Study on the Potential Mechanism of Traditional Marine Chinese Medicine Beishashen in Preventing Lung Injury Induced by Radiotherapy and Chemotherapy

. Lung injury after chemoradiotherapy is a common adverse reaction that is caused by radiation and chemotherapy drugs. Beishashen is a traditional marine Chinese medicine with multiple pharmacological properties. Te previous research reported that it has a potential prevention efect on injuries caused by radiation and chemotherapy drugs, but the specifc mechanism is unknown. Consequently, this study is based on bioinformatics and other methods, focusing on exploring the potential targets and molecular mechanisms of Beishashen in preventing lung injury after radiotherapy and chemotherapy. Tis study used network pharmacology methods to identify the bioactive compounds of Beishashen in order to better understand its therapeutic radiation-induced lung injury (RILI) and chemotherapy-induced lung injury (CILI) molecular mechanisms. We found that the role of Beishashen in preventing RILI and CILI involved several main pathways, especially the PI3K-AKT signaling pathway. PTGS2, PIK3CG, and RXRA were considered as key targets. Molecular docking and molecular dynamics showed good binding between the active ingredients and key targets, especially between alloisoimperatorin and RXRA. Tis study revealed the role of Beishashen in some potential regulating signal pathways in the treatment and prevention of RILI and CILI. Quercetin and alloisoimperatorin were the main active ingredients with low toxicity and can efectively bind with key targets. Tese fndings provided important insights into the potential use of Beishashen for RILI and CILI.


Introduction
Malignant tumors remain one of the leading causes of death worldwide, and tumor prevention and treatment have increasingly become a major technological issue in the feld of life and health and the healthcare industry [1].Radiation therapy is an efective tool used in the treatment of tumors, but it can cause normal tissue damage, especially lung injury after radiotherapy for breast cancer, lung cancer, or other malignant tumors of the chest [2].In addition, many patients undergo synchronous chemotherapy to enhance the antitumor efcacy of radiation, which can lead to the exacerbation of lung injury, known as chemotherapy-induced lung injury (CILI) [3,4].With an incidence rate of 10% to 40%, radiation-induced lung injury (RILI) is the most frequent and harmful side efect of radiotherapy for chest tumors.In severe cases, it can cause respiratory failure and endanger life, making it a key limiting factor for radiation dose [5,6].Te clinical manifestations of CILI generally include fever, cough, and difculty breathing, and both cytotoxic and noncytotoxic drugs can cause lung injury [7].Chemotherapy drugs are an important cause of druginduced lung injury, accounting for about 10% of patients receiving chemotherapy [8].Currently, the clinical treatment of both RILI and CILI mainly involves symptomatic treatment.However, pure antibacterial therapy is inefective for RILI and CILI, and a large amount of glucocorticoids can alleviate early acute infammatory reactions, but their efcacy in late-stage radiation-induced pulmonary fbrosis is not ideal, and they are accompanied by adverse reactions such as secondary infection, immunosuppression, and an increased risk of tumor metastasis or recurrence [9].Cytotoxic drugs such as amifostine, interferon, ambroxol, and methotrexate also have certain clinical efects, but the many toxic side efects inherent in the drugs greatly limit their application, and their use can exacerbate or lead to exacerbation of lung injury [10][11][12][13].Terefore, the exploration and discovery of drugs that have the efect of preventing and treating radiation and chemotherapy damage and their transformation into clinical applications has become a challenging issue that urgently needs to be addressed in the current feld of cancer clinical practice.
Coasta-Glehnia-Root (Beishashen or Shashen) is the dry root of the Umbelliferae plant Glehnia littoralis Fr.Schmidt ex Miq.It benefts the stomach, nourishes the yin, and clears the lungs (this is a treatment method in traditional Chinese medicine that nourishes the negative fuid in the lungs, making it sufcient and achieving the goal of treating diseases), and stimulates fuid production.It is commonly used to treat lung heat and dry cough, cough and phlegm, stomach yin defciency, heat disease, fuid injury, dry throat and thirst, etc [14].It is mainly produced in coastal provinces of China, including Shandong, Liaoning, and Jiangsu provinces, as well as on beaches or beaches in South Korea, Japan, and the northwest of the United States.It is a traditional marine Chinese medicine in China, and current modern pharmacological research has found that Beishashen and its extracts have antioxidant, antitumor, antiinfammatory, antibacterial, and neuroprotective efects [15][16][17][18][19].In recent years, some scholars have found through data mining that Beishashen has potential therapeutic value in traditional Chinese medicine interventions for RILI, but their mechanisms of action in preventing and treating RILI and CILI have not been thoroughly studied [20].In addition, the clinical efcacy of traditional Chinese medicine in treating lung injury caused by radiotherapy and chemotherapy has been improved through syndrome diferentiation intervention, and traditional Chinese medicine has a potential utilization mechanism for the prevention and treatment of RILI and CILI [21,22].
Terefore, our present study aims to explore the possible main active ingredients, key targets, and potential molecular mechanisms of traditional marine Chinese medicine Beishashen in the prevention and treatment of RILI and CILI.Meanwhile, we investigate the animal experimental studies and clinical application of Beishashen and related prescriptions for CILI or RILI.Te fndings of the present study would provide a theoretical reference and scientifc basis for the screening of efective ingredients in Beishashen, the prevention and treatment of RILI and CILI, and the modernization of marine medicine research.Figure 1 displays the study's procedure.

