# Mechanism of Peitu Shengjin Formula Shenlingbaizhu Powder in Treating Bronchial Asthma and Allergic Colitis through Different Diseases with Simultaneous Treatment Based on Network Pharmacology and Molecular Docking

Authors: Liying Zeng; Shaodan Sun; Peiwen Chen; Qina Ye; Xiaoling Lin; Hongjun Wan; Yawen Cai; Xiaogang Chen
Journal: Evidence-Based Complementary and Alternative Medicine
 (2022)

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## Abstract

Background. Shenlingbaizhu powder (SLBZP), one of the classic Earth-cultivating and gold-generating prescriptions of traditional Chinese medicine, is widely used to treat various diseases. However, the pharmacological mechanisms of SLBZP on bronchial asthma (BA) and allergic colitis (AC) remain to be elucidated. Methods. Network pharmacology and molecular docking technology were used to explore the potential mechanism of SLBZP in treating BA and AC with the simultaneous treatment of different diseases. The potential active compounds of SLBZP and their corresponding targets were obtained from BATMAN-TCM, ETCM, SymMap TCM@TAIWAN, and TCMSP databases. BA and AC disease targets were collected through DisGeNET, TTD, GeneCards, PharmGKB, OMIM, NCBI, The Human Phenotype Ontology, and DrugBank databases. Common targets for drugs and diseases were screened by using the bioinformatics and evolutionary genomics platform. The analyses and visualizations of Gene Ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment of common targets were carried out by R software. The key targets were screened by using the plug-in “cytoHubba” of Cytoscape software, and the “active compound-key target” network was constructed. Molecular docking analysis was performed using AutoDock software. The miRTarBase database was used to predict microRNAs (miRNAs) targeting key targets, and the key target-miRNA network was constructed. Result. Through screening, 246 active compounds and 281 corresponding targets were obtained. Common targets were mainly enriched in 2933 biological processes and 182 signal pathways to play the role of treating BA and AC. There were 131 active compounds related to key targets. The results of molecular docking showed that the important active compounds in SLBZP had good binding ability with the key targets. The key target-miRNA network showed that 94 miRNAs were predicted. Conclusion. SLBZP has played the role of treating different diseases with the same treatment on BA and AC through the characteristics of multicompound, multitarget, and multipathway of traditional Chinese medicine, which provides a theoretical basis for explaining the mechanism and clinical application of SLBZP treating different diseases with the same treatment in BA and AC.


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## Body

## 1. Introduction

Asthma generally refers to bronchial asthma (BA). BA, one of the most common chronic noncommunicable diseases in children and adults, is characterized by variable respiratory symptoms and variable airflow limitation, which is the result of complex gene-environment interactions, and is heterogeneous in clinical manifestations and the type and intensity of airway inflammation and remodeling [1]. The goal of BA treatment is to achieve good asthma control, that is, to minimize the burden of symptoms and the risk of deterioration [2]. However, asthma attacks and hospitalizations are frequent, and the mortality rate remains high. Strategies need to be developed to change the natural history of BA and prevent serious deterioration and the decline of lung function [1]. Allergic colitis (AC), an inflammatory disease, is characterized by the infiltration of eosinophils into the colon wall and the presence of red blood in the stool of healthy breast-fed or formula-fed infants, which usually develops in the first few weeks or months of life and can be a benign and/or severe disease in infant gastrointestinal diseases [3–4]. To date, the most effective interventions are preventive methods, especially feeding strategies, to reduce the incidence of disease while establishing adequate growth and progression to enteral feeding [5]. However, their pathogenesis has not yet been fully clarified with some allergens unclear or unavoidable, and modern medicine lacks ideal preventive and therapeutic methods [6]. At present, modern medicine adopts allergen avoidance, desensitization, and symptomatic treatment, but some antihistamines and antileukotrienes need to be taken for a long time, which brings certain economic burden and psychological impact to patients and cannot completely cure allergic diseases with some deficiencies, such as side effects of drugs and easy recurrence after withdrawal [7–9]. In recent years, treating allergic diseases with traditional Chinese medicine has been more and more widely used in clinical practice with various methods, remarkable effects, less adverse reactions in long-term application, and good compliance, which is convenient for clinical promotion [10, 11].Shelingbaizhu powder (SLBZP), from the Prescriptionsof Peaceful Benevolent Dispensary and composed of 10 Chinese medicines including renshen (Panax ginseng C. A. Mey.), fuling (Poria cocos (Schw.) Wolf.), baizhu (Atractylodes macrocephala Koidz.), baibiandou (Lablab Semen Album), shanyao (Rhizoma Dioscoreae), lianzi (Semen Nelumbinis), yiyiren (Coicis Semen), sharen (Amomum aurantiacum H. T. Tsai Et S. W. Zhao), jiegeng (Platycodon grandiforus), and gancao (licorice), has the effects of replenishing qi, strengthening spleen, excreting dampness, and stopping diarrhea [12]. Previous studies have shown that SLBZP can regulate intestinal water metabolism and intestinal flora, inhibit inflammatory response, repair intestinal mucosal barrier, and enhance colonic motility, which is widely used in the clinical treatment of ulcerative colitis, chronic diarrhea, chronic obstructive pulmonary disease, bronchial asthma, diabetes, eczema, allergic rhinitis, etc. [13, 14].Network pharmacology, targeting biological networks, analyzes the connections between drugs, targets, and diseases in these networks. A comprehensive and systematic research on network pharmacology conforms to a holistic view, which is the main characteristic of many traditional medicines. Studies have shown that many traditional medicines exhibit synergistic effects by acting on multiple targets and pathways at different levels through network pharmacology [15]. This method effectively bridges the gap between modern medicine and traditional medicine and greatly promotes the research on the synergy of traditional medicine. Different diseases with simultaneous treatment means that the same pathogenesis appears in the occurrence and development of different diseases, and the same treatment can be adopted. SLBZP reinforces Earth to generate metal for treating BA and AC, which is in line with the concept of different diseases with simultaneous treatment. This study comprehensively analyzed and explored the mechanism of SLBZP in treating BA and AC with simultaneous treatment of different diseases from compounds, targets, pathways, biological processes, etc., by network pharmacology and molecular docking, which conforms to the overall function of traditional Chinese medicine theory and provides theoretical bases for clarifying the action mechanism of SLBZP on BA and AC and promoting its clinical application (Figure 1).Figure 1
Workflow for exploring the mechanisms of Shenlingbaizhu powder in treating bronchial asthma and allergic colitis with simultaneous treatment of different diseases.


## 2. Materials and Methods

### 2.1. Screening Compounds and Targets of SLBZP

The active compounds of SLBZP were separately obtained from these databases: BATMAN-TCM (http://bionet.ncpsb.org.cn/batman-tcm/index.php/Home/Index/index) [16], ETCM (http://www.tcmip.cn/ETCM/index.php/Home/Index/) [17], SymMap (http://www.symmap.org/) [18] and Traditional Chinese Medicine Database@TAIWAN (http://tcm.cmu.edu.tw/review.php?menuid=3) [19]. Then, the active compounds that had good oral bioavailability (OB) and drug similarity (DL) and their targets of SLBZP were screened out under the conditions of OB ≥ 30% and DL ≥ 0.18 by entering the above obtained active compounds into Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP, http://lsp.nwu.edu.cn/tcmsp.php) [20]. Meanwhile, the active compounds and their targets of SLBZP from the TCMSP database were also obtained with OB ≥ 30% and DL ≥ 0.18. Next, all these obtained active compounds were synthesized to remove duplications. The full names of the targets screened by TCMSP were input into the DrugBank database (https://www.drugbank.ca/) [21] and UniProt database (https://www.uniprot.org/?tdsourcetag=s_pcqq_aiomsg) [22] to get the gene symbol and UniProt ID, which were all standardized and normalized to ensure accuracy.


### 2.2. Screening Targets of BA and AC

The target genes related to BA were obtained with the keyword “bronchial asthma” and the species set as “Homo sapiens” from these 8 databases: DisGeNET (http://www.disgenet.org/web/DisGeNET/menu/search) [23], TTD (https://db.idrblab.org/ttd/) [24], GeneCards (https://www.genecards.org) [25], PharmGKB (https://www.pharmgkb.org/) [26], OMIM (https://omim.org/) [27], NCBI (https://www.ncbi.nlm.nih.gov/gene) [28], The Human Phenotype Ontology (https://hpo.jax.org/app/) [29], and DrugBank. The target genes related to AC were obtained with the keyword “allergic colitis” and the species set as “Homo sapiens” from these 5 databases: TTD, GeneCards, PharmGKB, OMIM, and NCBI. The obtained data were combined separately, and then the duplications were removed. The full name of the last screened target genes were input into the DrugBank database and UniProt database to get the gene symbol and UniProt ID, which were also all standardized and normalized to ensure accuracy.


### 2.3. Screening of Common Targets

The targets related to active compounds, BA, and AC were matched and mapped by using the bioinformatics and evolutionary genomics platform (http://bioinformatics.psb.ugent.be/webtools/Venn/). At the same time, a Venn diagram was drawn to obtain the common targets of the active compounds of SLBZP for treating BA and AC.


### 2.4. GO and KEGG Enrichment Analysis of Common Targets

The enrichment analysis and visualization of Gene Ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were carried out for the common targets of SLBZP in treating BA and AC with the species set as “Homo sapiens” and the threshold set asP < 0.05 by the “ggplot2”, “enrichplot”, “clusterprofiler” [30], and “ggpubr” packages of R software (version 3.6.1).


### 2.5. Construction of Active Compound-Key Target Network

The obtained common targets were imported into Cytoscape software (version 3.8.0;http://www.cytoscape.org) [31], and the “cytoHubba” plug-in was used to screen out the key targets. Then, an active compound-key target network was constructed by Cytoscape software, of which the network topology analysis was carried out by “Network Analysis” in the tool. The network showed the connection between the active compounds and key targets, and the molecular mechanism of SLBZP in treating BA and AC was explored on this basis.


### 2.6. Molecular Docking Verification

According to the above analysis results, the key target proteins and the important active compounds were molecularly docked. The protein structures of the targets were obtained from the RCSB PDB database (https://www.rcsb.org/) [32]. The 2D structures of the active compounds were obtained from the PubChem database (https://pubchem.ncbi.nlm.nih.gov/) [33] and were optimized to save as 3D structures with Chem3D software. AutoDockTools and AutoDockVina software were used for molecular structure processing and molecular docking. PyMOL and Discovery Studio were used to visualize the docking results.


### 2.7. Construction of Key Target-microRNA (miRNA) Network

The miRTarBase database (https://mirtarbase.cuhk.edu.cn/%7EmiRTarBase/miRTarBase_2019/php/index.php) is used to predict upstream miRNAs targeting key targets [34]. The collected miRNA-mRNA interactions have been verified by different types of experiments including report analyses in miRTarBase, western blot, qPCR, microarray, and next-generation sequencing experiments. In order to make predictions more reliable and accurate, only miRNAs that may interact with the targets were obtained through reporter gene analyses. After selecting “By Target Gene” and the species as “Human”, key targets were entered to predict miRNAs. Then, the key targets and their corresponding predicted miRNAs were organized into an Excel file that was imported into Cytoscape software. Finally, the network of the predicted miRNAs and key targets were constructed by Cytoscape software.


## 2.1. Screening Compounds and Targets of SLBZP

The active compounds of SLBZP were separately obtained from these databases: BATMAN-TCM (http://bionet.ncpsb.org.cn/batman-tcm/index.php/Home/Index/index) [16], ETCM (http://www.tcmip.cn/ETCM/index.php/Home/Index/) [17], SymMap (http://www.symmap.org/) [18] and Traditional Chinese Medicine Database@TAIWAN (http://tcm.cmu.edu.tw/review.php?menuid=3) [19]. Then, the active compounds that had good oral bioavailability (OB) and drug similarity (DL) and their targets of SLBZP were screened out under the conditions of OB ≥ 30% and DL ≥ 0.18 by entering the above obtained active compounds into Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP, http://lsp.nwu.edu.cn/tcmsp.php) [20]. Meanwhile, the active compounds and their targets of SLBZP from the TCMSP database were also obtained with OB ≥ 30% and DL ≥ 0.18. Next, all these obtained active compounds were synthesized to remove duplications. The full names of the targets screened by TCMSP were input into the DrugBank database (https://www.drugbank.ca/) [21] and UniProt database (https://www.uniprot.org/?tdsourcetag=s_pcqq_aiomsg) [22] to get the gene symbol and UniProt ID, which were all standardized and normalized to ensure accuracy.


## 2.2. Screening Targets of BA and AC

The target genes related to BA were obtained with the keyword “bronchial asthma” and the species set as “Homo sapiens” from these 8 databases: DisGeNET (http://www.disgenet.org/web/DisGeNET/menu/search) [23], TTD (https://db.idrblab.org/ttd/) [24], GeneCards (https://www.genecards.org) [25], PharmGKB (https://www.pharmgkb.org/) [26], OMIM (https://omim.org/) [27], NCBI (https://www.ncbi.nlm.nih.gov/gene) [28], The Human Phenotype Ontology (https://hpo.jax.org/app/) [29], and DrugBank. The target genes related to AC were obtained with the keyword “allergic colitis” and the species set as “Homo sapiens” from these 5 databases: TTD, GeneCards, PharmGKB, OMIM, and NCBI. The obtained data were combined separately, and then the duplications were removed. The full name of the last screened target genes were input into the DrugBank database and UniProt database to get the gene symbol and UniProt ID, which were also all standardized and normalized to ensure accuracy.


## 2.3. Screening of Common Targets

The targets related to active compounds, BA, and AC were matched and mapped by using the bioinformatics and evolutionary genomics platform (http://bioinformatics.psb.ugent.be/webtools/Venn/). At the same time, a Venn diagram was drawn to obtain the common targets of the active compounds of SLBZP for treating BA and AC.


## 2.4. GO and KEGG Enrichment Analysis of Common Targets

The enrichment analysis and visualization of Gene Ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were carried out for the common targets of SLBZP in treating BA and AC with the species set as “Homo sapiens” and the threshold set asP < 0.05 by the “ggplot2”, “enrichplot”, “clusterprofiler” [30], and “ggpubr” packages of R software (version 3.6.1).


## 2.5. Construction of Active Compound-Key Target Network

The obtained common targets were imported into Cytoscape software (version 3.8.0;http://www.cytoscape.org) [31], and the “cytoHubba” plug-in was used to screen out the key targets. Then, an active compound-key target network was constructed by Cytoscape software, of which the network topology analysis was carried out by “Network Analysis” in the tool. The network showed the connection between the active compounds and key targets, and the molecular mechanism of SLBZP in treating BA and AC was explored on this basis.


