Investigating the Active Substance and Mechanism of San-Jiu-Wei-Tai Granules via UPLC-QE-Orbitrap-MS and Network Pharmacology

San-Jiu-Wei-Tai granules (SJWTG) are a significant Chinese patent medicine for the treatment of chronic gastritis (CG), having outstanding advantages in long-term treatment; however, the chemical composition and potential mechanism have not been investigated until now. In this study, a rapid separation and identification method based on UPLC-QE-Orbitrap-MS was established, and 95 chemical components from SJWTGs were identified, including 6 chemical components of an unknown source that are not derived from the 8 herbs included in SJWTGs. The identified chemical components were subsequently analysed by network pharmacology, suggesting that the core targets for the treatment of CG with SJWTGs were EGFR, SRC, AKT1, HSP90AA1, MAPK1, and MAPK3 and thus indicating that SJWTGs could reduce the inflammatory response of gastric epithelial cells and prevent persistent chronic inflammation that induces cancerization by regulating the MAPK signalling pathway and the C-type lectin receptor signalling pathway as well as their upstream and downstream pathways in the treatment of CG. The key bioactive components in SJWTGs were identified as 2,6-bis(4-ethylphenyl)perhydro-1,3,5,7-tetraoxanaphth-4-ylethane-1,2-diol, a chemical component of an unknown source, murrangatin, meranzin hydrate, paeoniflorin, and albiflorin. The results of molecular docking showed the strong binding interaction between the key bioactive components and the core targets, demonstrating that the key bioactive components deserve to be further studied and considered as Q-markers. By acting on multiple targets, SJWTG is less susceptible to drug resistance during the long-term treatment of CG, indicating the advantage of Chinese patent medicines. Furthermore, the preventive effect of SJWTGs on gastric cancer also demonstrates the superiority of preventive treatment of disease with traditional Chinese medicine.