Materials and Methods
Tis study will analyze the possible main active ingredients, key targets, and potential molecular mechanisms of Beishashen in preventing and treating RILI and CILI through network pharmacology, molecular docking, molecular dynamics, prediction of the pharmaceutical properties of active ingredients, quality analysis, and other methods and means.

Chemical Composition and Active Ingredients of
Beishashen.Te traditional Chinese medicine systems pharmacology database TCMSP was used to collect the active ingredients and targets of Beishashen, and to examine the retrieved ingredients' active ingredients.Oral bioavailability (OB) > 30% and drug similarity (DL) > 0.18 were the screening requirements [23].After obtaining the active ingredients and corresponding targets of Beishashen, the target proteins were merged, deweighted, and then standardized using the UniPort protein database.
All active ingredients' chemical compositions of Beishashen are included in the TCMSP database.Te TCMSP database also ofers downloads of the appropriate chemical structures of the active substances that were obtained.In addition, a MOL2-format molecular structure fle of the active ingredient was obtained for further use in molecular docking and molecular dynamics.

Acquisition of RILI and CILI Targets.
Te associated targets for RILI and CILI were searched using the GeneCards database and the Online Mendelian Inheritance in Man (OMIM) database.For instance, the relevant targets downloaded in databases were combined and then deduplicated to acquire the RILI relevant targets after "radiationinduced lung injury" was entered as a keyword in two databases.Only targets with relevant scores ≥10 were included in the GeneCards database, while all targets in the OMIM database were included.Following the use of R software, a Venn diagram of the Beishashen and RILI targets is drawn.Finally, the core targets of RILI were achieved.We also obtained CILI-related targets using the same procedure.

Construction of Protein-Protein Interaction.
On the strings platform, the protein-protein interaction (PPI) network was built.Te "Multiple Proteins" option was frst chosen, after which the name lists for the primary targets were added, and "Homo sapiens" was added to the organism query.Te PPI network diagram was eventually obtained after default values for other parameters were inserted.

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Journal of Food Biochemistry

Construction of Active Ingredient-Disease-Target
Network.We created a visual network graph of the active ingredient-disease-target network using Cytoscape 3.7.2software.Firstly, we input the obtained drugs, ingredients, diseases, and common targets into Cytoscape 3.7.2software.Subsequently, the network analyzer in the Cytoscape plugin was used to determine the "network nodes" topological properties.
Tese features included varying degrees of correlation, namely, degree, medium, and proximity.Besides, the fve highestassigned core targets were obtained based on degree values.Finally, the ingredient-disease-target network diagram was also generated during the aforementioned operation process.