## 2.6. Molecular Docking Verification

According to the above analysis results, the key target proteins and the important active compounds were molecularly docked. The protein structures of the targets were obtained from the RCSB PDB database (https://www.rcsb.org/) [32]. The 2D structures of the active compounds were obtained from the PubChem database (https://pubchem.ncbi.nlm.nih.gov/) [33] and were optimized to save as 3D structures with Chem3D software. AutoDockTools and AutoDockVina software were used for molecular structure processing and molecular docking. PyMOL and Discovery Studio were used to visualize the docking results.


## 2.7. Construction of Key Target-microRNA (miRNA) Network

The miRTarBase database (https://mirtarbase.cuhk.edu.cn/%7EmiRTarBase/miRTarBase_2019/php/index.php) is used to predict upstream miRNAs targeting key targets [34]. The collected miRNA-mRNA interactions have been verified by different types of experiments including report analyses in miRTarBase, western blot, qPCR, microarray, and next-generation sequencing experiments. In order to make predictions more reliable and accurate, only miRNAs that may interact with the targets were obtained through reporter gene analyses. After selecting “By Target Gene” and the species as “Human”, key targets were entered to predict miRNAs. Then, the key targets and their corresponding predicted miRNAs were organized into an Excel file that was imported into Cytoscape software. Finally, the network of the predicted miRNAs and key targets were constructed by Cytoscape software.


## 3. Results

### 3.1. Acquirement of Active Compounds of SLBZP

Preliminarily, a total of 335 active compounds were acquired from the BATMAN-TCM database; a total of 443 active compounds were acquired from the ETCM database; a total of 1182 active compounds were acquired from the SymMap database; a total of 352 active compounds were acquired from the Traditional Chinese Medicine Database@TAIWAN database; and a total of 171 active compounds were acquired from the TCMSP database. At last, 217 eligible unique active compounds of SLBZP in total were retrieved from the TCMSP database under the conditions of OB ≥ 30% and DL ≥ 0.18, which are all shown in Table1.Table 1
Characteristics of eligible active compounds in SLBZP with OB and DL parameters.
CodeMolecule IDMolecule nameOB (%)DLHerbsP1MOL004924(-)-Medicocarpin40.990.95GancaoP2MOL004988Kanzonol F32.470.89GancaoP3MOL005018Xambioona54.850.87GancaoP4MOL005458Dioscoreside C_qt36.380.87ShanyaoP5MOL007536Stigmasta-5, 22-dien-3-beta-yl acetate46.440.86SharenP6MOL001474Sanguinarine37.810.86SharenP7MOL001973Sitosteryl acetate40.390.85SharenP8MOL004948Isoglycyrol44.70.84GancaoP9MOL008752Dihydroverticillatine42.690.84JiegengP10MOL000787Fumarine59.260.83RenshenP11MOL005357Gomisin B31.990.83RenshenP12MOL000300Dehydroeburicoic acid44.170.83FulingP13MOL000285(2R)-2-[(5R, 10S, 13R, 14R, 16R, 17R)-16-hydroxy-3-keto-4, 4, 10, 13, 14-pentamethyl-1, 2, 5, 6, 12, 15, 16, 17-octahydrocyclopenta[a]phenanthren-17-yl]-5-isopropyl-hex-5-enoic acid38.260.82FulingP14MOL000280(2R)-2-[(3S, 5R, 10S, 13R, 14R, 16R, 17R)-3, 16-dihydroxy-4, 4, 10, 13, 14-pentamethyl-2, 3, 5, 6, 12, 15, 16, 17-octahydro-1h-cyclopenta[a]phenanthren-17-yl]-5-isopropyl-hex-5-enoic acid31.070.82FulingP15MOL005317Deoxyharringtonine39.270.81RenshenP16MOL000283Ergosterol peroxide40.360.81FulingP17MOL0002873beta-hydroxy-24-methylene-8-lanostene-21-oic acid38.70.81FulingP18MOL0002767, 9(11)-Dehydropachymic acid35.110.81FulingP19MOL000289Pachymic acid33.630.81FulingP20MOL000546Diosgenin80.880.81ShanyaoP21MOL000275Trametenolic acid38.710.80FulingP22MOL005376Panaxadiol33.090.79RenshenP23MOL005401Ginsenoside Rg5_qt39.560.79RenshenP24MOL004917Glycyroside37.250.79GancaoP25MOL007535(5S, 8S, 9S, 10R, 13R, 14S, 17R)-17-[(1R, 4R)-4-ethyl-1, 5-dimethylhexyl]-10, 13-dimethyl-2, 4, 5, 7, 8, 9, 11, 12, 14, 15, 16, 17-dodecahydro-1h-cyclopenta[a]phenanthrene-3, 6-dione33.120.79SharenP26MOL005348Ginsenoside-Rh4_qt31.110.78RenshenP27MOL000033(3S, 8S, 9S, 10R, 13R, 14S, 17R)-10, 13-dimethyl-17-[(2R, 5S)-5-propan-2-yloctan-2-yl]-2, 3, 4, 7, 8, 9, 11, 12, 14, 15, 16, 17-dodecahydro-1h-cyclopenta[a]phenanthren-3-ol36.230.78BaizhuP28MOL009136Peraksine82.580.78FulingP29MOL000211Mairin55.380.78GancaoP30MOL005001Gancaonin H50.100.78GancaoP31MOL001323Sitosterol alpha143.280.78YiyirenP32MOL000279Cerevisterol37.960.77FulingP33MOL005465AIDS18090745.330.77ShanyaoP34MOL000449Stigmasterol43.830.76Renshen, yiyiren, sharen, baibiandou, shanyaoP35MOL000028α-Amyrin39.510.76BaizhuP36MOL000290Poricoic acid A30.610.76FulingP37MOL00175524-Ethylcholest-4-en-3-one36.080.76RenshenP38MOL004355Spinasterol42.980.76JiegengP39MOL004718α-Spinasterol42.980.76JiegengP40MOL005440Isofucosterol43.780.76ShanyaoP41MOL01062524-Methylenecholesterol43.540.76ShanyaoP42MOL000358Beta-sitosterol36.910.75Renshen, sharenP43MOL005399Alexandrin_qt36.910.75RenshenP44MOL001525Daucosterol36.910.75RenshenP45MOL000296Hederagenin36.910.75FulingP46MOL000292Poricoic acid C38.150.75FulingP47MOL000291Poricoic acid B30.520.75FulingP48MOL0063767-Dehydrosigmasterol37.420.75FulingP49MOL000359Sitosterol36.910.75Gancao, yiyirenP50MOL001771Poriferast-5-en-3beta-ol36.910.75SharenP51MOL013119Enhydrin40.560.74RenshenP52MOL000139Smitilbin37.600.74RenshenP53MOL009387Didehydrotuberostemonine51.910.74BaizhuP54MOL004903Liquiritin65.690.74GancaoP55MOL009154Tuberostemoenone53.900.73BaizhuP56MOL004891Shinpterocarpin80.300.73GancaoP57MOL009431Stemonine81.750.72BaizhuP58MOL000282Ergosta-7, 22e-dien-3beta-ol43.510.72FulingP59MOL009149Cheilanthifoline46.510.72FulingP60MOL004805(2S)-2-[4-hydroxy-3-(3-methylbut-2-enyl)phenyl]-8, 8-dimethyl-2, 3-dihydropyrano[2, 3-f]chromen-4-one31.790.72GancaoP61MOL00543524-Methylcholest-5-enyl-3belta-O-glucopyranoside_qt37.580.72ShanyaoP62MOL012254Campesterol37.580.71RenshenP63MOL005438Campesterol37.580.71Renshen, shanyaoP64MOL000493Campesterol37.580.71RenshenP65MOL00501318α-Hydroxyglycyrrhetic acid41.160.71GancaoP66MOL006070Robinin39.840.71JiegengP67MOL01104218Alpha-hydroglycyrrhetic acid38.930.71BaibiandouP68MOL004567Isoengelitin34.650.70RenshenP69MOL007180Vitamin-e32.290.70SharenP70MOL000953CLR37.870.68Yiyiren, shanyaoP71MOL000554Gallic acid-3-O-(6′-O-galloyl)-glucoside30.250.67Fuling, sharenP72MOL002311Glycyrol90.780.67GancaoP73MOL01145520-Hexadecanoylingenol32.700.65Renshen, fulingP74MOL004904Licopyranocoumarin80.360.65GancaoP75MOL0049591-Methoxyphaseollidin69.980.64GancaoP76MOL004071Hyndarin73.940.64GancaoP77MOL005360Malkangunin57.710.63Renshen, baizhuP78MOL004824(2S)-6-(2, 4-dihydroxyphenyl)-2-(2-hydroxypropan-2-yl)-4-methoxy-2, 3-dihydrofuro[3, 2-g]chromen-7-one60.250.63GancaoP79MOL005008Glycyrrhiza flavonol A41.280.60GancaoP80MOL005007Glyasperins M72.670.59GancaoP81MOL004492Chrysanthemaxanthin38.720.58Renshen, fulingP82MOL005017Phaseol78.770.58GancaoP83MOL005003Licoagrocarpin58.810.58GancaoP84MOL002773Beta-carotene37.180.58BaibiandouP85MOL0049743′-methoxyglabridin46.160.57GancaoP86MOL0049663′-hydroxy-4′-O-Methylglabridin43.710.57GancaoP87MOL004806Euchrenone30.290.57GancaoP88MOL005384Suchilactone57.520.56Renshen, baizhuP89MOL005344Ginsenoside rh236.320.56RenshenP90MOL006982Codeine45.480.56SharenP91MOL004827Semilicoisoflavone B48.780.55GancaoP92MOL004884Licoisoflavone B38.930.55GancaoP93MOL0049053, 22-Dihydroxy-11-oxo-delta(12)-oleanene-27-alpha-methoxycarbonyl-29-oic acid34.320.55GancaoP94MOL003648Inermine65.830.54RenshenP95MOL004810Glyasperin F75.840.54GancaoP96MOL001484Inermine75.180.54GancaoP97MOL004885Licoisoflavanone52.470.54GancaoP98MOL005461Doradexanthin38.160.54ShanyaoP99MOL0049141, 3-Dihydroxy-8, 9-dimethoxy-6-benzofurano [3, 2-c]chromenone62.900.53GancaoP100MOL004820Kanzonols W50.480.52GancaoP101MOL0049782-[(3R)-8, 8-Dimethyl-3, 4-dihydro-2h-pyrano [6, 5-f]chromen-3-yl]-5-methoxyphenol36.210.52GancaoP102MOL003851Isoramanone39.970.51GancaoP103MOL004912Glabrone52.510.50GancaoP104MOL005314Celabenzine101.880.49RenshenP105MOL005012Licoagroisoflavone57.280.49GancaoP106MOL004855Licoricone63.580.47GancaoP107MOL004908Glabridin53.250.47GancaoP108MOL004879Glycyrin52.610.47GancaoP109MOL009436Stemotinine38.690.46BaizhuP110MOL004857Gancaonin B48.790.45GancaoP111MOL004833Phaseolinisoflavan32.010.45GancaoP112MOL004808Glyasperin B65.220.44GancaoP113MOL004911Glabrene46.270.44GancaoP114MOL001002Ellagic acid43.060.43Fuling, sharenP115MOL0048493-(2, 4-Dihydroxyphenyl)-8-(1, 1-dimethylprop-2-enyl)-7-hydroxy-5-methoxy-coumarin59.620.43GancaoP116MOL0049131, 3-Dihydroxy-9-methoxy-6-benzofurano [3, 2-c]chromenone48.140.43GancaoP117MOL008118Coixenolide32.400.43YiyirenP118MOL004949Isolicoflavonol45.170.42GancaoP119MOL004883Licoisoflavone41.610.42GancaoP120MOL004814Isotrifoliol31.940.42GancaoP121MOL002372(6Z, 10E, 14E, 18E)-2, 6, 10, 15, 19, 23-Hexamethyltetracosa-2, 6, 10, 14, 18, 22-hexaene33.550.42YiyirenP122MOL0048633-(3, 4-Dihydroxyphenyl)-5, 7-dihydroxy-8-(3-methylbut-2-enyl)chromone66.370.41GancaoP123MOL0048662-(3, 4-Dihydroxyphenyl)-5, 7-dihydroxy-6-(3-methylbut-2-enyl)chromone44.150.41GancaoP124MOL0049896-Prenylated eriodictyol39.220.41GancaoP125MOL004935Sigmoidin-B34.880.41GancaoP126MOL0048645, 7-Dihydroxy-3-(4-methoxyphenyl)-8-(3-methylbut-2-enyl)chromone30.490.41GancaoP127MOL005890Pachypodol75.060.40FulingP128MOL0049938-Prenylated eriodictyol53.790.40GancaoP129MOL004856Gancaonin A51.080.40GancaoP130MOL004811Glyasperin C45.560.40GancaoP131MOL007213Nuciferine34.430.40LianziP132MOL012537Spinoside A41.750.40JiegengP133MOL008406Spinoside A39.970.40JiegengP134MOL002879Diop43.590.39RenshenP135MOL005000Gancaonin G60.440.39GancaoP136MOL005430Hancinone C59.050.39ShanyaoP137MOL0048388-(6-Hydroxy-2-benzofuranyl)-2, 2-dimethyl-5-chromenol58.440.38GancaoP138MOL006980Papaverine64.040.38SharenP139MOL000322Kadsurenone54.720.38ShanyaoP140MOL000310Denudatin B61.470.38ShanyaoP141MOL005020Dehydroglyasperins C53.820.37GancaoP142MOL003656Lupiwighteone51.640.37GancaoP143MOL004915Eurycarpin A43.280.37GancaoP144MOL009172Pronuciferin32.750.37LianziP145MOL005429Hancinol64.010.37ShanyaoP146MOL004882Licocoumarone33.210.36GancaoP147MOL003673Wighteone42.800.36GancaoP148MOL004907Glyzaglabrin61.070.35GancaoP149MOL004828Glepidotin A44.720.35GancaoP150MOL004815(E)-1-(2, 4-dihydroxyphenyl)-3-(2, 2-dimethylchromen-6-yl)prop-2-en-1-one39.620.35GancaoP151MOL005321Frutinone A65.900.34RenshenP152MOL004829Glepidotin B64.460.34GancaoP153MOL002565Medicarpin49.220.34GancaoP154MOL011072Quinicine75.440.33Fuling, baibiandouP155MOL004961Quercetin der.46.450.33GancaoP156MOL004980Inflacoumarin A39.710.33GancaoP157MOL004848Licochalcone G49.250.32GancaoP158MOL004945(2S)-7-hydroxy-2-(4-hydroxyphenyl)-8-(3-methylbut-2-enyl)chroman-4-one36.570.32GancaoP159MOL005356Girinimbin61.220.31RenshenP160MOL00002114-Acetyl-12-senecioyl-2E, 8E, 10E-atractylentriol60.310.31BaizhuP161MOL004910Glabranin52.900.31GancaoP162MOL000354Isorhamnetin49.600.31Gancao, baibiandouP163MOL004898(E)-3-[3, 4-dihydroxy-5-(3-methylbut-2-enyl)phenyl]-1-(2, 4-dihydroxyphenyl)prop-2-en-1-one46.270.31GancaoP164MOL00002214-Acetyl-12-senecioyl-2E, 8Z, 10E-atractylentriol63.370.30BaizhuP165MOL005016Odoratin49.950.30GancaoP166MOL002882[(2R)-2, 3-dihydroxypropyl] (Z)-octadec-9-enoate34.130.30YiyirenP167MOL000239Jaranol50.830.29GancaoP168MOL000497Licochalcone a40.790.29GancaoP169MOL007206Armepavine69.310.29LianziP170MOL0081212-Monoolein34.230.29YiyirenP171MOL009135Ellipticine30.820.28Fuling, sharenP172MOL000098Quercetin46.430.28Gancao, sharen, baibiandouP173MOL004576Taxifolin57.840.27RenshenP174MOL0049907, 2′, 4′-Trihydroxy－5-methoxy-3-arylcoumarin83.710.27GancaoP175MOL004860Licorice glycoside E32.890.27GancaoP176MOL005575Gentiacaulein72.820.27GancaoP177MOL001735Dinatin30.970.27GancaoP178MOL004580cis-Dihydroquercetin66.440.27JiegengP179MOL001736(-)-Taxifolin60.510.27ShanyaoP180MOL005267Elymoclavine72.870.27ShanyaoP181MOL0049917-Acetoxy-2-methylisoflavone38.920.26GancaoP182MOL011093Apohyoscine59.680.25RenshenP183MOL003617Isogosferol30.070.25GancaoP184MOL000006Luteolin36.160.25JiegengP185MOL0059962-O-methyl-3―O-β-D-glucopyranosyl platycogenate A45.150.25JiegengP186MOL006026Dimethyl 2-O-methyl-3-O-a-D-glucopyranosyl platycogenate A39.210.25JiegengP187MOL000422Kaempferol41.880.24Renshen, gancao, baibiandouP188MOL000417Calycosin47.750.24GancaoP189MOL005573Genkwanin37.130.24GancaoP190MOL000492(+)-Catechin54.830.24Sharen, baibiandouP191MOL001689Acacetin34.970.24JiegengP192MOL005463Methylcimicifugoside_qt31.690.24ShanyaoP193MOL007514Methyl icosa-11, 14-dienoate39.670.23SharenP194MOL003975Icosa-11, 14, 17-trienoic acid methyl ester44.810.23SharenP195MOL005308Aposiopolamine66.650.22RenshenP196MOL0000493β-acetoxyatractylone54.070.22BaizhuP197MOL00002012-Senecioyl-2E, 8 E, 10E-atractylentriol62.400.22BaizhuP198MOL0000728β-ethoxy atractylenolide III35.950.21BaizhuP199MOL010586Formononetin66.390.21BaizhuP200MOL000500Vestitol74.660.21GancaoP201MOL000392Formononetin69.670.21Gancao, baibiandouP202MOL004328Naringenin59.290.21Gancao, yiyiren, jiegengP203MOL004957HMO38.370.21GancaoP204MOL002419Demethylcoclaurine((R)-norcoclaurine)82.540.21LianziP205MOL005320Arachidonate45.570.20RenshenP206MOL005318Dianthramine40.450.20RenshenP207MOL0038967-Methoxy-2-methyl isoflavone42.560.20GancaoP208MOL004985Icos-5-enoic acid30.700.20GancaoP209MOL004996Gadelaidic acid30.700.20GancaoP210MOL000230Pinocembrin57.560.20GancaoP211MOL004841Licochalcone B76.760.19GancaoP212MOL004835Glypallichalcone61.600.19GancaoP213MOL001494Mandenol42.000.19YiyirenP214MOL004058Khell33.190.19ShanyaoP215MOL004941(2R)-7-hydroxy-2-(4-hydroxyphenyl)chroman-4-one71.120.18GancaoP216MOL001792DFV32.760.18GancaoP217MOL001559Piperlonguminine30.710.18Shanyao