Introduction
Chronic gastritis (CG) is a disease involving chronic inflammatory or atrophic lesions of the gastric mucosa caused by various etiology [1], with clinical symptoms such as abdominal pain, fullness, burning sensation, acid reflux and belching, and loss of appetite as well as emotional changes like anxiety and depression. According to the new Sydney system for the classification of gastritis, CG is divided into chronic superficial gastritis (chronic non-atrophic gastritis) and chronic atrophic gastritis [2]. e incidence of CG is associated with a variety of factors, mainly including Helicobacter pylori (H. pylori) infection [3], duodenal-gastric reflux [4], drug factors [5], and autoimmunity [6]. H. pylori infection is the most common cause of CG, accounting for about 90%, and is also the main cause of gastric cancer [7]. However, the pathogenesis of CG has not been elucidated and remains to be further explored.
CG is the most common lifelong disease in human beings, possessing the characteristics of long duration and a strong chance of reoccurrence. It is estimated that more than half of the world's population suffers from CG, and millions of people worldwide die prematurely each year as a result of gastric cancer and gastric ulcer, which are the sequelae of CG [6]. In China, the prevalence of CG is the highest among digestive system diseases. In patients with long-lasting CG, the gastric epithelial cells are persistently infiltrated by inflammation, resulting in a variety of abnormal gene expressions and gene mutations [8]. Morphologically, prolonged inflammatory infiltration results in a thinning of the gastric mucosa, that is, gland atrophy, which indicates that chronic superficial gastritis has developed into chronic atrophic gastritis [2]. During the period of chronic atrophic gastritis, patients are so prone to gastric cancer that the World Health Organization has listed chronic atrophic gastritis as a pre-gastric cancer state [1].
Recently, research has shown that the pathogenesis of gastric cancer may be closely related to spasmolytic polypeptideexpressing metaplasia (SPEM). SPEM appears in the development of CG and represents the physiological healing response to injury [9], which also indicates that CG, as a pregastric cancer state, cannot be ignored. Nonetheless, most of the alterations can be reversed during the period of chronic superficial gastritis, suggesting that the timely and effective treatment of CG is of great significance.
Generally, Western medicine adopts triple (proton pump inhibitors, clarithromycin, and amoxicillin or metronidazole) or quadruple therapy (proton pump inhibitors, bismuth subcitrate or subsalicylate, tetracycline, and metronidazole) in treating CG. Nevertheless, there is still a risk of recurrence, drug resistance, and adverse reactions [10,11]. In contrast, Chinese patent medicines have outstanding advantages in the long-term treatment of CG due to their "multi-component and multi-target" characteristics. Among them, San-Jiu-Wei-Tai granules (SJWTGs) have been recommended for the treatment of CG in the clinical application guidelines on Chinese patent medicines for the treatment of CG (2020) [12]. SJWTG includes 8 herbs, Melicope pteleifolia (Champ. ex Benth.) T. G. Hartley, Murraya exotica L., Zanthoxylum nitidum (Roxb.) DC., Dolomiaea costus (Falc.) Kasana and A. K. Pandey, Scutellaria baicalensis Georgi, Poria cocos (Schw.) Wolf, Rehmannia glutinosa (Gaertn.) DC., and Paeonia lactiflora Pall., and is effective in clearing heat and dampness, promoting Qi and activating blood, softening the liver, and relieving pain. e ethanol extract of SJWTGs has been shown to treat anti-gastric ulcers, inhibit mucosal erosion and bleeding, and prevent large mucosal necrosis and shedding [13], but the mechanism remains unclear. To date, the chemical constituents of SJWTGs have not been systematically isolated and identified, and there has been no pharmacological evaluation of SJWTGs in treating CG. According to the Chinese Pharmacopoeia 2020 Edition, only baicalin is adopted as the Q-marker of SJWTGs [14], which makes quality control inadequate. erefore, it is urgent to investigate the chemical components of SJWTGs and explore the potential mechanism for treating CG.
Considering the "multi-component and multi-target" characteristics of traditional Chinese medicine (TCM), UPLC-QE-Orbitrap-MS was used to identify the chemical components in SJWTGs, due to its high resolution, high sensitivity, and high selectivity. Network pharmacology was then performed for insight into its potential mechanism in treating CG, including the key bioactive components, the core potential targets, and the vital signalling pathways. Finally, molecular docking was conducted to further verify the strong binding interactions between the key bioactive components and the core potential targets. e workflow of this study is illustrated in Figure 1. Our work lays a foundation for the further pharmacodynamics study of SJWTGs and guides clinical medication and quality control.

Preparation of SJWTG Extract.
Accurately weighed granules (1.00 g) were suspended in 10 mL 50% (v/v) methanol-water, ultrasonically extracted for 30 min, and then cooled to room temperature. After centrifuging at 10000 rpm for 10 min, 20 µL supernatant was diluted 50 times with 50% (v/v) methanol-water, and the diluted sample was then centrifuged at 12000 rpm for 10 min to obtain the final sample solution.

UPLC-QE-Orbitrap-MS Conditions.
We followed the methods of Feng et al. and the methods' description partly matches their wording [15]. Waters ACQUITY UPLC BEH C18 column (1.7 µm, 2.1 × 150 mm, Milford, MA, USA), column temperature 40°C, injection volume 3 µL, flow rate 0.3 mL/min, mobile phase comprising 0.1% formic acid aqueous solution (A) and acetonitrile (B). e eluting program was 5-5%B for 0-1 min, 5-95%B for 1-18 min, and 95-95%B for 18-20 min. e ion source was electron spray ionization (ESI); MS was operated in negative/positive mode; the scan mode was full scan/ddMS2, with positive and negative ion alternating scanning; and the scanning mode was 100-1300 Da, with a capillary temperature of 350°C. e spray voltage in positive mode was 3800 V, the spray voltage in negative mode was 3200 V, the sheath gas was 35 arb, and the auxiliary gas was 15 arb. ree collision energies of low, medium, and high were used for MS2. e positive ion mode was 20 V, 40 V, and 60 V, and the negative ion mode was 30 V, 50    with "chronic gastritis" as the keywords.