KEGG Pathway Analysis and GO Functional Analysis.
Understanding the main targets' functions and important pathways of Beishashen prevention and treatment for RILI and CILI was made easier by Gene Ontology (GO) function analysis and Kyoto Gene and Genome Encyclopedia (KEGG) pathway analysis.We input core targets into the DAVID platform to obtain GO functional examination of cellular components, biological processes, and molecular functions.Subsequently, the top 10 items with the highest counts for each item were utilized to create a histogram with a p value using R software; it was statistically signifcant when p < 0.05.Similarly, we also obtained the outcomes of the DAVID platform's KEGG pathway analysis and drew a bar chart with the top 21 items with the highest count and p value; it was statistically signifcant when p < 0.05.

Molecular Docking.
Te molecular docking mechanism between the top fve major targets in the degree ranking of the PPI network and the major fve ingredients was verifed using AutoDock software.Te three-dimensional structures of the fve target proteins were sourced from the RCSB PDB protein structure database, while the MOL2 fle format of the active ingredients' structures were sourced from the TCMSP platform.In addition, the docking active site was located using the AutoDock plugin AutoGrid, and molecular docking was conducted to determine afnity.Te lower the binding energy, the better the binding afnity between the target and the active ingredient was when we employed binding energy ≤5.0 kJ•mol −1 as a key molecule with better binding afnity to key targets [24].Low-free binding energy docking data were visualized using the PyMOL program.

Molecular Dynamics.
Te molecular dynamics (MD) simulations were carried out by GROMACS 2020.3 software.Te simulation box size was optimized with the distance between each atom of the protein and the box greater than 1.0 nm.Ten, fll the box with water molecules based on a density of 1.To make the simulation system electrically neutral, the water molecules were replaced with Cl − and Na + ions.Following the steepest descent method, energy optimization of 5.0 × 10 4 steps was performed to minimize the energy consumption of the entire system and, fnally, to reduce the unreasonable contact or atom overlap in the entire system.After energy minimization, frst-phase equilibration was performed with the NVT ensemble at 300 K for 100 ps to stabilize the temperature of the system.Second-phase equilibration was simulated with the NPT ensemble at 1 bar and 100 ps.Te primary objective of the simulation is to optimize the interaction between the target protein and the solvent and ions so that the simulation system is fully preequilibrated.All MD simulations were performed for 100 ns under an isothermal and isostatic ensemble with a temperature of 300 K and a pressure of 1 atmosphere.Te temperature and pressure were controlled by the V-rescale and Parrinello-Rahman methods, respectively, and the temperature and pressure coupling constants were 0.1 and 0.5 ps, respectively.Lennard-Jones function was used to calculate the Van der Waals force, and the nonbond truncation distance was set to 1.4 nm.Te bond length of all atoms was constrained by the LINCS algorithm.
Te long-range electrostatic interaction was calculated by the particle Mesh-Ewald method with the Fourier spacing 0.16 nm.

ADMET Profling. Drug research and development heavily depend on understanding chemical absorption, distribution, metabolism, excretion, and toxicity (ADMET).
ADMETanalysis helps to achieve possibly the best balance of properties required for drug discovery, which is both efective and safe [25].In this regard, SWISS ADME and admetSAR were used to determine the physicochemical properties of the predicted ingredients.Tey were also used to predict the toxicity of potential active ingredients of Beishashen, such as acute oral toxicity, Ames mutagenesis, carcinogenic, hepatotoxicity, and nephrotoxicity.
2.9.Application of Beishashen and Related Prescriptions in RILI and CILI.In order to further understand the application of Beishashen and its prescriptions in RILI and CILI, we searched the relevant databases at home and abroad for their research reports on CILI and RILI in the past fve years, and their research results were briefy introduced and summarized.Terefore, we used a combination of free words and subject words to search PubMed, Web of Science, China National Knowledge Infrastructure (CNKI), China Science and Technology Journal Database (VIP), and Wanfang databases from January 1, 2018, to October 1st, 2023.English search terms included "Beishashen," "Shashen," "Radiation," "Lung injury," "Chemotherapy."Te research content is limited to clinical trials, clinical observational studies, clinical retrospective studies, and animal trials, but does not include reviews, pharmacokinetic studies, mechanism of action studies, meta-analysis, case reports, clinical guidelines, in vitro studies, etc. Te language of the literature is limited to English or Chinese.