### 3.2. Collection of BA and AC Disease Targets

4795 BA-related target genes were collected based on DisGeNET, TTD, GeneCards, PharmGKB, OMIM, NCBI, The Human Phenotype Ontology, and DrugBank databases. Duplicate targets were excavated and deleted, and 3388 BA disease action targets in total were collected. 1828 AC-related target genes were collected based on TTD, GeneCards, PharmGKB, OMIM, and NCBI databases. And 1640 AC disease action targets in total were collected by mining and deleting duplicate targets. The obtained target information was standardized for gene symbol and UniProt ID.


### 3.3. Acquirement of Targets of Active Compounds of SLBZP for Treating BA and AC

After searching the above-mentioned qualified potential active compounds of SLBZP in the TCMSP database, and removing the repeated targets, 281 targets of active compounds of SLBZP were obtained. The bioinformatics and evolutionary genomics platform was used to match the potential targets of drugs with disease targets, and a Venn diagram was drawn (Figure2). 149 common targets were obtained (Table 2).Figure 2
Targets matching among SLBZP, BA, and AC.Table 2
Characteristics of common targets.
No.TargetSymbolUniProt IDNo.TargetSymbolUniProt ID172 kDa type IV collagenaseMMP2P0825376Prostaglandin E2 receptor EP3 subtypePTGER3P431152Xanthine dehydrogenase/oxidaseXDHP4798977Urokinase-type plasminogen activatorPLAUP007493Heat shock protein beta-1HSPB1P0479278Phosphatidylinositol-3,4,5-trisphosphate 3-phosphatase and dual-specificity protein phosphatase PTENPTENP604844Nitric oxide synthase, inducibleNOS2P3522879Sodium-dependent serotonin transporterSLC6A4P316455Hepatocyte growth factor receptorMETP0858180Interferon regulatory factor 1IRF1P109146UDP-glucuronosyltransferase 1–1UGT1A1P2230981Arachidonate 5-lipoxygenaseALOX5P099177Protein kinase C beta typePRKCBP0577182Gap junction alpha-1 proteinGJA1P173028Collagen alpha-1(I) chainCOL1A1P0245283Claudin-4CLDN4O144939Baculoviral IAP repeat-containing protein 5BIRC5O1539284Dipeptidyl peptidase IVDPP4P2748710Apoptosis regulator Bcl-2BCL2P1041585Serum paraoxonase/arylesterase 1PON1P2716911Alpha-2A adrenergic receptorADRA2AP0891386Caspase-8CASP8Q1479012Cytochrome P450 1A1CYP1A1P0479887Peroxisome proliferator activated receptor gammaPPARGP37231135-Hydroxytryptamine receptor 3AHTR3AP4609888C-X-C motif chemokine 11CXCL11O1462514Mitogen-activated protein kinase 10MAPK10P5377989Interleukin-8CXCL8P1014515ProstaglandinE synthasePTGESO1468490E-selectinSELEP1658116C-reactive proteinCRPP0274191ThrombomodulinTHBDP0720417Glutathione S-transferase PGSTP1P0921192Glucocorticoid receptorNR3C1P0415018Aryl hydrocarbon receptorAHRP3586993Serine/threonine-protein kinase mTORMTORP4234519Nuclear factor erythroid 2-related factor 2NFE2L2Q1623694Mitogen-activated protein kinase 14MAPK14Q1653920Tumor necrosis factorTNFP0137595RAF proto-oncogene serine/threonine-protein kinaseRAF1P0404921Pro-epidermal growth factorEGFP0113396Cytosolic phospholipase A2PLA2G4AP4771222Interleukin-1 alphaIL1AP0158397MyeloperoxidaseMPOP0516423Canalicular multispecific organic anion transporter 1ABCC2Q9288798Alpha-1B adrenergic receptorADRA1BP3536824Caspase-1CASP1P2946699Inhibitor of nuclear factor kappa-B kinase subunit alphaCHUKO1511125OsteopontinSPP1P10451100Signal transducer and activator of transcription 3STAT3P4076326ThrombinF2P00734101AntileukoproteinaseSLPIP0397327Prostaglandin G/H synthase 2PTGS2P35354102Cathepsin DCTSDP0733928Catenin beta-1CTNNB1P35222103Sterol O-acyltransferase 1SOAT1P3561029G1/S-specific cyclin-D1CCND1P24385104AcetylcholinesteraseACHEP2230330Estrogen receptorESR1P03372105Induced myeloid leukemia cell differentiation protein Mcl-1MCL1Q0782031Vascular endothelial growth factor AVEGFAP15692106C-C motif chemokine 2CCL2P1350032Transforming growth factor beta-1TGFB1P01137107Interleukin-6IL6P0523133Myc proto-oncogene proteinMYCP01106108Caspase-3CASP3P4257434Cyclin-A2CCNA2P20248109Heat shock protein HSP 90-alphaHSP90AA1P0790035Glycogen synthase kinase-3 betaGSK3BP49841110Poly [ADP-ribose] polymerase 1PARP1P0987436Interstitial collagenaseMMP1P03956111Tumor necrosis factor ligand superfamily member 6FASLGP4802337Signal transducer and activator of transcription 1-alpha/betaSTAT1P42224112Maltase-glucoamylase, intestinalMGAMO4345138Peroxisome proliferator activated receptor deltaPPARDQ03181113Vascular endothelial growth factor receptor 2KDRP35968393-Hydroxy-3-methylglutaryl-coenzyme a reductaseHMGCRP04035114Fos-related antigen 2FOSL2P1540840Mineralocorticoid receptorNR3C2P08235115ATP-binding cassette sub-familyG member 2ABCG2Q9UNQ041Glutathione reductase, mitochondrialGSRP00390116Peroxisome proliferator-activated receptor alphaPPARAQ0786942Heme oxygenase 1HMOX1P09601117Cytochrome P450 1A2CYP1A2P0517743Stromelysin-1MMP3P08254118Insulin-like growth factor IIIGF2P0134444Pituitary adenylate cyclase-activating polypeptideADCYAP1P18509119Phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit, gamma isoformPIK3CGP4873645Glutathione S-transferase mu 1GSTM1P09488120NAD(P)H dehydrogenase [quinone] 1NQO1P1555946Interleukin-10IL10P22301121Interleukin-2IL2P6056847Mitogen-activated protein kinase 1MAPK1P28482122Receptor tyrosine-protein kinase erbB-3ERBB3P2186048C-X-C motif chemokine 2CXCL2P19875123Interferon gammaIFNGP0157949Epidermal growth factor receptorEGFRP00533124Proto-oncogene c-FosFOSP0110050Inhibitor of nuclear factor kappa-B kinase subunit betaIKBKBO1492012578 kDa glucose-regulated proteinHSPA5P1102151Superoxide dismutase [Cu-Zn]SOD1P00441126Intercellular adhesion molecule 1ICAM1P0536252Receptor tyrosine-protein kinase erbB-2ERBB2P04626127Caveolin-1CAV1Q0313553Interleukin-4IL4P05112128Bcl-2-like protein 1BCL2L1Q0781754Mitogen-activated protein kinase 8MAPK8P45983129Mitogen-activated protein kinase 3MAPK3P2736155Aldose reductaseAKR1B1P15121130Carbonic anhydrase IICA2P0091856Histamine H1 receptorHRH1P35367131Transcription factor p65RELAQ0420657Cell division protein kinase 2CDK2P24941132Hypoxia-inducible factor 1-alphaHIF1AQ1666558Progesterone receptorPGRP06401133Nitric-oxide synthase, endothelialNOS3P2947459Ornithine decarboxylaseODC1P11926134Mu-type opioid receptorOPRM1P3537260C-X-C motif chemokine 10CXCL10P02778135Plasminogen activator inhibitor 1SERPINE1P0512161Cellular tumor antigen p53TP53P04637136Vascular cell adhesion protein 1VCAM1P1932062Caspase-9CASP9P55211137RAC-alpha serine/threonine-protein kinaseAKT1P3174963Cyclin-dependent kinase inhibitor 1CDKN1AP38936138Prostaglandin G/H synthase 1PTGS1P2321964CatalaseCATP04040139Tissue factorF3P1372665NAD-dependent deacetylase sirtuin-1SIRT1Q96EB6140Nuclear receptor sub-family 1 group I member 2NR1I2O7546966Multidrug resistance-associated protein 1ABCC1P33527141Transcription factor AP-1JUNP0541267Interleukin-1 betaIL1BP01584142Androgen receptorARP1027568NF-kappa-B inhibitor alphaNFKBIAP25963143Apoptosis regulator BAXBAXQ0781269Insulin-like growth factor-binding protein 3IGFBP3P17936144Protein kinase C alpha typePRKCAP1725270Serum albuminALBP02768145CD40 ligandCD40LGP29965715-Hydroxytryptamine 2A receptorHTR2AP28223146Cytochrome P450 3A4CYP3A4P0868472Stromelysin-2MMP10P09238147Matrix metalloproteinase-9MMP9P1478073Estrogen receptor betaESR2Q92731148AdiponectinADIPOQQ1584874Cytochrome P450 1B1CYP1B1Q16678149Retinoic acid receptor RXR-betaRXRBP2870275Neuronal acetylcholine receptor protein, alpha-7 chainCHRNA7P36544


### 3.4. GO and KEGG Pathway Enrichment Analysis

GO enrichment analysis revealed 2933 biological functions with remarkable significance, including 2687 for biological processes (BP), 75 for cellular component (CC), and 171 for molecular function (MF). The results of GO enrichment analysis showed that the common targets of SLBZP in treating BA and AC mainly involved response to oxidative stress, response to molecule of bacterial origin, membrane region, membrane microdomain, signaling receptor activator activity, receptor ligand activity, and other biological functions (Figure3). 182 significant pathways were obtained by KEGG pathway enrichment analysis, mainly involving PI3K-Akt signaling pathway, proteoglycans in cancer, MAPK signaling pathway, IL-17 signaling pathway, TNF signaling pathway, apoptosis, Th17 cell differentiation, and other pathways related to inflammation, cancer, apoptosis, and immunity (Figure 4).Figure 3
GO enrichment analysis of common targets.Figure 4
KEGG pathway enrichment analysis of common targets.