SJWTG-CG Common
Targets. SJWTG-CG common targets were obtained by taking the intersection of SJWTG targets and CG-related targets via a Venn diagram plotted by EHBIO (https://www.ehbio.com/).

PPI Network and Evaluation.
e SJWTG-CG common targets were imported into the STRING database (https://string-db.org/cgi/input.pl) to obtain proteinprotein interaction (PPI) information. e confidence score was set to 0.9 or higher, and the edge free proteins were excluded in order to ensure the reliability of the research data. Cytoscape 3.7.2 software was used to visualize the structure of the protein network and analyse its topological characteristics. e core targets for the treatment of CG with SJWTGs were obtained by taking the median of degree centrality (DC), betweenness centrality (BC), and closeness centrality (CC), as screening conditions, twice.

GO Analysis and KEGG Enrichment Analyses.
e core targets for the treatment of CG with SJWTGs were imported into Metascape (https://metascape.org/) for gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses, and the enrichment diagram was plotted by bioinformatics.

Core Component-Target-Pathway Network.
e effective components, core targets, and action pathways of SJWTGs were imported into Cytoscape 3.7.2 software to establish the core component-target-pathway network, with the purpose of mining the key bioactive components and vital pathways. KEGG Mapper (https://www.kegg.jp/kegg/ mapper/) was then used to annotate the roles of important targets of the vital pathways.

Molecular Docking.
e two-dimensional (2D) structures of the compounds used as ligands were obtained from the PubChem database (https://pubchem.ncbi.nlm.nih.gov/ ), processed to minimize energy, and transformed into the MOL format as ligands for docking through Chem3D. e X-ray crystal structures of the proteins were obtained from the Protein Data Bank (PDB, https://www.rcsb.org/). Afterwards, the structures of these proteins were optimized by Maestro 11.8 software, including the assignment of bonds and bond orders, the addition of hydrogens, filling in missing loops or side chains, capping uncapped termini, adjusting bonds and formal charges for metals, correcting mislabelled elements, and deleting unnecessary parts in the structure. Subsequently, each grid box was centred on the original ligand in each protein. e docking results were visualized and displayed as three-dimensional (3D) diagrams and 2D diagrams using Maestro 11.8.

Chemical Component Analysis.
e retention time and mass spectrometry information of the compounds from SJWTGs were obtained by UPLC-QE-Orbitrap-MS. Due to the complexity of chemical components, the SJWTG samples were fully scanned under positive and negative ionization modes. e base peak intensity chromatogram (BPI) is shown in Figure 2. Based on accurate mass spectrometry information, such as fragment ions, and reference literature, 95 chemical components were identified in SJWTGs (Table 1) and were categorized into 9 classes of natural compounds ( Figure 3). e chemical structures of 95 identified compounds are shown in Supplementary Materials ( Figure S1).     ucdavis.edu/), compound 60 was identified as marmesin ( Figure S2).   us, compound 75 was identified as costunolide by consulting MassBank Europe (https://massbank.eu/MassBank/) ( Figure S5).  e rectangle nodes stand for chemical components of an unknown source. e brown triangular nodes, the blue triangular nodes, the green triangular nodes, the purple triangular nodes, the yellow triangular nodes, the red triangular node, and the orange triangular nodes stand for unique components of Melicope pteleifolia, Murraya exotica, Zanthoxylum nitidum, Dolomiaea costus, Scutellaria baicalensis, Poria cocos, Rehmannia glutinosa and Paeonia lactiflora respectively. e colour scales and the size of the nodes represent the degree level of components and targets.  +H] + by the neutral loss of NH 3 . As the MS2 spectra of this compound exhibited a high relative abundance of ions at m/z 110.0351 and 93.0578, the first fragmentation pathway was shown to be dominant, which is the characteristic of the cleavage of purine. Moreover, the fragment ions at m/z 110.0351, 109.517, and 135.0301 suggested that the substituent groups of CO and NH 2 were located in the pyrimidine ring. erefore, compound 8 was identified as guanine by consulting mzCloud ( Figure S7).       (Figure 4). e colour scales and the size of the nodes represent the degree level of the components and targets. Among these nodes, 780 targets were from the unique components of the 8 herbs, 372 targets were from the common components, and 202 targets were from chemical components of an unknown source. Considering the aesthetic of the networks in this part, abbreviated names were given to the 95 identified compounds, as shown in the Supplementary Materials (Table S1).