Active Ingredients and Chemical Structure of Beishashen.
Eight of the functional ingredients and 184 associated targets of Beishashen were acquired from TCMSP in accordance with the screening criteria previously established.Moreover, the predicted targets' name was transformed into gene names through the UniPort database.In addition, we also collected the chemical compositions of the eight main active ingredients of Beishashen through the TCMSP database, and their MOL2 fles were also downloaded from the TCMSP database.Table 1 displays all of the facts in detail.

Te Targets of RILI and CILI.
GeneCards and the OMIM database were used to fnd the associated targets of RILI and CILI.While 452 associated targets were obtained from the OMIM database, 1927 related targets (relevance score 10) of RILI were obtained from the GeneCards database.2297 RILI-related targets were discovered after the merging of two datasets and deduplication.Te search and screening of CILI-related targets were also carried out according to the same process, with a total of 2264-related targets found from two databases.Besides, a Venn diagram was generated by using R software to identify the intersection of Beishashen active ingredient targets and the targets we obtained from RILI and CILI.We identifed 129 core targets that were shared between the Beishashen active ingredients and RILI/ CILI-related targets through this analysis.Figure 2 displays the outcomes.

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Journal of Food Biochemistry     promoter, positive regulation of translation, positive regulation of gene expression, negative regulation of the apoptotic process, DNA-templated, and positive regulation of cell promotion.Tey were distributed in the cyclosol, cycloplast, nucleus, plasma membrane, and nucleoplast.Tey also had molecular functions such as protein binding, identifying protein binding, enzyme binding, protein normalization activity, transcription factor activity, and sequence-specifc DNA binding.
Similar to the above analysis, key targets were subjected to KEGG pathway enrichment analysis using the DAVID platform and R software, and the top 25 with the highest count value were chosen for bar chart display.Te result was shown in Figure 6.Te main signal pathways involved include the PI3K-AKT signaling pathway, the AGE-RAGE signaling pathway in diabetic applications, the MAPK signaling pathway, the TNF signaling pathway, and the IL-17 signaling pathway.Te PI3K-AKT signaling pathway can be seen in Figure 7.

Molecular Docking Results
. We used molecular docking to compare the fve Beishashen active ingredients (alloissoimperatorin, quercetin, cnidilin, stigmasterol, and betasitosterol) and fve core targets (RXRA, PIK3CG, SCN5A, PTGS1, and PTGS2), respectively.Te outcomes of molecular docking demonstrated that each of the fve ingredients had strong interactions with fve target proteins and a high degree of matching, and all of the binding energy scores fell below −6.0 kJ•mol − .To determine the binding mode of the compound and target, we used Pymol2.1 software to view the compounds created by docking them with the targets.Te amino acid residues that the molecule mixes with the target pocket are also easily visible, depending on the binding mode.Among them, alloisoimperatorin had the highest binding energy with RXRA and PIK3CG, and both were less than −9.0 kJ•mol −1 .In addition, the results also showed that the amino acid residues of alloisoimperatorin interacting with the active site of RXRA included ASN-306, ALA-327, ARG-316, etc.In addition, alloisoimperatorin has a signifcant degree of hydrophobicity and is capable of forming a potent hydrophobic contact with the RXRA (TRP-305, LEU-436, LEU309, VAL-342, ILE-345, ILE324, PHE313, PHE-346, CYS432, VAL-349, ILE-268, LEU326, ALA-271, and ALA-272) active site amino acids, which is crucial for stabilizing small molecules in protein cavities.Table 2 displays the outcomes.