### 3.5. Construction and Analysis of Active Compound-Key Target Network

The 149 common targets obtained above were screened by “cytoHubba”, a plug-in of Cytoscape software, and then the 20 key targets with the highest degree value were obtained (Figure5). These 20 key targets and their corresponding active compounds were imported into Cytoscape software for network construction and visualization (Figure 6). There were 131 active compounds related to key targets (Table 3). In the active compound-key target network, the degree of the network topology analysis carried out by “Network Analysis” reflects the connectivity of nodes that respectively represent active compounds and key targets. A higher degree value indicates more associations between nodes, which explains the significances of active compounds and key targets. The results of network topology analysis showed that the 5 active compounds most connected to the key targets were quercetin, luteolin, beta-carotene, kaempferol, and naringenin, and the top 6 key targets of connectivity were prostaglandin G/H synthase 2 (PTGS2), caspase-3 (CASP3), RAC-alpha serine/threonine-protein kinase (AKT1), transcription factor AP-1 (JUN) [, cellular tumor antigen p53 (TP53), and vascular endothelial growth factor A (VEGFA), which indicated that the above compounds and targets were critical and had important implications in SLBZP for treating BA and AC.Figure 5
PPI diagram of key targets.Figure 6
Diagram of active compound-key target network.Table 3
Active compounds related to key targets.
CodeMolecule IDMolecule nameOB (%)DLDegreeHerbsP172MOL000098Quercetin46.430.2816Gancao, sharen, baibiandouP184MOL000006Luteolin36.160.2511JiegengP84MOL002773Beta-carotene37.180.587BaibiandouP187MOL000422Kaempferol41.880.246Renshen, gancao, baibiandouP202MOL004328Naringenin59.290.215Gancao, yiyiren, jiegengP20MOL000546Diosgenin80.880.814ShanyaoP89MOL005344Ginsenoside rh236.320.564RenshenP171MOL009135Ellipticine30.820.283Fuling, sharenP191MOL001689Acacetin34.970.243JiegengP114MOL001002Ellagic acid43.060.433Fuling, sharenP42MOL000358Beta-sitosterol36.910.753Renshen, sharenP168MOL000497Licochalcone a40.790.293GancaoP201MOL000392Formononetin69.670.212Gancao, baibiandouP151MOL005321Frutinone A65.900.341RenshenP94MOL003648Inermine65.830.541RenshenP159MOL005356Girinimbin61.220.311RenshenP10MOL000787Fumarine59.260.831RenshenP88MOL005384Suchilactone57.520.561Renshen, baizhuP205MOL005320Arachidonate45.570.201RenshenP34MOL000449Stigmasterol43.830.761ShanyaoP206MOL005318Dianthramine40.450.201RenshenP68MOL004567Isoengelitin34.650.701RenshenP173MOL004576Taxifolin57.840.271RenshenP64MOL000493Campesterol37.580.711RenshenP164MOL00002214-Acetyl-12-senecioyl-2E,8Z,10E-atractylentriol63.370.301BaizhuP196MOL0000493β-acetoxyatractylone54.070.221BaizhuP198MOL0000728β-ethoxy atractylenolide III35.950.211BaizhuP199MOL010586Formononetin66.390.211BaizhuP53MOL009387Didehydrotuberostemonine51.910.741BaizhuP45MOL000296Hederagenin36.910.751FulingP59MOL009149Cheilanthifoline46.510.721FulingP154MOL011072Quinicine75.440.331Fuling, baibiandouP72MOL002311Glycyrol90.780.671GancaoP174MOL0049907,2′,4′-trihydroxy-5-methoxy-3-arylcoumarin83.710.271GancaoP74MOL004904Licopyranocoumarin80.360.651GancaoP56MOL004891Shinpterocarpin80.300.731GancaoP82MOL005017Phaseol78.770.581GancaoP211MOL004841Licochalcone B76.760.191GancaoP95MOL004810Glyasperin F75.840.541GancaoP96MOL001484Inermine75.180.541GancaoP200MOL000500Vestitol74.660.211GancaoP80MOL005007Glyasperins M72.670.591GancaoP215MOL004941(2R)-7-hydroxy-2-(4-hydroxyphenyl)chroman-4-one71.120.181GancaoP75MOL0049591-Methoxyphaseollidin69.980.641GancaoP122MOL0048633-(3,4-dihydroxyphenyl)-5,7-dihydroxy-8-(3-methylbut-2-enyl)chromone66.370.411GancaoP54MOL004903Liquiritin65.690.741GancaoP112MOL004808Glyasperin B65.220.441GancaoP152MOL004829Glepidotin B64.460.341GancaoP106MOL004855Licoricone63.580.471GancaoP212MOL004835Glypallichalcone61.600.191GancaoP148MOL004907Glyzaglabrin61.070.351GancaoP135MOL005000Gancaonin G60.440.391GancaoP78MOL004824(2S)-6-(2,4-dihydroxyphenyl)-2-(2-hydroxypropan-2-yl)-4-methoxy-2,3-dihydrofuro[3,2-g]chromen-7-one60.250.631GancaoP115MOL0048493-(2,4-dihydroxyphenyl)-8-(1,1-dimethylprop-2-enyl)-7-hydroxy-5-methoxy-coumarin59.620.431GancaoP83MOL005003Licoagrocarpin58.810.581GancaoP137MOL0048388-(6-Hydroxy-2-benzofuranyl)-2,2-dimethyl-5-chromenol58.440.381GancaoP105MOL005012Licoagroisoflavone57.280.491GancaoP3MOL005018Xambioona54.850.871GancaoP141MOL005020Dehydroglyasperins C53.820.371GancaoP128MOL0049938-Prenylated eriodictyol53.790.401GancaoP107MOL004908Glabridin53.250.471GancaoP161MOL004910Glabranin52.900.311GancaoP108MOL004879Glycyrin52.610.471GancaoP103MOL004912Glabrone52.510.501GancaoP97MOL004885Licoisoflavanone52.470.541GancaoP142MOL003656Lupiwighteone51.640.371GancaoP129MOL004856Gancaonin A51.080.401GancaoP167MOL000239Jaranol50.830.291GancaoP100MOL004820Kanzonols W50.480.521GancaoP30MOL005001Gancaonin H50.100.781GancaoP165MOL005016Odoratin49.950.301GancaoP162MOL000354Isorhamnetin49.600.311Gancao, baibiandouP157MOL004848Licochalcone G49.250.321GancaoP153MOL002565Medicarpin49.220.341GancaoP110MOL004857Gancaonin B48.790.451GancaoP91MOL004827Semilicoisoflavone B48.780.551GancaoP188MOL000417Calycosin47.750.241GancaoP155MOL004961Quercetin der.46.450.331GancaoP163MOL004898(E)-3-[3,4-dihydroxy-5-(3-methylbut-2-enyl)phenyl]-1-(2,4-dihydroxyphenyl)prop-2-en-1-one46.270.311GancaoP113MOL004911Glabrene46.270.441GancaoP85MOL0049743′-methoxyglabridin46.160.571GancaoP130MOL004811Glyasperin C45.560.401GancaoP118MOL004949Isolicoflavonol45.170.421GancaoP149MOL004828Glepidotin A44.720.351GancaoP8MOL004948Isoglycyrol44.700.841GancaoP123MOL0048662-(3,4-dihydroxyphenyl)-5,7-dihydroxy-6-(3-methylbut-2-enyl)chromone44.150.411GancaoP86MOL0049663′-hydroxy-4′-O-Methylglabridin43.710.571GancaoP143MOL004915Eurycarpin A43.280.371GancaoP207MOL0038967-Methoxy-2-methyl isoflavone42.560.201GancaoP119MOL004883Licoisoflavone41.610.421GancaoP79MOL005008Glycyrrhiza flavonol A41.280.601GancaoP1MOL004924(-)-Medicocarpin40.990.951GancaoP156MOL004980Inflacoumarin A39.710.331GancaoP150MOL004815(E)-1-(2,4-dihydroxyphenyl)-3-(2,2-dimethylchromen-6-yl)prop-2-en-1-one39.620.351GancaoP124MOL0049896-Prenylated eriodictyol39.220.411GancaoP92MOL004884Licoisoflavone B38.930.551GancaoP181MOL0049917-Acetoxy-2-methylisoflavone38.920.261GancaoP203MOL004957HMO38.370.211GancaoP158MOL004945(2S)-7-hydroxy-2-(4-hydroxyphenyl)-8-(3-methylbut-2-enyl)chroman-4-one36.570.321GancaoP101MOL0049782-[(3R)-8,8-dimethyl-3,4-dihydro-2h-pyrano [6,5-f]chromen-3-yl]-5-methoxyphenol36.210.521GancaoP125MOL004935Sigmoidin-B34.880.411GancaoP216MOL001792DFV32.760.181GancaoP2MOL004988Kanzonol F32.470.891GancaoP111MOL004833Phaseolinisoflavan32.010.451GancaoP120MOL004814Isotrifoliol31.940.421GancaoP60MOL004805(2S)-2-[4-hydroxy-3-(3-methylbut-2-enyl)phenyl]-8,8-dimethyl-2,3-dihydropyrano [2,3-f]chromen-4-one31.790.721GancaoP126MOL0048645,7-Dihydroxy-3-(4-methoxyphenyl)-8-(3-methylbut-2-enyl)chromone30.490.411GancaoP87MOL004806Euchrenone30.290.571GancaoP210MOL000230Pinocembrin57.560.201GancaoP189MOL005573Genkwanin37.130.241GancaoP176MOL005575Gentiacaulein72.820.271GancaoP147MOL003673Wighteone42.800.361GancaoP177MOL001735Hispidulin30.970.271GancaoP183MOL003617Isogosferol30.070.251GancaoP76MOL004071Tetrahydropalmatine73.940.641GancaoP169MOL007206Armepavine69.310.291LianziP144MOL009172Pronuciferine32.750.371LianziP131MOL007213Nuciferine34.430.401LianziP31MOL001323Sitosterol alpha143.280.781YiyirenP213MOL001494Mandenol42.000.191YiyirenP6MOL001474Sanguinarine37.810.861SharenP138MOL006980Papaverine64.040.381SharenP190MOL000492(+)-Catechin54.830.241Sharen, baibiandouP178MOL004580Cis-dihydroquercetin66.440.271JiegengP9MOL008752Dihydroverticillatine42.690.841JiegengP179MOL001736(-)-Taxifolin60.510.271ShanyaoP136MOL005430Hancinone C59.050.391ShanyaoP139MOL000322Kadsurenone54.720.381ShanyaoP33MOL005465AIDS18090745.330.771ShanyaoP180MOL005267Elymoclavine72.870.271ShanyaoP214MOL004058Deltoside33.190.191Shanyao


### 3.6. Molecular Docking Results

Based on the above analysis results, the 5 important active compounds (quercetin, luteolin, beta-carotene, kaempferol, and naringenin) and the key targets were docked by AutoDockVina software. The docking results are shown in Table4 and Figure 7. The smaller the binding free energy value, the lower the energy required for binding, which is more conducive to the binding of ligand and protein. Among them, the docking results of MMP9 with luteolin, quercetin, and kaempferol, ALB with luteolin, and PTGS2 with luteolin were the best, as shown in Figure 8. For example, luteolin formed conventional hydrogen bonds with MMP9 protein structure 6ESM amino acid residues A chain TYR245, LEU243, GLN227, LEU188, ALA189, formed π-σ interactions with amino acid residues A chain TYR248 and LEU188, formed π-π stacked interactions with amino acid residue A chain HIS226, and formed π-alkyl interactions with amino acid residues A chain VAL223 and LEU188. These forces reduced the binding energy and increased the affinity, which played an auxiliary role in the binding of compound ligand molecules to the residues of target protein structures.Table 4
Docking results of target proteins and active compounds.
Target proteinsPDB IDCompoundsBinding energy (kcal/mol)Target proteinsPDB IDCompoundsBinding energy (kcal/mol)IL61ALUQuercetin−7.2AKT11UNQQuercetin−6.2Luteolin−7.2Luteolin−6.3Beta-carotene−7.6Beta-carotene−6.9Kaempferol−6.8Kaempferol−6.0Naringenin−6.9Naringenin−7.0ALB6YG9Quercetin−9.8TP535MHCQuercetin−7.4Luteolin−10.1Luteolin−7.9Beta-carotene−8.2Beta-carotene−9.1Kaempferol−8.8Kaempferol−7.6Naringenin−8.2Naringenin−7.2VEGFA1MKKQuercetin−7.4TNF5UUIQuercetin−6.9Luteolin−7.8Luteolin−7.0Beta-carotene−7.6Beta-carotene−7.3Kaempferol−7.3Kaempferol−6.9Naringenin−7.5Naringenin−6.4MAPK34QTBQuercetin−9.3CASP32DKOQuercetin−7.0Luteolin−9.5Luteolin−6.9Beta-carotene−9.0Beta-carotene−6.2Kaempferol−9.3Kaempferol−6.7Naringenin−7.9Naringenin−6.5JUN6Y3VQuercetin−6.5PTGS25F19Quercetin−9.7Luteolin−6.5Luteolin−10.0Beta-carotene−7.3Beta-carotene−8.7Kaempferol−6.3Kaempferol−9.3Naringenin−6.5Naringenin−8.2STAT36NJSQuercetin−8.2MAPK82XRWQuercetin−8.2Luteolin−8.0Luteolin−8.6Beta-carotene−7.0Beta-carotene−9.7Kaempferol−7.9Kaempferol−8.6Naringenin−7.2Naringenin−6.4MMP96ESMQuercetin−10.7CXCL84XDXQuercetin−7.5Luteolin−10.9Luteolin−7.7Beta-carotene−8.8Beta-carotene−9.2Kaempferol−10.3Kaempferol−7.6Naringenin−8.7Naringenin−6.7EGFR5HG8Quercetin−8.3MAPK16SLGQuercetin−8.1Luteolin−8.6Luteolin−8.3Beta-carotene−9.2Beta-carotene−8.4Kaempferol−8.5Kaempferol−8.1Naringenin−7.9Naringenin−7.5EGF1JL9Quercetin−6.6MYC6G6KQuercetin−7.2Luteolin−6.8Luteolin−7.9Beta-carotene−6.8Beta-carotene−7.8Kaempferol−6.8Kaempferol−7.6Naringenin−5.9Naringenin−7.4IL1B5R8QQuercetin−7.1FOS1A02Quercetin−8.3Luteolin−7.8Luteolin−9.3Beta-carotene−7.8Beta-carotene−8.8Kaempferol−7.0Kaempferol−8.1Naringenin−6.9Naringenin−9.3Figure 7
Heat map of docking results between key targets and important active compounds.Figure 8
3D and 2D diagrams of molecular docking. (a) MMP9 (6ESM) and luteolin. (b) MMP9 (6ESM) and quercetin. (c) MMP9 (6ESM) and kaempferol. (d) ALB (6YG9) and luteolin. (e) PTGS2 (5F19) and luteolin.
(a)(b)(c)(d)(e)


### 3.7. Construction and Analysis of Key Target-miRNA Network

94 miRNAs were predicted from 6 key targets by the miRTarBase database. Cytoscape software was used to construct a key target-miRNA network (Figure9), among which hsa-miR-16-5p, hsa-miR-101-3p, hsa-miR-143-3p, hsa-miR-199a-5p, hsa-miR-30d-5p, hsa-miR-30c-5p, hsa-miR-30e-5p, hsa-miR-302d-3p, hsa-miR-203a-3p, hsa-miR-200b-3p, hsa-miR-125a-5p, hsa-miR-15a-5p, hsa-miR-504-5p, and hsa-miR-150-5p all targeted multiple key targets.Figure 9
Diagram of key target-miRNA network.