Acquisition of SJWTG-CG Common Targets.
Using the keyword "chronic gastritis" to search for disease targets in OMIM and GeneCards, and screening and merging to remove duplicates, a total of 1265 CG-related targets were obtained. A Venn diagram was plotted by EHBIO, and 225 SJWTG-CG common targets were obtained ( Figure 5).

Construction and Evaluation of PPI Network.
e STRING database was employed to predict the PPIs of the 225 common targets. In this study, medium confidence >0.9 was set, edge free proteins were excluded, and the reliability of the research data was ensured. e topological characteristics of the protein network structure were analysed by Cytoscape 3.7.2 software, and 17 core targets for the treatment of CG with SJWTGs were screened, including SRC, STAT3, MAPK3, HSP90AA1, MAPK1, and AKT1 ( Figure 6 and Table S2). e colour scales and the size of the circles represent the degree level of these target proteins.

GO and KEGG Enrichment Analyses.
To further investigate the core targets of SJWTGs in treating CG, GO analysis was carried out using Metascape, and biological process (BP), molecular function (MF), and cellular components (CCs) were screened out. e drawing was made using bioinformatics ( Figure 7). e top 10 in the BP analysis are relevant to the cellular response to stimulus and the positive regulation of cell motility and migration, the top 10 in the CC analysis are related to the biomembrane system, and the top 10 in the MF analysis reveal that the core targets could affect the activity and binding of various phosphatases and kinases.
Metascape was employed to explore the KEGG pathway information that common targets may participate in. As a result, a total of 139 signal pathways were identified, and the top 10 pathways including distinctive genes were selected (Figure 8). e mechanism of SJWTGs in treating CG mainly involves the "C-type lectin receptor signalling pathway," "EGFR tyrosine kinase inhibitor resistance," and "MAPK signalling pathway." e results of KEGG pathway analyses correspond to the results of the GO pathway analyses, indicating that the core targets of SJWTGs are related to the cellular response to stimulus by affecting the activity 18 Evidence-Based Complementary and Alternative Medicine and binding of various phosphatases and kinases, which might be consistent with the treatment of CG with SJWTGs.

Construction of Core Component-Target-Pathway
Network. With the purpose of gaining a holistic understanding of the underlying mechanism of SJWTGs, a component-target-pathway network was constructed by Cytoscape 3.7.2 software, including 94 nodes and 255 edges. Overall, 56 unique components of the 8 herbs, 13 common components, and 4 components of an unknown source participate in the regulation of 16 core targets and 10 pathways. e colour scales and the size of the nodes represent the degree level of the components, targets, and pathways. As shown in Figure 9, 2,6-bis(4-ethylphenyl) perhydro-1,3,5,7-tetraoxanaphth-4-ylethane-1,2-diol (NC3), murrangatin (JLX4), meranzin hydrate (JLX5), paeoniflorin (BS9), and albiflorin (BS7) are of great importance in treating CG. e top 10 essential targets in the component-target-pathway network are EGFR, SRC, MAPK14, AKT1, MAPK8, HSP90AA1, TNF, MAPK1, MAPK3, and VEGFA. Meanwhile, SRC, STAT3, MAPK3, HSP90AA1, MAPK1, AKT1, HRAS, EGFR, JUN, and IL6 are also vital targets in the PPI network. e duplicated targets between these two networks are EGFR, SRC, AKT1, HSP90AA1, MAPK1, and MAPK3, which are the core targets of SJWTGs in treating CG. Subsequently, we found that all of these targets play important roles in the MAPK signalling pathway and the C-type lectin receptor signalling pathway, suggesting that these two pathways are vital signalling pathways during the process by which SJWTG treats CG. KEGG Mapper was used to annotate the roles of the important targets of the two vital signalling pathways ( Figure 10).