Molecular Dynamics.
Te outcomes of molecular docking showed that the RXRA and alloisoimperatorin complex had the highest binding energy value in its entirety to investigate the interaction mode between the key protein RXRA and the core ingredient alloisoimperatorin at a molecular level.Based on the outcomes of the molecular docking, we ran molecular dynamics simulations.Te root mean square deviation (RMSD) curve of the protein represented the fuctuation of RXRA conformation at diferent times.From Figure 8(a), it is apparent that during the initial stage of the molecular dynamics simulation (0-100 ns), the ligand RMSD value changed slightly while the protein RMSD value made a slight change.However, after 80 ns, the protein-ligand simulation system achieved equilibrium between the protein and ligand, and the protein RMSD value was greater than the ligand RMSD value, indicating that the protein fuctuations were greater than the ligand fuctuations.Te solvent accessible surface areas (SASA) were calculated by considering the interaction between van der Waals force and solvent molecules.Te solvent accessible surface area of the protein decreases with an increase in protein compactness, so the change of SASA can predict the change in protein structure (Figure 8(b)).As seen in Figure 8(b), the protein-ligand complex's SASA values showed an upward trend during the simulation of proteinligand recombination, indicating a decline in protein compactness.In addition, the radius of gyration (Rg) in Figure 8(c) also indicated that the Rg value and SASA value of the protein showed the same trend throughout the composite molecular dynamics simulation process, further signaling alterations to the protein-ligand system's structure.
We calculated the binding energy of all protein-ligand complexes in the equilibrium phase using the gmx_mmpbsa script to explain the interaction energy between the ligand and receptor.Te entire binding energy was split into four separate components for the MMPBSA application: electrostatic interaction, van der Waals interaction, polar solvation, and nonpolar solvation interaction.Note.A is the 3D structure of complex, B is the 2D binding mode of complex, C is the 3D binding mode of complex.
Journal of Food Biochemistry 9  number of molecular structures and properties to guide the selection of drugs that are most likely to be useful to patients.ADME analysis was conducted using the SWISS ADME database and the admetSAR database, and the results showed that all ingredients had satisfactory pharmacokinetic properties (Table 4).Te ADMET spectrum analysis results that some main active ingredients had slight toxic side efects in their pharmacokinetic properties, while others also had liver or kidney toxicity.Quercetin had the least toxic side efects, and there was no liver or kidney toxicity, while cnidilin had more toxic side efects.Te ADME characteristics of key active ingredients in diferent models, such as BBB permeant, P-gp substrate, CYP1A2 inhibitor, CYP2C19 inhibitor, and CYP2D6 inhibitor, had shown positive results in some potential ingredients, indicating their ability to serve as candidate drugs.Some compounds, such as quercetin and alloisoimperatorin, can be used as good candidate drugs for preclinical trials.

Te Application of Beishashen and Its Related Prescriptions in RILI and CILI.
According to our search conditions and requirements, a total of 10 articles about the application of the Beishashen prescription in clinical and animal experiments were included in the past 5 years, including 1 English paper and 9 Chinese papers.Tere is no research report on the use of Beishashen alone in recent 5 years.Beishashen is mostly studied in RILI and CILI in the form of traditional Chinese medicine prescriptions.In addition, there are 5 reports on the application of "Beishashen Maidong Decoction" as a research object in radiation pneumonia and lung injury; two articles reported the application of "Beishashen Jiegeng Decoction" as a research object in acute radiation-induced lung injury and radiation pneumonitis.Te detailed results are shown in Table 5.