## 3.1. Acquirement of Active Compounds of SLBZP

Preliminarily, a total of 335 active compounds were acquired from the BATMAN-TCM database; a total of 443 active compounds were acquired from the ETCM database; a total of 1182 active compounds were acquired from the SymMap database; a total of 352 active compounds were acquired from the Traditional Chinese Medicine Database@TAIWAN database; and a total of 171 active compounds were acquired from the TCMSP database. At last, 217 eligible unique active compounds of SLBZP in total were retrieved from the TCMSP database under the conditions of OB ≥ 30% and DL ≥ 0.18, which are all shown in Table1.Table 1
Characteristics of eligible active compounds in SLBZP with OB and DL parameters.
CodeMolecule IDMolecule nameOB (%)DLHerbsP1MOL004924(-)-Medicocarpin40.990.95GancaoP2MOL004988Kanzonol F32.470.89GancaoP3MOL005018Xambioona54.850.87GancaoP4MOL005458Dioscoreside C_qt36.380.87ShanyaoP5MOL007536Stigmasta-5, 22-dien-3-beta-yl acetate46.440.86SharenP6MOL001474Sanguinarine37.810.86SharenP7MOL001973Sitosteryl acetate40.390.85SharenP8MOL004948Isoglycyrol44.70.84GancaoP9MOL008752Dihydroverticillatine42.690.84JiegengP10MOL000787Fumarine59.260.83RenshenP11MOL005357Gomisin B31.990.83RenshenP12MOL000300Dehydroeburicoic acid44.170.83FulingP13MOL000285(2R)-2-[(5R, 10S, 13R, 14R, 16R, 17R)-16-hydroxy-3-keto-4, 4, 10, 13, 14-pentamethyl-1, 2, 5, 6, 12, 15, 16, 17-octahydrocyclopenta[a]phenanthren-17-yl]-5-isopropyl-hex-5-enoic acid38.260.82FulingP14MOL000280(2R)-2-[(3S, 5R, 10S, 13R, 14R, 16R, 17R)-3, 16-dihydroxy-4, 4, 10, 13, 14-pentamethyl-2, 3, 5, 6, 12, 15, 16, 17-octahydro-1h-cyclopenta[a]phenanthren-17-yl]-5-isopropyl-hex-5-enoic acid31.070.82FulingP15MOL005317Deoxyharringtonine39.270.81RenshenP16MOL000283Ergosterol peroxide40.360.81FulingP17MOL0002873beta-hydroxy-24-methylene-8-lanostene-21-oic acid38.70.81FulingP18MOL0002767, 9(11)-Dehydropachymic acid35.110.81FulingP19MOL000289Pachymic acid33.630.81FulingP20MOL000546Diosgenin80.880.81ShanyaoP21MOL000275Trametenolic acid38.710.80FulingP22MOL005376Panaxadiol33.090.79RenshenP23MOL005401Ginsenoside Rg5_qt39.560.79RenshenP24MOL004917Glycyroside37.250.79GancaoP25MOL007535(5S, 8S, 9S, 10R, 13R, 14S, 17R)-17-[(1R, 4R)-4-ethyl-1, 5-dimethylhexyl]-10, 13-dimethyl-2, 4, 5, 7, 8, 9, 11, 12, 14, 15, 16, 17-dodecahydro-1h-cyclopenta[a]phenanthrene-3, 6-dione33.120.79SharenP26MOL005348Ginsenoside-Rh4_qt31.110.78RenshenP27MOL000033(3S, 8S, 9S, 10R, 13R, 14S, 17R)-10, 13-dimethyl-17-[(2R, 5S)-5-propan-2-yloctan-2-yl]-2, 3, 4, 7, 8, 9, 11, 12, 14, 15, 16, 17-dodecahydro-1h-cyclopenta[a]phenanthren-3-ol36.230.78BaizhuP28MOL009136Peraksine82.580.78FulingP29MOL000211Mairin55.380.78GancaoP30MOL005001Gancaonin H50.100.78GancaoP31MOL001323Sitosterol alpha143.280.78YiyirenP32MOL000279Cerevisterol37.960.77FulingP33MOL005465AIDS18090745.330.77ShanyaoP34MOL000449Stigmasterol43.830.76Renshen, yiyiren, sharen, baibiandou, shanyaoP35MOL000028α-Amyrin39.510.76BaizhuP36MOL000290Poricoic acid A30.610.76FulingP37MOL00175524-Ethylcholest-4-en-3-one36.080.76RenshenP38MOL004355Spinasterol42.980.76JiegengP39MOL004718α-Spinasterol42.980.76JiegengP40MOL005440Isofucosterol43.780.76ShanyaoP41MOL01062524-Methylenecholesterol43.540.76ShanyaoP42MOL000358Beta-sitosterol36.910.75Renshen, sharenP43MOL005399Alexandrin_qt36.910.75RenshenP44MOL001525Daucosterol36.910.75RenshenP45MOL000296Hederagenin36.910.75FulingP46MOL000292Poricoic acid C38.150.75FulingP47MOL000291Poricoic acid B30.520.75FulingP48MOL0063767-Dehydrosigmasterol37.420.75FulingP49MOL000359Sitosterol36.910.75Gancao, yiyirenP50MOL001771Poriferast-5-en-3beta-ol36.910.75SharenP51MOL013119Enhydrin40.560.74RenshenP52MOL000139Smitilbin37.600.74RenshenP53MOL009387Didehydrotuberostemonine51.910.74BaizhuP54MOL004903Liquiritin65.690.74GancaoP55MOL009154Tuberostemoenone53.900.73BaizhuP56MOL004891Shinpterocarpin80.300.73GancaoP57MOL009431Stemonine81.750.72BaizhuP58MOL000282Ergosta-7, 22e-dien-3beta-ol43.510.72FulingP59MOL009149Cheilanthifoline46.510.72FulingP60MOL004805(2S)-2-[4-hydroxy-3-(3-methylbut-2-enyl)phenyl]-8, 8-dimethyl-2, 3-dihydropyrano[2, 3-f]chromen-4-one31.790.72GancaoP61MOL00543524-Methylcholest-5-enyl-3belta-O-glucopyranoside_qt37.580.72ShanyaoP62MOL012254Campesterol37.580.71RenshenP63MOL005438Campesterol37.580.71Renshen, shanyaoP64MOL000493Campesterol37.580.71RenshenP65MOL00501318α-Hydroxyglycyrrhetic acid41.160.71GancaoP66MOL006070Robinin39.840.71JiegengP67MOL01104218Alpha-hydroglycyrrhetic acid38.930.71BaibiandouP68MOL004567Isoengelitin34.650.70RenshenP69MOL007180Vitamin-e32.290.70SharenP70MOL000953CLR37.870.68Yiyiren, shanyaoP71MOL000554Gallic acid-3-O-(6′-O-galloyl)-glucoside30.250.67Fuling, sharenP72MOL002311Glycyrol90.780.67GancaoP73MOL01145520-Hexadecanoylingenol32.700.65Renshen, fulingP74MOL004904Licopyranocoumarin80.360.65GancaoP75MOL0049591-Methoxyphaseollidin69.980.64GancaoP76MOL004071Hyndarin73.940.64GancaoP77MOL005360Malkangunin57.710.63Renshen, baizhuP78MOL004824(2S)-6-(2, 4-dihydroxyphenyl)-2-(2-hydroxypropan-2-yl)-4-methoxy-2, 3-dihydrofuro[3, 2-g]chromen-7-one60.250.63GancaoP79MOL005008Glycyrrhiza flavonol A41.280.60GancaoP80MOL005007Glyasperins M72.670.59GancaoP81MOL004492Chrysanthemaxanthin38.720.58Renshen, fulingP82MOL005017Phaseol78.770.58GancaoP83MOL005003Licoagrocarpin58.810.58GancaoP84MOL002773Beta-carotene37.180.58BaibiandouP85MOL0049743′-methoxyglabridin46.160.57GancaoP86MOL0049663′-hydroxy-4′-O-Methylglabridin43.710.57GancaoP87MOL004806Euchrenone30.290.57GancaoP88MOL005384Suchilactone57.520.56Renshen, baizhuP89MOL005344Ginsenoside rh236.320.56RenshenP90MOL006982Codeine45.480.56SharenP91MOL004827Semilicoisoflavone B48.780.55GancaoP92MOL004884Licoisoflavone B38.930.55GancaoP93MOL0049053, 22-Dihydroxy-11-oxo-delta(12)-oleanene-27-alpha-methoxycarbonyl-29-oic acid34.320.55GancaoP94MOL003648Inermine65.830.54RenshenP95MOL004810Glyasperin F75.840.54GancaoP96MOL001484Inermine75.180.54GancaoP97MOL004885Licoisoflavanone52.470.54GancaoP98MOL005461Doradexanthin38.160.54ShanyaoP99MOL0049141, 3-Dihydroxy-8, 9-dimethoxy-6-benzofurano [3, 2-c]chromenone62.900.53GancaoP100MOL004820Kanzonols W50.480.52GancaoP101MOL0049782-[(3R)-8, 8-Dimethyl-3, 4-dihydro-2h-pyrano [6, 5-f]chromen-3-yl]-5-methoxyphenol36.210.52GancaoP102MOL003851Isoramanone39.970.51GancaoP103MOL004912Glabrone52.510.50GancaoP104MOL005314Celabenzine101.880.49RenshenP105MOL005012Licoagroisoflavone57.280.49GancaoP106MOL004855Licoricone63.580.47GancaoP107MOL004908Glabridin53.250.47GancaoP108MOL004879Glycyrin52.610.47GancaoP109MOL009436Stemotinine38.690.46BaizhuP110MOL004857Gancaonin B48.790.45GancaoP111MOL004833Phaseolinisoflavan32.010.45GancaoP112MOL004808Glyasperin B65.220.44GancaoP113MOL004911Glabrene46.270.44GancaoP114MOL001002Ellagic acid43.060.43Fuling, sharenP115MOL0048493-(2, 4-Dihydroxyphenyl)-8-(1, 1-dimethylprop-2-enyl)-7-hydroxy-5-methoxy-coumarin59.620.43GancaoP116MOL0049131, 3-Dihydroxy-9-methoxy-6-benzofurano [3, 2-c]chromenone48.140.43GancaoP117MOL008118Coixenolide32.400.43YiyirenP118MOL004949Isolicoflavonol45.170.42GancaoP119MOL004883Licoisoflavone41.610.42GancaoP120MOL004814Isotrifoliol31.940.42GancaoP121MOL002372(6Z, 10E, 14E, 18E)-2, 6, 10, 15, 19, 23-Hexamethyltetracosa-2, 6, 10, 14, 18, 22-hexaene33.550.42YiyirenP122MOL0048633-(3, 4-Dihydroxyphenyl)-5, 7-dihydroxy-8-(3-methylbut-2-enyl)chromone66.370.41GancaoP123MOL0048662-(3, 4-Dihydroxyphenyl)-5, 7-dihydroxy-6-(3-methylbut-2-enyl)chromone44.150.41GancaoP124MOL0049896-Prenylated eriodictyol39.220.41GancaoP125MOL004935Sigmoidin-B34.880.41GancaoP126MOL0048645, 7-Dihydroxy-3-(4-methoxyphenyl)-8-(3-methylbut-2-enyl)chromone30.490.41GancaoP127MOL005890Pachypodol75.060.40FulingP128MOL0049938-Prenylated eriodictyol53.790.40GancaoP129MOL004856Gancaonin A51.080.40GancaoP130MOL004811Glyasperin C45.560.40GancaoP131MOL007213Nuciferine34.430.40LianziP132MOL012537Spinoside A41.750.40JiegengP133MOL008406Spinoside A39.970.40JiegengP134MOL002879Diop43.590.39RenshenP135MOL005000Gancaonin G60.440.39GancaoP136MOL005430Hancinone C59.050.39ShanyaoP137MOL0048388-(6-Hydroxy-2-benzofuranyl)-2, 2-dimethyl-5-chromenol58.440.38GancaoP138MOL006980Papaverine64.040.38SharenP139MOL000322Kadsurenone54.720.38ShanyaoP140MOL000310Denudatin B61.470.38ShanyaoP141MOL005020Dehydroglyasperins C53.820.37GancaoP142MOL003656Lupiwighteone51.640.37GancaoP143MOL004915Eurycarpin A43.280.37GancaoP144MOL009172Pronuciferin32.750.37LianziP145MOL005429Hancinol64.010.37ShanyaoP146MOL004882Licocoumarone33.210.36GancaoP147MOL003673Wighteone42.800.36GancaoP148MOL004907Glyzaglabrin61.070.35GancaoP149MOL004828Glepidotin A44.720.35GancaoP150MOL004815(E)-1-(2, 4-dihydroxyphenyl)-3-(2, 2-dimethylchromen-6-yl)prop-2-en-1-one39.620.35GancaoP151MOL005321Frutinone A65.900.34RenshenP152MOL004829Glepidotin B64.460.34GancaoP153MOL002565Medicarpin49.220.34GancaoP154MOL011072Quinicine75.440.33Fuling, baibiandouP155MOL004961Quercetin der.46.450.33GancaoP156MOL004980Inflacoumarin A39.710.33GancaoP157MOL004848Licochalcone G49.250.32GancaoP158MOL004945(2S)-7-hydroxy-2-(4-hydroxyphenyl)-8-(3-methylbut-2-enyl)chroman-4-one36.570.32GancaoP159MOL005356Girinimbin61.220.31RenshenP160MOL00002114-Acetyl-12-senecioyl-2E, 8E, 10E-atractylentriol60.310.31BaizhuP161MOL004910Glabranin52.900.31GancaoP162MOL000354Isorhamnetin49.600.31Gancao, baibiandouP163MOL004898(E)-3-[3, 4-dihydroxy-5-(3-methylbut-2-enyl)phenyl]-1-(2, 4-dihydroxyphenyl)prop-2-en-1-one46.270.31GancaoP164MOL00002214-Acetyl-12-senecioyl-2E, 8Z, 10E-atractylentriol63.370.30BaizhuP165MOL005016Odoratin49.950.30GancaoP166MOL002882[(2R)-2, 3-dihydroxypropyl] (Z)-octadec-9-enoate34.130.30YiyirenP167MOL000239Jaranol50.830.29GancaoP168MOL000497Licochalcone a40.790.29GancaoP169MOL007206Armepavine69.310.29LianziP170MOL0081212-Monoolein34.230.29YiyirenP171MOL009135Ellipticine30.820.28Fuling, sharenP172MOL000098Quercetin46.430.28Gancao, sharen, baibiandouP173MOL004576Taxifolin57.840.27RenshenP174MOL0049907, 2′, 4′-Trihydroxy－5-methoxy-3-arylcoumarin83.710.27GancaoP175MOL004860Licorice glycoside E32.890.27GancaoP176MOL005575Gentiacaulein72.820.27GancaoP177MOL001735Dinatin30.970.27GancaoP178MOL004580cis-Dihydroquercetin66.440.27JiegengP179MOL001736(-)-Taxifolin60.510.27ShanyaoP180MOL005267Elymoclavine72.870.27ShanyaoP181MOL0049917-Acetoxy-2-methylisoflavone38.920.26GancaoP182MOL011093Apohyoscine59.680.25RenshenP183MOL003617Isogosferol30.070.25GancaoP184MOL000006Luteolin36.160.25JiegengP185MOL0059962-O-methyl-3―O-β-D-glucopyranosyl platycogenate A45.150.25JiegengP186MOL006026Dimethyl 2-O-methyl-3-O-a-D-glucopyranosyl platycogenate A39.210.25JiegengP187MOL000422Kaempferol41.880.24Renshen, gancao, baibiandouP188MOL000417Calycosin47.750.24GancaoP189MOL005573Genkwanin37.130.24GancaoP190MOL000492(+)-Catechin54.830.24Sharen, baibiandouP191MOL001689Acacetin34.970.24JiegengP192MOL005463Methylcimicifugoside_qt31.690.24ShanyaoP193MOL007514Methyl icosa-11, 14-dienoate39.670.23SharenP194MOL003975Icosa-11, 14, 17-trienoic acid methyl ester44.810.23SharenP195MOL005308Aposiopolamine66.650.22RenshenP196MOL0000493β-acetoxyatractylone54.070.22BaizhuP197MOL00002012-Senecioyl-2E, 8 E, 10E-atractylentriol62.400.22BaizhuP198MOL0000728β-ethoxy atractylenolide III35.950.21BaizhuP199MOL010586Formononetin66.390.21BaizhuP200MOL000500Vestitol74.660.21GancaoP201MOL000392Formononetin69.670.21Gancao, baibiandouP202MOL004328Naringenin59.290.21Gancao, yiyiren, jiegengP203MOL004957HMO38.370.21GancaoP204MOL002419Demethylcoclaurine((R)-norcoclaurine)82.540.21LianziP205MOL005320Arachidonate45.570.20RenshenP206MOL005318Dianthramine40.450.20RenshenP207MOL0038967-Methoxy-2-methyl isoflavone42.560.20GancaoP208MOL004985Icos-5-enoic acid30.700.20GancaoP209MOL004996Gadelaidic acid30.700.20GancaoP210MOL000230Pinocembrin57.560.20GancaoP211MOL004841Licochalcone B76.760.19GancaoP212MOL004835Glypallichalcone61.600.19GancaoP213MOL001494Mandenol42.000.19YiyirenP214MOL004058Khell33.190.19ShanyaoP215MOL004941(2R)-7-hydroxy-2-(4-hydroxyphenyl)chroman-4-one71.120.18GancaoP216MOL001792DFV32.760.18GancaoP217MOL001559Piperlonguminine30.710.18Shanyao