Molecular Docking.
Based on the results of network pharmacology predictions, we believed that 2,6-bis(4-ethylphenyl)perhydro-1,3,5,7-tetraoxanaphth-4-ylethane-1,2diol, murrangatin, meranzin hydrate, paeoniflorin, and albiflorin were the key bioactive components, while EGFR, SRC, AKT1, HSP90AA1, MAPK1, and MAPK3 were the core targets. ese proteins not only play an important role in the KEGG signalling pathways but also serve as the key nodes of the PPI network and core component-targetpathway network. erefore, the key bioactive components were used as ligands, and the core targets were used as proteins for the molecular docking, in order to further verify the network pharmacology results. As shown in Table 2, the binding energies were computed to evaluate the binding affinities of the 5 compounds with the 6 proteins, respectively. It is commonly recognized that a more negative binding energy value indicates a stronger binding affinity or a greater binding constant for the formation of the ligand-protein complex. e binding energies of the 5 compounds with the 6 proteins ranged from −4.698 to −7.876 kcal/mol, suggesting a degree of confidence in the network pharmacology results, namely that these 5 compounds may have potential therapeutic effects on CG by binding to the 6 proteins. Among them, EGFR and SRC have the lowest binding energy with murrangatin; AKT1 and MAPK3 have the lowest binding energy with meranzin hydrate; and HSP90AA1 and MAPK1 have the lowest binding energy with paeoniflorin. e 3D and 2D action mode graph of the representative compounds and related proteins are shown in Figure 11 and the results, displayed through a heat map, are shown in Figure 12