Discussion
Te purpose of this work was to examine the probable mechanisms of Beishashen in the prevention and treatment of RILI and CILI using bioinformatics.Firstly, this work used the database to identify relevant targets for RILI and CILI after screening the active components and associated targets of Beishashen.Te intersection targets were obtained through a Venn diagram.Te team next built the active ingredient-disease-target network, performing enrichment analysis on the key targets.Based on the degree value, we identifed fve main active ingredients and fve main action targets.Tese fve active ingredients can successfully attach to those fve important targets, with quercetin and RXRA having the highest binding energies.Finally, the study predicted the pharmacokinetic characteristics of these fve active ingredients, particularly their toxicity, and provided a brief explanation.
Among the fve main active ingredients of Beishashen, stigmasterol, cnidilin, and alloisohyperperorin have not been reported in clinical research, animal experimental, or reviews related to their prevention and treatment of RILI and CILI.However, stigmasterol was mentioned in a network pharmacology article on adjuvant treatment of pneumonia and was discovered as a major component of the Feilike mixture [36].A popular phytosterol found in traditional Chinese medicine called beta-sitosterol has been shown to have anti-infammatory and antioxidant properties.At present, it has not been found to have a clear therapeutic efect in RILI and CILI, but some scholars have found that it can improve the proinfammatory reaction and acute lung injury induced by infuenza A virus in mice, as well as improve the lung histopathology injury and reduce the level of infammatory factors [37].Quercetin is the most abundant dietary favonoid found in various plants and foods, with strong anti-infammatory and antioxidant properties [38].Some researchers used quercetin as an intervention drug to preprotect RILI model-injured mice and found that in hematoxylin and eosin staining, the radiation control group showed more severe lung injury than the quercetin group [39].Immunohistochemistry and Western blotting results also revealed that, in contrast to the radiation control group, NF-B expression levels were reduced following quercetin intervention while NF-B inhibitor expression levels rose.In the radiation plus quercetin injection group, there were fewer infammatory cells that were JNK/ SAPK, p38, and p44/p42 positive (p < 0.05).Finally, they hypothesized that quercetin might inhibit NF-B and MAPK, which would protect mice's lungs from radiation.Meanwhile, Liu et al. also reckoned that the quercetin liposomes can prevent radiation-induced acute pneumonia and advanced fbrosis by reducing oxidative damage [40].Quercetin-3-rutinoside is a potential therapeutic agent that can alleviate radiation-induced lung injury in planned or unplanned radiation exposure situations [41].Although quercetin had certain protective efects and good therapeutic efects on lung injury caused by lipopolysaccharides, pneumolysin, bleomycin, paraquat, cigarette smoke, and even hyperoxia, there were few studies and reports on its use in CILI [42][43][44][45][46][47].Terefore, more investigation is required to determine whether quercetin protects against the lung damage brought on by chemotherapy medicines.
For decades, it has been well established that radiation produces reactive oxygen species (ROS) [8].However, chemotherapy, like radiotherapy, may induce endogenous production of ROS and nitric oxide by immune cells and some nonimmune cells (such as fbroblasts and endothelial cells) [48].After inhaling anticancer medications, the primary sources of ROS produced are cyclooxygenase-2 (PTGS2, also known as COX-2) and the mitochondrial cellular respiration system.ROS overexpression increases with the expression of PTGS2, leading to cell apoptosis,  [52].Besides, a clinical study had shown that genetic variations in the PI3K-AKT pathway are signifcantly associated with ≥3 levels of radiation pneumonia, which can serve as a predictive indicator for severe radiation pneumonia before radiotherapy [53].As shown in Figure 7, the MAPK signaling pathway is also regulated by the PI3K-AKT signaling pathway, so it may also be involved in the prevention and treatment mechanisms of RILI and CILI.Tis is also consistent with the research results of Arora and Xu et al. [54,55].Meanwhile, the TNF signaling pathway and the IL-7 signaling pathway, as classic pathways of infammatory response, also play important roles in RILI and CILI [55,56].With respect to molecular docking and molecular dynamics, the fve main active ingredients of Beishashen can bind to key targets, and their binding with energy is lower.Among them, the combination of alloisoimperatorin and RXRA is the best.In the further molecular dynamics simulation, we discovered that alloisoimperatorin and RXRA have a negative binding energy in the protein-ligand complex system, indicating that it is conducive to protein-ligand binding.Te binding free energy of alloisoimperatorin and RXRA is −23.437kJ•mol −1 , and there also exist hydrogen bonding interactions between alloisoimperatorin and RXRA, with an average number of hydrogen bonds of 1.39, showing that their binding is relatively tight.In addition, we also predicted the pharmacokinetic properties of these fve main active ingredients.According to the results of Table 4, quercetin, dnidilin, and alloisoimperatorin had an acute oral toxicity level of III, which is relatively low in toxicity.However, Cnidilin may have potential Ames mutagenesis, hepatotoxicity, and nephrotoxicity.Beta-sitosterol and stigmasterol had a level I acute oral toxicity, but no Ames mutagenesis, hepatotoxicity, and nephrotoxicity.Furthermore, these ingredients had no inhibitory efects on CYP2C19 and CYP3A4, but may have had potential inhibitory efects on other liver drug enzymes.At present, the traditional Chinese medicine prescription based on Beishashen has been applied in animal and clinical research, especially in the treatment and protection of radiation pneumonitis, which has achieved good curative efect, decreased infammatory factors, and the adverse reactions of patients.However, Beishashen has not been used alone for the time being.Terefore, Beishashen is mostly used in the form of prescriptions in the current clinical research and has achieved good results.In summary, quercetin and alloisoimperatorin may be reliable alternative substances for RILI or CILI prevention and treatment, but further experimental verifcation is also needed.More importantly, we must recognize that although bioinformatics analysis relies on public databases, the data information is imperfect and requires ongoing improvement.Furthermore, this study also overlooked the efects of Beishashen from diferent regions and the content and concentration of active ingredients on RILI and CILI because only a specifc number of medicines can efectively reach the target place.As a result, more confrmation and verifcation of the research's fndings are required.Self-made decoction Huangqin 6 g, Reed rhizome (Lugen) 6 g, Chuanbei 6 g, Lianqiao 6 g, Gancao 6 g, Huangjing 10 g, Xuanshen 10 g, Aster