## 3.2. Collection of BA and AC Disease Targets

4795 BA-related target genes were collected based on DisGeNET, TTD, GeneCards, PharmGKB, OMIM, NCBI, The Human Phenotype Ontology, and DrugBank databases. Duplicate targets were excavated and deleted, and 3388 BA disease action targets in total were collected. 1828 AC-related target genes were collected based on TTD, GeneCards, PharmGKB, OMIM, and NCBI databases. And 1640 AC disease action targets in total were collected by mining and deleting duplicate targets. The obtained target information was standardized for gene symbol and UniProt ID.


## 3.3. Acquirement of Targets of Active Compounds of SLBZP for Treating BA and AC

After searching the above-mentioned qualified potential active compounds of SLBZP in the TCMSP database, and removing the repeated targets, 281 targets of active compounds of SLBZP were obtained. The bioinformatics and evolutionary genomics platform was used to match the potential targets of drugs with disease targets, and a Venn diagram was drawn (Figure2). 149 common targets were obtained (Table 2).Figure 2
Targets matching among SLBZP, BA, and AC.Table 2
Characteristics of common targets.
No.TargetSymbolUniProt IDNo.TargetSymbolUniProt ID172 kDa type IV collagenaseMMP2P0825376Prostaglandin E2 receptor EP3 subtypePTGER3P431152Xanthine dehydrogenase/oxidaseXDHP4798977Urokinase-type plasminogen activatorPLAUP007493Heat shock protein beta-1HSPB1P0479278Phosphatidylinositol-3,4,5-trisphosphate 3-phosphatase and dual-specificity protein phosphatase PTENPTENP604844Nitric oxide synthase, inducibleNOS2P3522879Sodium-dependent serotonin transporterSLC6A4P316455Hepatocyte growth factor receptorMETP0858180Interferon regulatory factor 1IRF1P109146UDP-glucuronosyltransferase 1–1UGT1A1P2230981Arachidonate 5-lipoxygenaseALOX5P099177Protein kinase C beta typePRKCBP0577182Gap junction alpha-1 proteinGJA1P173028Collagen alpha-1(I) chainCOL1A1P0245283Claudin-4CLDN4O144939Baculoviral IAP repeat-containing protein 5BIRC5O1539284Dipeptidyl peptidase IVDPP4P2748710Apoptosis regulator Bcl-2BCL2P1041585Serum paraoxonase/arylesterase 1PON1P2716911Alpha-2A adrenergic receptorADRA2AP0891386Caspase-8CASP8Q1479012Cytochrome P450 1A1CYP1A1P0479887Peroxisome proliferator activated receptor gammaPPARGP37231135-Hydroxytryptamine receptor 3AHTR3AP4609888C-X-C motif chemokine 11CXCL11O1462514Mitogen-activated protein kinase 10MAPK10P5377989Interleukin-8CXCL8P1014515ProstaglandinE synthasePTGESO1468490E-selectinSELEP1658116C-reactive proteinCRPP0274191ThrombomodulinTHBDP0720417Glutathione S-transferase PGSTP1P0921192Glucocorticoid receptorNR3C1P0415018Aryl hydrocarbon receptorAHRP3586993Serine/threonine-protein kinase mTORMTORP4234519Nuclear factor erythroid 2-related factor 2NFE2L2Q1623694Mitogen-activated protein kinase 14MAPK14Q1653920Tumor necrosis factorTNFP0137595RAF proto-oncogene serine/threonine-protein kinaseRAF1P0404921Pro-epidermal growth factorEGFP0113396Cytosolic phospholipase A2PLA2G4AP4771222Interleukin-1 alphaIL1AP0158397MyeloperoxidaseMPOP0516423Canalicular multispecific organic anion transporter 1ABCC2Q9288798Alpha-1B adrenergic receptorADRA1BP3536824Caspase-1CASP1P2946699Inhibitor of nuclear factor kappa-B kinase subunit alphaCHUKO1511125OsteopontinSPP1P10451100Signal transducer and activator of transcription 3STAT3P4076326ThrombinF2P00734101AntileukoproteinaseSLPIP0397327Prostaglandin G/H synthase 2PTGS2P35354102Cathepsin DCTSDP0733928Catenin beta-1CTNNB1P35222103Sterol O-acyltransferase 1SOAT1P3561029G1/S-specific cyclin-D1CCND1P24385104AcetylcholinesteraseACHEP2230330Estrogen receptorESR1P03372105Induced myeloid leukemia cell differentiation protein Mcl-1MCL1Q0782031Vascular endothelial growth factor AVEGFAP15692106C-C motif chemokine 2CCL2P1350032Transforming growth factor beta-1TGFB1P01137107Interleukin-6IL6P0523133Myc proto-oncogene proteinMYCP01106108Caspase-3CASP3P4257434Cyclin-A2CCNA2P20248109Heat shock protein HSP 90-alphaHSP90AA1P0790035Glycogen synthase kinase-3 betaGSK3BP49841110Poly [ADP-ribose] polymerase 1PARP1P0987436Interstitial collagenaseMMP1P03956111Tumor necrosis factor ligand superfamily member 6FASLGP4802337Signal transducer and activator of transcription 1-alpha/betaSTAT1P42224112Maltase-glucoamylase, intestinalMGAMO4345138Peroxisome proliferator activated receptor deltaPPARDQ03181113Vascular endothelial growth factor receptor 2KDRP35968393-Hydroxy-3-methylglutaryl-coenzyme a reductaseHMGCRP04035114Fos-related antigen 2FOSL2P1540840Mineralocorticoid receptorNR3C2P08235115ATP-binding cassette sub-familyG member 2ABCG2Q9UNQ041Glutathione reductase, mitochondrialGSRP00390116Peroxisome proliferator-activated receptor alphaPPARAQ0786942Heme oxygenase 1HMOX1P09601117Cytochrome P450 1A2CYP1A2P0517743Stromelysin-1MMP3P08254118Insulin-like growth factor IIIGF2P0134444Pituitary adenylate cyclase-activating polypeptideADCYAP1P18509119Phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit, gamma isoformPIK3CGP4873645Glutathione S-transferase mu 1GSTM1P09488120NAD(P)H dehydrogenase [quinone] 1NQO1P1555946Interleukin-10IL10P22301121Interleukin-2IL2P6056847Mitogen-activated protein kinase 1MAPK1P28482122Receptor tyrosine-protein kinase erbB-3ERBB3P2186048C-X-C motif chemokine 2CXCL2P19875123Interferon gammaIFNGP0157949Epidermal growth factor receptorEGFRP00533124Proto-oncogene c-FosFOSP0110050Inhibitor of nuclear factor kappa-B kinase subunit betaIKBKBO1492012578 kDa glucose-regulated proteinHSPA5P1102151Superoxide dismutase [Cu-Zn]SOD1P00441126Intercellular adhesion molecule 1ICAM1P0536252Receptor tyrosine-protein kinase erbB-2ERBB2P04626127Caveolin-1CAV1Q0313553Interleukin-4IL4P05112128Bcl-2-like protein 1BCL2L1Q0781754Mitogen-activated protein kinase 8MAPK8P45983129Mitogen-activated protein kinase 3MAPK3P2736155Aldose reductaseAKR1B1P15121130Carbonic anhydrase IICA2P0091856Histamine H1 receptorHRH1P35367131Transcription factor p65RELAQ0420657Cell division protein kinase 2CDK2P24941132Hypoxia-inducible factor 1-alphaHIF1AQ1666558Progesterone receptorPGRP06401133Nitric-oxide synthase, endothelialNOS3P2947459Ornithine decarboxylaseODC1P11926134Mu-type opioid receptorOPRM1P3537260C-X-C motif chemokine 10CXCL10P02778135Plasminogen activator inhibitor 1SERPINE1P0512161Cellular tumor antigen p53TP53P04637136Vascular cell adhesion protein 1VCAM1P1932062Caspase-9CASP9P55211137RAC-alpha serine/threonine-protein kinaseAKT1P3174963Cyclin-dependent kinase inhibitor 1CDKN1AP38936138Prostaglandin G/H synthase 1PTGS1P2321964CatalaseCATP04040139Tissue factorF3P1372665NAD-dependent deacetylase sirtuin-1SIRT1Q96EB6140Nuclear receptor sub-family 1 group I member 2NR1I2O7546966Multidrug resistance-associated protein 1ABCC1P33527141Transcription factor AP-1JUNP0541267Interleukin-1 betaIL1BP01584142Androgen receptorARP1027568NF-kappa-B inhibitor alphaNFKBIAP25963143Apoptosis regulator BAXBAXQ0781269Insulin-like growth factor-binding protein 3IGFBP3P17936144Protein kinase C alpha typePRKCAP1725270Serum albuminALBP02768145CD40 ligandCD40LGP29965715-Hydroxytryptamine 2A receptorHTR2AP28223146Cytochrome P450 3A4CYP3A4P0868472Stromelysin-2MMP10P09238147Matrix metalloproteinase-9MMP9P1478073Estrogen receptor betaESR2Q92731148AdiponectinADIPOQQ1584874Cytochrome P450 1B1CYP1B1Q16678149Retinoic acid receptor RXR-betaRXRBP2870275Neuronal acetylcholine receptor protein, alpha-7 chainCHRNA7P36544


## 3.4. GO and KEGG Pathway Enrichment Analysis

GO enrichment analysis revealed 2933 biological functions with remarkable significance, including 2687 for biological processes (BP), 75 for cellular component (CC), and 171 for molecular function (MF). The results of GO enrichment analysis showed that the common targets of SLBZP in treating BA and AC mainly involved response to oxidative stress, response to molecule of bacterial origin, membrane region, membrane microdomain, signaling receptor activator activity, receptor ligand activity, and other biological functions (Figure3). 182 significant pathways were obtained by KEGG pathway enrichment analysis, mainly involving PI3K-Akt signaling pathway, proteoglycans in cancer, MAPK signaling pathway, IL-17 signaling pathway, TNF signaling pathway, apoptosis, Th17 cell differentiation, and other pathways related to inflammation, cancer, apoptosis, and immunity (Figure 4).Figure 3
GO enrichment analysis of common targets.Figure 4
KEGG pathway enrichment analysis of common targets.