Evidence-Based Complementary and Alternative Medicine 21
paeoniflorin is a nonsteroidal anti-inflammatory agent, which can modulate the activation of immune cells and decrease inflammatory medium production via regulating the MAPK/NF-κB pathway, JAK2/STAT3 pathway, and PI3K/Akt/mTOR pathway [23]. Meanwhile, albiflorin could regulate the MAPK/NF-κB signalling pathway by inhibiting the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), TNF-α, and IL-6, thereby acting as an anti-inflammatory [24,25]. We found that the core targets of SJWTGs in treating CG were EGFR, SRC, AKT1, HSP90AA1, MAPK1, and MAPK3. Gastric epithelial EGFR inhibition represents a potential strategy to prevent the development of gastric cancer in H. pylori-infected individuals [26]. Human gastric tissues exhibit elevated levels of phosphorylated EGFR during the progression from CG to gastric cancer, which could activate the NF-κB and MAPK1/3 pathways to induce cytokine production and macrophage activation, while EGFRdeficient macrophages display impaired 1 and 17 adaptive immune responses to H. pylori [27]. SRC plays an important role in the migration of gastric cancer cells by mediating a potential CXCR4-EGFR crosstalk and sequentially activating the EGFR-Akt/ERK axis [28]. Additionally, SRC has been proven to be the kinase mediating the phosphorylation of CagA, a protein delivered into the bacterium-attached gastric epithelial cell by H. pylori, in vitro and in vivo [29]. Studies have found that AKT1 gene mutation could drive pathogenicity in gastric cancer [30], and the proliferation and apoptosis of gastric cancer cells could be regulated by affecting the expression level of AKT1 [31,32]. HSP90AA1, as the target of miR-9-5p, could be targeted to suppress gastric cancer cell proliferation and metastasis [33], and the MAPK1/3 pathway has played crucial roles in H. pylori infection, CG, and gastric cancer [26,27]. Several studies have shown that the tumour growth of gastric cancer could be restrained by inhibiting the expression level of MAPK1 and MAPK3 [34][35][36][37]. Moreover, STAT3, HRAS, MAPK14, and MAPK8 are also important in the process of SJWTGs treating CG: pS-STAT3 is essential during the entire pathologic progression from CG to gastric cancer in H. pylori-infected mice [38]; HRAS, markedly upregulated in gastric cancer, could enhance the aggressiveness of gastric cancer by activating the VEGFA/PI3K/ AKT pathway and Raf-1 signalling [39]; MAPK14 could be a potential biomarker for advanced gastric cancer, as well as a pharmacological target, which could improve the survival rate of patients [40]; and the upregulation of MAPK8 expressed in gastric cancer tissues is relevant to a switch towards a premalignant state in H. pylori-infected tissues [41].
Furthermore, we suggest that the MAPK signalling pathway and C-type lectin receptor signalling pathway are vital during the treatment of CG by SJWTGs. e MAPK signalling pathway could be activated by H. pylori infection to induce cytokine production and macrophage activation, resulting in the formation of CG [27], which also seems to be a gold target for anticancer therapies, especially for gastric cancer [42]. During the development of H. pylori-induced CG into early gastric cancer, Lewis antigens of H. pylori LPS could interact with Mincle (Macrophage inducible C-type lectin) and maintain a balance between pro-and antiinflammatory cytokine production, suggesting that the Ctype lectin receptor signalling pathway is closely related to the mechanism employed by H. pylori to escape the host innate immune receptors [43]. In addition, the EGFR tyrosine kinase inhibitor resistance, TNF signalling pathway, HIF-1 signalling pathway, 17 cell differentiation, T-cell receptor signalling pathway, Toll-like receptor signalling pathway, IL-17 signalling pathway, and ErbB signalling pathway have also played crucial roles in the treatment of CG by SJWTGs. e expression of EGFR tyrosine kinase is highly deregulated in gastric cancer tissues, as a result of H. pylori [44], indicating that EGFR tyrosine kinase inhibitor resistance could regulate the imbalance of gastric cancer. TNF-α can increase the release of pro-inflammatory cytokines, augmenting apoptosis induced by H. pylori [45], while monoclonal antibodies targeting TNF-α have been investigated for their potential to prevent inflammation-based gastric cancer [46]. As major transcriptional regulators of immunity and inflammation, HIFs are closely related to CG. Studies have found that HIF-1 is protective in H. pylorimediated CG, indicating that the HIF-1 signalling pathway has a potential therapeutical effect [47]. H. pylori induces 17 cell differentiation via infected macrophages, and the number of 17 cells is positively correlated with the degree of CG [48,49]. e T-cell receptor signalling pathway could be activated by H. pylori protein HP1454, leading to an inflammatory response [50]. Moreover, H. pylori infection could initiate chemokine-mediated T lymphocyte trafficking into the inflamed epithelium and induce mucosal injury [51]. Toll-like receptor 4 plays an important role in the Tolllike receptor signalling pathway, which could mediate an inflammatory response in the gastric epithelia induced by H. pylori and sequentially contribute to the initiation and progression of gastric cancer cells [52]. As a hallmark cytokine of 17 cells, IL-17 plays a critical role in the host's defence against bacterial infection [53]. H. pylori infection perturbs the balance between T-regulatory and 17 cells, which causes a spurt of IL-17 and gives rise to CG [54]. Gastric cancer exhibits alterations in the ErbB receptor family and ErbB-related signalling pathways [55]; in particular, the expression level of ErbB2 is increased in gastric cancer, and ErbB2 targeted therapies for gastric cancer have proved highly beneficial [56]. e results of molecular docking showed the strong binding interaction between 5 key bioactive components and 6 core targets, which further verifies the network pharmacology results, to a certain degree. Due to the structural similarity of the 5 key bioactive components, that is, the existence of lactones, hydroxyl groups, and phenyl groups in the structure, they all have good conjugation with the core targets via polar interactions and hydrophobic interactions. Consequently, we suggest that the 5 key bioactive components could be considered as Q-markers. Nevertheless, molecular docking is still based on computer simulation, and the results need to be validated in vivo.
erefore, we believe that SJWTGs could reduce the inflammatory response of gastric epithelial cells and prevent long-term chronic inflammation to induce cancerization by regulating the MAPK signalling pathway and the C-type lectin receptor signalling pathway as well as their upstream and downstream pathways. is is different to the triple and quadruple therapies that use antibiotics to eradicate H. pylori, indicating that SJWTG is also curative in the treatment of CG without H. pylori infection. Moreover, we conclude that the pathogenesis of CG may be related to the disorder of the MAPK signalling pathway and the C-type lectin receptor signalling pathway, and that H. pylori infection, duodenalgastric reflux, drugs, and autoimmunity are only the key factors to induce the disorder of these pathways. Furthermore, the preventive effect of SJWTGs on gastric cancer also shows the superiority of preventive treatment of diseases with TCM. Due to the variety of targets, including EGFR, SRC, AKT1, HSP90AA1, MAPK1, and MAPK3, SJWTG is less susceptible to drug resistance during the long-term treatment of CG, suggesting the advantage of Chinese patent medicines. Additionally, 2,6-bis(4-ethylphenyl)perhydro-1,3,5,7-tetraoxanaphth-4-ylethane-1,2-diol, murrangatin, meranzin hydrate, paeoniflorin, and albiflorin have been shown to be the key bioactive components of SJWTGs in treating CG: the results of molecular docking demonstrate their potential therapeutic effects on CG and these components deserved to be further studied and considered as Q-markers, especially the chemical component of an unknown source that is not derived from the 8 herbs, 2,6-bis(4-ethylphenyl)perhydro-1,3,5,7-tetraoxanaphth-4-ylethane-1,2-diol.

