaling pathway and calcium signaling pathway, and play a role in the treatment of chronic heart failure

[20] . Chen et al. found that Qishen Yixin granules may act on TNF, AKT1, IL-6, P53, JUN and other targets, and regulate gene expression, cell apoptosis and proliferation, hypoxia response, and inflammatory response to treat HFpEF

[21] . Hao et al. found that Qili Qiangxin capsule may also regulate cGMP-PKG signaling pathway to treat HFpEF

[22] . In this study, we used network pharmacology to analyze the potential role of Qihong capsule in improving heart function and regulating inflammation-related molecular signaling pathways in patients with HFpEF. We found that the active ingredients of Qihong capsule can act on up to 108 HFpEF-related targets, among which core targets such as TP53, JUN, AKT1 are closely related to key biological processes such as inflammation and apoptosis. GO function enrichment analysis showed that the targets of Qihong capsule were mainly involved in blood circulation, heart contraction and other biological processes. KEGG pathway analysis revealed that Qihong capsule may act through cGMP / PKG signaling pathway, calcium signaling pathway, AGE-RAGE signaling pathway and inflammation-related MAPK pathway. Therefore, cGMP / PKG signaling pathway may be an important regulatory pathway for Yiqi Huoxue Chinese medicine in the treatment of heart failure with preserved ejection fraction.

[0077] 4. Verification of clinical efficacy of Qihong powder in the treatment of HFpEF and exploration of its mechanism

[0078] Previous studies have confirmed that Qihong capsule can reduce the NT-proBNP level of patients with HFpEF, improve the NYHA cardiac function classification, increase the distance of 6MWT, and significantly improve the quality of life of patients, with significant efficacy and good safety. In addition, studies have shown that Qihong capsule can also improve the clinical symptoms of patients with HFrEF, improve the physical fitness and quality of life of patients, increase their ejection fraction, and reduce the NT-proBNP level. To further verify the effect of Qihong capsule on HFpEF and its mechanism, this study focuses on observing the NT-proBNP value and echocardiogram (E / A value and E / e' value) before and after treatment to evaluate the cardiac function and diastolic function of patients. The NT-proBNP value is a biomarker that reflects the cardiac load and cardiac function failure, and by detecting the change in its serum concentration, the improvement of the cardiac function of patients can be directly observed. The E / A value and E / e' value in echocardiogram are important parameters for evaluating the diastolic function of the heart, the former reflects the diastolic filling capacity of the left atrium and left ventricle, and the latter is related to the left ventricular filling pressure and diastolic pressure. In this study, we confirmed that the NT-proBNP level of patients in the Qihong capsule group was significantly reduced, which was consistent with previous studies, and the E / A value and E / e' value had an improvement trend, indicating that the cardiac function of patients improved and the diastolic function of the heart also improved to a certain extent. Previous studies have shown that Qihong capsule can increase coronary blood flow and venous oxygen content, thereby improving cardiac function, and Qihong capsule can also increase the ejection fraction and short-axis shortening rate of myocardial ischemia-induced chronic heart failure model rats, and has a protective effect. In this study, we further studied the effect of Qihong capsule on the inflammatory damage of HFpEF, and the results showed that the serum IL-1β and IL-6 levels of patients in the Qihong capsule group were significantly decreased, and the cGMP level was significantly increased, indicating that Qihong capsule may reverse cardiac function damage and improve the cardiac function of HFpEF patients by inhibiting inflammatory response and activating the cGMP / PKG pathway, which further supports the efficacy of Qihong capsule on HFpEF.

[0079] 5. Future Prospects

[0080] This study explores the efficacy of Qihong capsule on HFpEF and its mechanism of action through the combination of network pharmacology and clinical verification. The results show that Qihong capsule may regulate the cGMP / PKG signaling pathway and multiple inflammation-related pathways, and has good therapeutic potential in improving the cardiac function of HFpEF patients. This provides a scientific basis for the clinical promotion and modern application of traditional Chinese medicine, and in future research, the internal mechanism of action will be further explored around this target.

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[0108] Although the present application has been described in detail with reference to the foregoing embodiments, those skilled in the art can modify the technical solutions described in the foregoing embodiments, or make equivalent replacements to part of the technical features, and any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. A method for analyzing the treatment of preserved ejection fraction heart failure by Qihong Capsules based on network pharmacology, characterized by: The method steps are as follows: Step one: effective components of Qihong capsule and core target screening; Step two: obtaining heart failure with preserved ejection fraction gene target; Step three: Qihong capsule-HFpEF protein-protein interaction network construction; Step four: gene ontology function enrichment analysis and kyoto gene and genome encyclopedia pathway enrichment analysis.

2. The network pharmacology-based analysis method for treating heart failure with preserved ejection fraction with Qihong Capsules according to claim 1, characterized in that: The specific steps of step one are as follows: first, collect and organize the compounds of Astragalus, Ramulus Cinnamomi, Rhodiolae, Salviae Miltiorrhizae, Alismatis, and Semen Lepidii from TCMSP database, China National Knowledge Network, Wanfang Data, and PubMed literature, further query the ADME parameters of these compounds through TCMSP and admetSAR databases, screen effective compound components with oral bioavailability (OB) ≥ 30% and drug-like property (DL) ≥ 0.18, then find the SMILES structure of these effective compounds through PubChem website, and import them into SWISS TargetPrediction database to obtain the action target of each compound, with the species selected as "homo sapiens" and the screening condition as Probability > 0, so as to obtain effective targets, and use UniProt database for target gene annotation conversion. 3.The network pharmacology-based analysis method for treating heart failure with preserved ejection fraction by using Qihong Capsules according to claim 1, characterized in that: The specific steps of step two are as follows: first, search related targets in GeneCards, OMIM, DrugBank, and TTD databases with "Heart failure with preserved ejection fraction” as the keyword, further obtain the potential action targets of heart failure with preserved ejection fraction after summarizing and de-duplicating.

4. The network pharmacology-based analysis method for treating heart failure with preserved ejection fraction with Qihong capsule according to claim 1, wherein the specific steps of step three are as follows: first, analyze the intersection targets of effective component targets and potential targets of heart failure with preserved ejection fraction by using online Wayne diagram tool, further import the intersection targets into String database for protein-protein interaction (PPI) analysis, remove independent targets, and set the highest confidence as 0.9 as the condition for screening drug anti-heart failure with preserved ejection fraction core targets, obtain tsv file, further import the tsv file into Cytoscape3.7.2 software for PPI network visualization processing, and screen out targets with degree value above the median in degree value sorting as core targets.

5. The network pharmacology-based analysis method for treating heart failure with preserved ejection fraction with Qihong capsule according to claim 1, wherein the specific steps of step four are as follows: first, import the core targets screened in step three into Metascape database for KEGG pathway analysis and GO enrichment analysis, further take the top 10 and top 20 results to draw GO column chart and KEGG bubble pathway chart respectively, and use R software for result visualization.

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