## 3.5. Construction and Analysis of Active Compound-Key Target Network

The 149 common targets obtained above were screened by “cytoHubba”, a plug-in of Cytoscape software, and then the 20 key targets with the highest degree value were obtained (Figure5). These 20 key targets and their corresponding active compounds were imported into Cytoscape software for network construction and visualization (Figure 6). There were 131 active compounds related to key targets (Table 3). In the active compound-key target network, the degree of the network topology analysis carried out by “Network Analysis” reflects the connectivity of nodes that respectively represent active compounds and key targets. A higher degree value indicates more associations between nodes, which explains the significances of active compounds and key targets. The results of network topology analysis showed that the 5 active compounds most connected to the key targets were quercetin, luteolin, beta-carotene, kaempferol, and naringenin, and the top 6 key targets of connectivity were prostaglandin G/H synthase 2 (PTGS2), caspase-3 (CASP3), RAC-alpha serine/threonine-protein kinase (AKT1), transcription factor AP-1 (JUN) [, cellular tumor antigen p53 (TP53), and vascular endothelial growth factor A (VEGFA), which indicated that the above compounds and targets were critical and had important implications in SLBZP for treating BA and AC.Figure 5
PPI diagram of key targets.Figure 6
Diagram of active compound-key target network.Table 3
Active compounds related to key targets.
CodeMolecule IDMolecule nameOB (%)DLDegreeHerbsP172MOL000098Quercetin46.430.2816Gancao, sharen, baibiandouP184MOL000006Luteolin36.160.2511JiegengP84MOL002773Beta-carotene37.180.587BaibiandouP187MOL000422Kaempferol41.880.246Renshen, gancao, baibiandouP202MOL004328Naringenin59.290.215Gancao, yiyiren, jiegengP20MOL000546Diosgenin80.880.814ShanyaoP89MOL005344Ginsenoside rh236.320.564RenshenP171MOL009135Ellipticine30.820.283Fuling, sharenP191MOL001689Acacetin34.970.243JiegengP114MOL001002Ellagic acid43.060.433Fuling, sharenP42MOL000358Beta-sitosterol36.910.753Renshen, sharenP168MOL000497Licochalcone a40.790.293GancaoP201MOL000392Formononetin69.670.212Gancao, baibiandouP151MOL005321Frutinone A65.900.341RenshenP94MOL003648Inermine65.830.541RenshenP159MOL005356Girinimbin61.220.311RenshenP10MOL000787Fumarine59.260.831RenshenP88MOL005384Suchilactone57.520.561Renshen, baizhuP205MOL005320Arachidonate45.570.201RenshenP34MOL000449Stigmasterol43.830.761ShanyaoP206MOL005318Dianthramine40.450.201RenshenP68MOL004567Isoengelitin34.650.701RenshenP173MOL004576Taxifolin57.840.271RenshenP64MOL000493Campesterol37.580.711RenshenP164MOL00002214-Acetyl-12-senecioyl-2E,8Z,10E-atractylentriol63.370.301BaizhuP196MOL0000493β-acetoxyatractylone54.070.221BaizhuP198MOL0000728β-ethoxy atractylenolide III35.950.211BaizhuP199MOL010586Formononetin66.390.211BaizhuP53MOL009387Didehydrotuberostemonine51.910.741BaizhuP45MOL000296Hederagenin36.910.751FulingP59MOL009149Cheilanthifoline46.510.721FulingP154MOL011072Quinicine75.440.331Fuling, baibiandouP72MOL002311Glycyrol90.780.671GancaoP174MOL0049907,2′,4′-trihydroxy-5-methoxy-3-arylcoumarin83.710.271GancaoP74MOL004904Licopyranocoumarin80.360.651GancaoP56MOL004891Shinpterocarpin80.300.731GancaoP82MOL005017Phaseol78.770.581GancaoP211MOL004841Licochalcone B76.760.191GancaoP95MOL004810Glyasperin F75.840.541GancaoP96MOL001484Inermine75.180.541GancaoP200MOL000500Vestitol74.660.211GancaoP80MOL005007Glyasperins M72.670.591GancaoP215MOL004941(2R)-7-hydroxy-2-(4-hydroxyphenyl)chroman-4-one71.120.181GancaoP75MOL0049591-Methoxyphaseollidin69.980.641GancaoP122MOL0048633-(3,4-dihydroxyphenyl)-5,7-dihydroxy-8-(3-methylbut-2-enyl)chromone66.370.411GancaoP54MOL004903Liquiritin65.690.741GancaoP112MOL004808Glyasperin B65.220.441GancaoP152MOL004829Glepidotin B64.460.341GancaoP106MOL004855Licoricone63.580.471GancaoP212MOL004835Glypallichalcone61.600.191GancaoP148MOL004907Glyzaglabrin61.070.351GancaoP135MOL005000Gancaonin G60.440.391GancaoP78MOL004824(2S)-6-(2,4-dihydroxyphenyl)-2-(2-hydroxypropan-2-yl)-4-methoxy-2,3-dihydrofuro[3,2-g]chromen-7-one60.250.631GancaoP115MOL0048493-(2,4-dihydroxyphenyl)-8-(1,1-dimethylprop-2-enyl)-7-hydroxy-5-methoxy-coumarin59.620.431GancaoP83MOL005003Licoagrocarpin58.810.581GancaoP137MOL0048388-(6-Hydroxy-2-benzofuranyl)-2,2-dimethyl-5-chromenol58.440.381GancaoP105MOL005012Licoagroisoflavone57.280.491GancaoP3MOL005018Xambioona54.850.871GancaoP141MOL005020Dehydroglyasperins C53.820.371GancaoP128MOL0049938-Prenylated eriodictyol53.790.401GancaoP107MOL004908Glabridin53.250.471GancaoP161MOL004910Glabranin52.900.311GancaoP108MOL004879Glycyrin52.610.471GancaoP103MOL004912Glabrone52.510.501GancaoP97MOL004885Licoisoflavanone52.470.541GancaoP142MOL003656Lupiwighteone51.640.371GancaoP129MOL004856Gancaonin A51.080.401GancaoP167MOL000239Jaranol50.830.291GancaoP100MOL004820Kanzonols W50.480.521GancaoP30MOL005001Gancaonin H50.100.781GancaoP165MOL005016Odoratin49.950.301GancaoP162MOL000354Isorhamnetin49.600.311Gancao, baibiandouP157MOL004848Licochalcone G49.250.321GancaoP153MOL002565Medicarpin49.220.341GancaoP110MOL004857Gancaonin B48.790.451GancaoP91MOL004827Semilicoisoflavone B48.780.551GancaoP188MOL000417Calycosin47.750.241GancaoP155MOL004961Quercetin der.46.450.331GancaoP163MOL004898(E)-3-[3,4-dihydroxy-5-(3-methylbut-2-enyl)phenyl]-1-(2,4-dihydroxyphenyl)prop-2-en-1-one46.270.311GancaoP113MOL004911Glabrene46.270.441GancaoP85MOL0049743′-methoxyglabridin46.160.571GancaoP130MOL004811Glyasperin C45.560.401GancaoP118MOL004949Isolicoflavonol45.170.421GancaoP149MOL004828Glepidotin A44.720.351GancaoP8MOL004948Isoglycyrol44.700.841GancaoP123MOL0048662-(3,4-dihydroxyphenyl)-5,7-dihydroxy-6-(3-methylbut-2-enyl)chromone44.150.411GancaoP86MOL0049663′-hydroxy-4′-O-Methylglabridin43.710.571GancaoP143MOL004915Eurycarpin A43.280.371GancaoP207MOL0038967-Methoxy-2-methyl isoflavone42.560.201GancaoP119MOL004883Licoisoflavone41.610.421GancaoP79MOL005008Glycyrrhiza flavonol A41.280.601GancaoP1MOL004924(-)-Medicocarpin40.990.951GancaoP156MOL004980Inflacoumarin A39.710.331GancaoP150MOL004815(E)-1-(2,4-dihydroxyphenyl)-3-(2,2-dimethylchromen-6-yl)prop-2-en-1-one39.620.351GancaoP124MOL0049896-Prenylated eriodictyol39.220.411GancaoP92MOL004884Licoisoflavone B38.930.551GancaoP181MOL0049917-Acetoxy-2-methylisoflavone38.920.261GancaoP203MOL004957HMO38.370.211GancaoP158MOL004945(2S)-7-hydroxy-2-(4-hydroxyphenyl)-8-(3-methylbut-2-enyl)chroman-4-one36.570.321GancaoP101MOL0049782-[(3R)-8,8-dimethyl-3,4-dihydro-2h-pyrano [6,5-f]chromen-3-yl]-5-methoxyphenol36.210.521GancaoP125MOL004935Sigmoidin-B34.880.411GancaoP216MOL001792DFV32.760.181GancaoP2MOL004988Kanzonol F32.470.891GancaoP111MOL004833Phaseolinisoflavan32.010.451GancaoP120MOL004814Isotrifoliol31.940.421GancaoP60MOL004805(2S)-2-[4-hydroxy-3-(3-methylbut-2-enyl)phenyl]-8,8-dimethyl-2,3-dihydropyrano [2,3-f]chromen-4-one31.790.721GancaoP126MOL0048645,7-Dihydroxy-3-(4-methoxyphenyl)-8-(3-methylbut-2-enyl)chromone30.490.411GancaoP87MOL004806Euchrenone30.290.571GancaoP210MOL000230Pinocembrin57.560.201GancaoP189MOL005573Genkwanin37.130.241GancaoP176MOL005575Gentiacaulein72.820.271GancaoP147MOL003673Wighteone42.800.361GancaoP177MOL001735Hispidulin30.970.271GancaoP183MOL003617Isogosferol30.070.251GancaoP76MOL004071Tetrahydropalmatine73.940.641GancaoP169MOL007206Armepavine69.310.291LianziP144MOL009172Pronuciferine32.750.371LianziP131MOL007213Nuciferine34.430.401LianziP31MOL001323Sitosterol alpha143.280.781YiyirenP213MOL001494Mandenol42.000.191YiyirenP6MOL001474Sanguinarine37.810.861SharenP138MOL006980Papaverine64.040.381SharenP190MOL000492(+)-Catechin54.830.241Sharen, baibiandouP178MOL004580Cis-dihydroquercetin66.440.271JiegengP9MOL008752Dihydroverticillatine42.690.841JiegengP179MOL001736(-)-Taxifolin60.510.271ShanyaoP136MOL005430Hancinone C59.050.391ShanyaoP139MOL000322Kadsurenone54.720.381ShanyaoP33MOL005465AIDS18090745.330.771ShanyaoP180MOL005267Elymoclavine72.870.271ShanyaoP214MOL004058Deltoside33.190.191Shanyao


## 3.6. Molecular Docking Results

Based on the above analysis results, the 5 important active compounds (quercetin, luteolin, beta-carotene, kaempferol, and naringenin) and the key targets were docked by AutoDockVina software. The docking results are shown in Table4 and Figure 7. The smaller the binding free energy value, the lower the energy required for binding, which is more conducive to the binding of ligand and protein. Among them, the docking results of MMP9 with luteolin, quercetin, and kaempferol, ALB with luteolin, and PTGS2 with luteolin were the best, as shown in Figure 8. For example, luteolin formed conventional hydrogen bonds with MMP9 protein structure 6ESM amino acid residues A chain TYR245, LEU243, GLN227, LEU188, ALA189, formed π-σ interactions with amino acid residues A chain TYR248 and LEU188, formed π-π stacked interactions with amino acid residue A chain HIS226, and formed π-alkyl interactions with amino acid residues A chain VAL223 and LEU188. These forces reduced the binding energy and increased the affinity, which played an auxiliary role in the binding of compound ligand molecules to the residues of target protein structures.Table 4
Docking results of target proteins and active compounds.
Target proteinsPDB IDCompoundsBinding energy (kcal/mol)Target proteinsPDB IDCompoundsBinding energy (kcal/mol)IL61ALUQuercetin−7.2AKT11UNQQuercetin−6.2Luteolin−7.2Luteolin−6.3Beta-carotene−7.6Beta-carotene−6.9Kaempferol−6.8Kaempferol−6.0Naringenin−6.9Naringenin−7.0ALB6YG9Quercetin−9.8TP535MHCQuercetin−7.4Luteolin−10.1Luteolin−7.9Beta-carotene−8.2Beta-carotene−9.1Kaempferol−8.8Kaempferol−7.6Naringenin−8.2Naringenin−7.2VEGFA1MKKQuercetin−7.4TNF5UUIQuercetin−6.9Luteolin−7.8Luteolin−7.0Beta-carotene−7.6Beta-carotene−7.3Kaempferol−7.3Kaempferol−6.9Naringenin−7.5Naringenin−6.4MAPK34QTBQuercetin−9.3CASP32DKOQuercetin−7.0Luteolin−9.5Luteolin−6.9Beta-carotene−9.0Beta-carotene−6.2Kaempferol−9.3Kaempferol−6.7Naringenin−7.9Naringenin−6.5JUN6Y3VQuercetin−6.5PTGS25F19Quercetin−9.7Luteolin−6.5Luteolin−10.0Beta-carotene−7.3Beta-carotene−8.7Kaempferol−6.3Kaempferol−9.3Naringenin−6.5Naringenin−8.2STAT36NJSQuercetin−8.2MAPK82XRWQuercetin−8.2Luteolin−8.0Luteolin−8.6Beta-carotene−7.0Beta-carotene−9.7Kaempferol−7.9Kaempferol−8.6Naringenin−7.2Naringenin−6.4MMP96ESMQuercetin−10.7CXCL84XDXQuercetin−7.5Luteolin−10.9Luteolin−7.7Beta-carotene−8.8Beta-carotene−9.2Kaempferol−10.3Kaempferol−7.6Naringenin−8.7Naringenin−6.7EGFR5HG8Quercetin−8.3MAPK16SLGQuercetin−8.1Luteolin−8.6Luteolin−8.3Beta-carotene−9.2Beta-carotene−8.4Kaempferol−8.5Kaempferol−8.1Naringenin−7.9Naringenin−7.5EGF1JL9Quercetin−6.6MYC6G6KQuercetin−7.2Luteolin−6.8Luteolin−7.9Beta-carotene−6.8Beta-carotene−7.8Kaempferol−6.8Kaempferol−7.6Naringenin−5.9Naringenin−7.4IL1B5R8QQuercetin−7.1FOS1A02Quercetin−8.3Luteolin−7.8Luteolin−9.3Beta-carotene−7.8Beta-carotene−8.8Kaempferol−7.0Kaempferol−8.1Naringenin−6.9Naringenin−9.3Figure 7
Heat map of docking results between key targets and important active compounds.Figure 8
3D and 2D diagrams of molecular docking. (a) MMP9 (6ESM) and luteolin. (b) MMP9 (6ESM) and quercetin. (c) MMP9 (6ESM) and kaempferol. (d) ALB (6YG9) and luteolin. (e) PTGS2 (5F19) and luteolin.
(a)(b)(c)(d)(e)


## 3.7. Construction and Analysis of Key Target-miRNA Network

94 miRNAs were predicted from 6 key targets by the miRTarBase database. Cytoscape software was used to construct a key target-miRNA network (Figure9), among which hsa-miR-16-5p, hsa-miR-101-3p, hsa-miR-143-3p, hsa-miR-199a-5p, hsa-miR-30d-5p, hsa-miR-30c-5p, hsa-miR-30e-5p, hsa-miR-302d-3p, hsa-miR-203a-3p, hsa-miR-200b-3p, hsa-miR-125a-5p, hsa-miR-15a-5p, hsa-miR-504-5p, and hsa-miR-150-5p all targeted multiple key targets.Figure 9
Diagram of key target-miRNA network.