Conclusions
In summary, a comprehensive approach based on UPLC-QE-Orbitrap-MS and network pharmacology has been used to explore the active compounds and potential mechanism of SJWTGs in treating CG. Overall, 95 chemical components were identified by UPLC-QE-Orbitrap-MS, including 6 chemical components of an unknown source that are not derived from the 8 herbs present in SJWTGs. Subsequently, the identified chemical components were further researched by network pharmacology, where it was found that SJWTGs could reduce the inflammatory response of gastric epithelial cells and prevent persistent chronic inflammation to induce cancerization by regulating the MAPK signalling pathway and the C-type lectin receptor signalling pathway, as well as their upstream and downstream pathways, in the treatment of CG, which could also be related to the pathogenesis of CG.
e key bioactive components are 2,6-bis(4-ethylphenyl) perhydro-1,3,5,7-tetraoxanaphth-4-ylethane-1,2-diol, a chemical component of an unknown source, murrangatin, meranzin hydrate, paeoniflorin, and albiflorin. e results of molecular docking showed the strong binding interaction between the key bioactive components and the core targets, demonstrating that the key bioactive components deserved to be further studied and considered as Q-markers. Acting on multiple core targets, including EGFR, SRC, AKT1, HSP90AA1, MAPK1, and MAPK3, SJWTG is less susceptible to drug resistance during the long-term treatment of CG, suggesting the advantage of Chinese patent medicines. Furthermore, the preventive effect of SJWTGs on gastric cancer also demonstrates the superiority of the preventive treatment of disease with TCM. Figure S1: e chemical structures of 95 identified compounds. Figure S2: e MS/MS spectrum of marmesin in positive ion mode and probable fragmentation pathway. Figure S3: e MS/MS spectrum of murralongin in positive ion mode and probable fragmentation pathway. Figure S4: e MS/MS spectrum of nitidine in positive ion mode and probable fragmentation pathway. Figure S5: e MS/MS spectrum of costunolide in positive ion mode and probable fragmentation pathway. Figure S6: e MS/MS spectrum of scutellarin in positive ion mode and probable fragmentation pathway. Figure S7: e MS/MS spectrum of guanine in positive ion mode and probable fragmentation pathway. Figure S8: e MS/MS spectrum of acteoside in negative ion mode and probable fragmentation pathway. Figure S9: e MS/MS spectrum of albiflorin in positive ion mode and probable fragmentation pathway. Figure S10: e MS/MS spectrum of 2,6-bis(4-ethylphenyl)perhydro-1,3,5,7-tetraoxanaphth-4-ylethane-1,2-diol in positive ion mode and probable fragmentation pathway. Table S1: Abbreviated names of 95 identified compounds.