Conclusions
To sum up, using network pharmacology, molecular docking, molecular dynamics, and big data information, our study investigated the probable mechanism of traditional marine Chinese medicine in preventing and treating RILI and CILI.As a result of the regulation of the PI3K-AKT signaling pathway, as well as the involvement of the PTGS2 and RXRA gene expression, which may serve as the main potential mechanisms of action, various components of Beishashen serve as various targets for the prevention and treatment of RILI and CILI.Tis study ofers not only a fresh look at the molecular mechanism of Beishashen for preventing and treating RILI and CILI, but it also serves as a foundation for contemporary pharmacology research in marine traditional Chinese medicine and even ofers guidance for future experimental studies of RILI and CILI.

Figure 1 :
Figure 1: Graphical synopsis representation of this study.
Figure3displays the fnal result.Inherent 129 nodes and 2107 edges with a PPI enrichment p value of 1.0e − 16 were included in the PPI network map, where nodes stood for proteins and edges for the innate connections between them.Te protein colors from orange to blue represent values from small to large.Te PPI network map provided a visual representation of the protein interaction relationships among the core targets.Te network can be further analyzed to identify key targets and pathways involved in the development and progression of RILI and CILI, which can help in the development of novel therapeutic strategies for these diseases.3.4.Development of the Active Ingredient-Disease-TargetNetwork.Te network diagram of Beishashen-ingredienttarget-RILI and CILI was constructed by Cytoscape 3.7.2software, as shown in Figure4.Te network revealed numerous interactions between active ingredients and key targets, suggesting that Beishashen can prevent and counteract RILI and CILI through multitarget and multipathway synergistic interactions.Five primary ingredients of Beishashen and fve important targets were examined using the Network Analyzer of the Cytoscape 3.7.2software.Te fve main active ingredients were quercetin, beta-sitosterol, stigmasterol, cnidilin, and alloisoimperatorin, while the fve key targets were prostglandinendoperoxide synthase 2 (PTGS2), sodium channel protein type 5 subunit alpha (SCN5A), phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit gamma (PIK3CG), prostglandinendoperoxide synthase 1 (PTGS1), and retinoic acid receptor RXR-alpha (RXRA).

Figure 3 :
Figure 3: PPI network of core key targets.

Figure 8 :
Figure 8: Te stability results from RXRA-alloisoimperatorin complex by MD simulations at 100 ns trajectory.(a) Protein RMSD and ligand RMSD with time frame in 100 ns; (b) simulation of protein-ligand complex process: changes in protein SASA in 100 ns; (c) simulation of protein-ligand complexation process: changes in Rg of protein in 100 ns; (d) simulation of protein-ligand complexation process: changes in protein RMSF in 100 ns; (e) changes in hydrogen bond numbers within 100 ns during protein-ligand recombination simulation process.

Table 1 :
Active ingredients and related targets from Beishashen.
Journal of Food Biochemistry 3.3.PPI Network Construction.After incorporating 129 core targets into the string platform and selecting "Homo sapiens" to generate the PPI network, and subsequently, associated protein interaction relationships were acquired.

Table 5 :
Te application of Beishashen and related formulas in RILI and CILI in the past 5 years.