## 4. Discussion

The theory of traditional Chinese medicine believes that the spleen is the foundation of acquired life and that the spleen is not harmonious and causes all kinds of diseases. Therefore, it has always paid attention to regulating the spleen to protect the five internal organs. The pathogenesis of spleen deficiency is involved in the occurrence and development of BA and AC. SLBZP, one of the classic Earth-cultivating and gold-generating prescriptions, can not only treat BA and AC with simultaneous treatment of different diseases but also protect the spleen to prevent and promote recovery. This study aimed to explore the action mechanism of SLBZP in treating BA and AC with simultaneous treatment of different diseases by using network pharmacology and molecular docking, so as to provide references for more in-depth experimental research and wider clinical applications.GO annotation results showed that the biological functions involved in common targets were mainly response to oxidative stress, response to molecule of bacterial origin, membrane region, membrane microdomain, signaling receptor activator activity, receptor ligand activity, and so on. In addition, the main enrichment pathways of common targets were PI3K-Akt signaling pathway, proteoglycans in cancer, MAPK signaling pathway, IL-17 signaling pathway, TNF signaling pathway, apoptosis, Th17 cell differentiation, and other pathways related to inflammation, cancer, apoptosis, and immunity. Studies pointed out that, during the onset of asthma, both PI3K-Akt signaling pathway and MAPK signaling pathway were active [35, 36]. Many targets of the PI3K pathway play critical roles in the expression and activation of inflammatory mediators, inflammatory cell recruitment, immune cell function, airway remodeling, and corticosteroid insensitivity in chronic inflammatory airway disease [37]. There were evidences that selective PI3K inhibitors could reduce inflammation and some characteristics of diseases such as abnormal proliferation of airway smooth muscle cells (ASMC) in experimental animal models, which strongly supported that PI3K/Akt inhibitors might be a useful new therapy for asthma [37, 38]. In recent years, many studies confirmed that inhibiting PI3K−Akt signaling pathway and MAPK signaling pathway could effectively inhibit allergic airway inflammation, ASMC proliferation and migration, and phenotypic switching, so as to alleviate airway remodeling and airway hyperresponsiveness (AHR) in asthma [39–42]. Additionally, upregulation of dual-specificity phosphatase-1 (DUSP1), a negative regulator in the MAPK signaling pathway, to healthy levels and downregulation of inflammatory MAPKs at the gene and protein levels could reduce the prevalence of childhood asthma [43]. Proteoglycans enhanced deposition in the airway walls of asthmatics playing a role in airway remodeling, and the difference of deposition in the airway smooth muscle layer of moderate and severe asthmatic patients might affect the functional behavior of airway smooth muscle [44, 45]. IL-17A in the IL-17 signaling pathway was positively correlated with neutrophil accumulation, mucus secretion, macrophage mobilization, and smooth muscle reactivity in various experimental airway models, as well as with disease severity, suggesting that specifically targeting IL-17A had the potential of clinical utility in patients with moderate and severe asthma and high reversibility [46]. Moreover, the reduction of skin inflammation and airway inflammation in the IL-17-induced mouse asthma model was related to the reduction of IL-17-mediated mRNA stability [47]. In TLR ligand-mediated allergic airway inflammation, TLR ligand induced TNF to send signals through airway epithelial cells to promote the development of Th2 in lymph nodes, and TNF was also indispensable in the allergen stimulation stage of neutrophilic and eosinophilic airway inflammation and AHR [48]. Activated TNF-TNFR2 signal transduction could inhibit the differentiation of Th2 and Th17 cells to alleviate allergic airway inflammation [49]. Bronchial cell apoptosis could be observed in some airway biopsies from asthmatic patients, especially those with serious diseases, possibly resulting in airway damage, and dysregulation of leukocyte, eosinophil, and neutrophil apoptosis could lead to asthmatic airway inflammation and was related to the pathogenesis of asthma [50]. Th17 cells, a potent and unique subset that modulated primary bronchial epithelial cell function, were related to the development and pathophysiology of asthma [51, 52]. A study found that asthma-associated IL4R variants promoted the transformation of regulatory T cells into TH17-like cells, thereby exacerbating airway inflammation [53]. It should be noted that because there have been relatively few studies related to allergic colitis all the time, there is almost no relevant research report on the relationship between the above signaling pathways and allergic colitis. However, it is worth mentioning that if further research is carried out on this basis in the future, it will be very innovative and instructive for clarifying the pathogenesis of allergic colitis and developing new drugs that can effectively target the disease. The above showed that SLBZP treated BA and AC with simultaneous treatment of different diseases by multiple functions and pathways, suggesting that further research in the future could be based on these biological functions and pathways, which had guiding significances.The active compound-key target network of this study showed that the five active compounds of quercetin, luteolin, beta-carotene, kaempferol, and naringenin, and the 6 key targets of PTGS2, CASP3, AKT1, JUN, TP53, and VEGFA were particularly important. Moreover, the results of molecular docking also verified that these five active compounds had good binding characteristics with their corresponding important key targets, indicating that they played vital roles in SLBZP for treating BA and AC with simultaneous treatment of different diseases and had critical potential research values.Studies suggested that quercetin was known for its antioxidant activity in free radicals scavenging and antiallergic properties [54]. It is characterized by stimulating the immune system and antiviral activity, inhibiting histamine release, reducing proinflammatory cytokines, and producing leukotrienes [55]. It was reported to improve Th1/Th2 balance, inhibit the formation of antigen-specific IgE antibodies, and also be effective in inhibiting enzymes such as lipoxygenase, eosinophils, and peroxidase and inflammatory mediators [56]. All the mentioned mechanisms contribute to the anti-inflammatory and immunomodulatory properties of quercetin, which can be effectively used to treat advanced bronchial asthma, allergic rhinitis, and restrictive allergic reactions caused by peanuts [55]. Luteolin, having anti-inflammatory, antiallergic, and immune-enhancing functions, can reduce airway inflammation and allergies in asthma and has antiallergic effects in mouse models of allergic asthma and rhinitis, which has shown therapeutic effects in treating inflammatory diseases, allergies, bronchial asthma, and systemic damage caused by free radicals [57–59]. It was reported to block the activation of MAPK and NF-κB signaling pathways to protect ARPE-19 cells from the proliferation of IL-6, IL-8, sICAM-1, and MCP-1 stimulated by IL-1β, thereby alleviating the inflammatory response [60]. Kaempferol, having antioxidant, anti-inflammatory, anticancer, and antidiabetic effects, could effectively improve allergic and inflammatory airway diseases by interfering with NF-κB signal transduction, which may help alleviate the inflammatory response associated with Cox2 expression [61–63]. Naringenin, having immunomodulatory, anticancer, antimutation, anti-inflammatory, antioxidant, antiproliferative, antiarthritis, and anticarcinogenic effects, can be used for treating osteoporosis, cancer, cardiovascular disease, and rheumatoid arthritis, which exhibits lipid-lowering and insulin-like properties, can inhibit allergen-induced airway inflammation and airway responsiveness, and inhibit NF-κB activity in a mouse model of asthma [64–66]. The above results indicate that SLBZP can fully exert its therapeutic effect by the synergy of multiple compounds, multiple targets, and multiple pathways and provide more new clues for the development of traditional Chinese medicine monomers to treat BA and AC. In addition, the effects of beta-carotene in treating BA and AC are currently seldom studied and reported, which can be used as a direction for in-depth research in the future.PTGS2, as the most critical target in the network, is one of the key factors of cell response to inflammation and has long been considered to play a key role in the pathogenesis of respiratory inflammation, including asthma [67, 68]. In addition to its anti-inflammatory effect, it can also exert anti-inflammatory/bronchial protection functions in the airway and can be expressed quickly and powerfully in response to various proinflammatory cytokines and mediators [68]. Caspase-3 is necessary for the development of various tissues, playing an important role in neurogenesis, synaptic activity, neuron growth cone guidance, and glial development. It was reported to mediate many nonapoptotic functions in cells and cell death in the process of apoptosis, participate in T and B cell homeostasis in a way that did not depend on apoptosis, and protect compressed organs from cell death [69]. AKT1 ablation promoted the polarization of macrophage M1, which could affect the severity of inflammatory diseases, such as inflammatory bowel disease, and was related to the regulation of innate immunity and inflammation [70]. JUN, the activation of which is caused by the imbalance of pulmonary oxidation and antioxidation in asthma, is an important therapeutic target for allergic airway inflammation and a key transcription factor for the anti-inflammatory activity of dexamethasone and may be an important molecular mechanism of steroids in asthma and other chronic inflammatory lung diseases [71, 72]. As an important mediator of oncogenic β-catenin signaling in the intestine, JUN is not only involved in inflammatory response and tumorigenesis but is also related to the inflammatory response in mice with LPS-induced macrophages and DSS-induced colitis [73, 74]. TP53, as a tumor suppressor protein, can produce anti-inflammatory reactions in the lungs and has a potential therapeutic effect in pneumonia, whose dysfunction is associated with acute lung injury, acute respiratory distress syndrome, chronic obstructive pulmonary disease, pulmonary fibrosis, bronchial asthma, pulmonary hypertension, pneumonia and tuberculosis, and so on [75]. It often mutates in human cancers. After the mutations, it prolongs the activation of NF-κB and promotes chronic inflammation and inflammation-related colorectal cancer, which is also related to the occurrence and development of inflammatory bowel disease [76–78]. VEGFA plays a fundamental role in the physiological and pathophysiological forms of angiogenesis. During airway growth, the balance regulation of angiogenic growth factor and vascular inhibitory protein enables the lung to obtain rich blood supply [79]. However, during chronic inflammation, VEGF stimulates angiogenesis and edema and induces Th2 and eosinophilic inflammation, mucous metaplasia, subepithelial fibrosis, myocyte proliferation, and dendritic cell activation, which is a sign of asthma exacerbation and can be used as a target for treating lung diseases and inflammatory bowel diseases [79–82]. The above studies indicate that these six key targets deserve attention in the study of the molecular mechanism of SLBZP for treating BA and AC with simultaneous treatment of different diseases and can be used as potential research objects.The key target-miRNA network shows that hsa-miR-16-5p, hsa-miR-101-3p, hsa-miR-143-3p, hsa-miR-199a-5p, hsa-miR-30d-5p, hsa-miR-30c-5p, hsa-miR-30e-5p, hsa-miR-302d-3p, hsa-miR-203a-3p, hsa-miR-200b-3p, hsa-miR-125a-5p, hsa-miR-15a-5p, hsa-miR-504-5p, and hsa-miR-150-5p all target and regulate multiple key targets, which may have important upstream regulatory effects and are of great significance for the occurrence, development, and treatment of BA and AC. Studies suggested that baseline airway secretion signatures of hsa-miR-302d-3p and hsa-miR-612 were detected during rhinovirus (RV) infection that was the most common cause of asthma exacerbation and the most important early risk factor for asthma development after childhood in children, which was helpful to develop novel strategies for treating and monitoring respiratory conditions in all age groups [83]. The low tissue level of hsa-miR-200b-3p is related to the cytopathic inflammation caused by human cytomegalovirus infection [84]. Hsa-miR-15a-5p may play an important role in reducing retinal leukopenia by inhibiting inflammatory cell signals, which can be used as a potential target for the inhibition of inflammatory mediators in diabetic retinopathy [85]. In addition to these miRNAs that could target and regulate multiple key targets, hsa-miR-146a-5p, as one of the predicted miRNAs, was upregulated in asthmatic patients to inhibit the expression level of PDE7A, which might be involved in mediating the pathogenesis of asthma [86]. Upregulation of Hsa_circ_0005519 could inhibit the expression of has-let-7a-5p in CD4 T cells of asthmatic patients and promote the production of IL-13 and IL-6, thereby exacerbating asthma [87]. Combining hsa-miR-155-5p and has-miR-532-5p could predict changes in asthma budesonide (ICS) treatment response over time [88]. In allergic settings, the expressions of hsa-miR-139-5p and hsa-miR-542-3p significantly decreased, resulting in increasing expression of pro-inflammatory and antiviral response genes, which might be important during asthma exacerbations [89]. Hsa-miR-19b-3p decreased in the plasma of BA patients, and the ROC curve showed that it could be used as a biomarker for the diagnosis of BA [90]. Hsa-miR-20a-5p, one of the dysregulated miRNAs in asthmatic patients, targeted and inhibited the expression of HDAC4, suppressed the expressions of TNF-α, IL-1β, and IFN-γ, and promoted the production of IL-10, thereby reducing allergic inflammation [91]. Downregulation of hsa-miR-145-5p that increased airway smooth muscle cell proliferation was a risk factor for an early decline (ED) pattern of lung function growth in asthmatic children with chronic obstructive pulmonary disease (COPD) [92]. Once again, it is particularly noted that, at present, there are basically no relevant research reports on these predicted miRNAs related to AC, and there are also very few research reports on these predicted miRNAs related to BA, which means that this study not only provides new insights for in-depth understanding of the pathogenesis of BA and AC and formulating corresponding new treatment strategies but also provides a practical basis for future validation studies. In general, there are few reports on the above-mentioned miRNAs that have great research potentials.


## 5. Conclusions

In conclusion, network pharmacology and molecular docking technology demonstrated that SLBZP in treating BA and AC with simultaneous treatment of different diseases was a complex process involving multiple compounds, multiple targets, and multiple pathways. It may involve important active compounds and key targets represented by quercetin, luteolin, beta-carotene, kaempferol, naringenin, PTGS2, CASP3, AKT1, JUN, TP53, and VEGFA, may be related to inflammation, cancer, apoptosis, and immune-related pathways, and may involve the targeted regulation of multiple upstream miRNAs. These can provide references for future clinical and experimental studies.



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*Source: 4687788-2022-05-09.xml*