Identifying the active compounds and mechanism of action of Banxia Xiexin decoction for treating ethanol-induced chronic gastritis using network pharmacology combined with UPLC–LTQ–Orbitrap MS

https://doi.org/10.1016/j.compbiolchem.2021.107535Get rights and content

Highlights

  • UPLC-MS/MS showed the active compounds of BXD for treating gastritis.

  • Network pharmacology uncovered the mechanism of BXD in treating gastritis.

  • SPR assessed the affinity between the active ingredients and potential targets.

  • ICAM-1, PPAR-γ, and MAPK14 might become novel drug targets for gastritis.

Abstract

Background

Banxia Xiexin decoction (BXD), a traditionally prescribed Chinese medicine, has been used to treat chronic gastritis for many years. However, the underlying mechanism and targets for its effects remain unknown. In the present study, we predicted the targets and active compounds of BXD in the treatment of chronic gastritis through network pharmacology and ultra-performance liquid chromatography coupled with linear trap quadrupole–Orbitrap mass spectrometry (UPLC–LTQ–Orbitrap MS).

Method

A chronic gastritis model was established in rats by oral administration of 56 % ethanol. BXD was orally administered for 7 days. Stomach tissues were collected for histopathological analysis, and tumour necrosis factor (TNF)-α, interleukin (IL)-2, IL-8, and lactate dehydrogenase (LDH) levels were measured by enzyme-linked immunosorbent assay. UPLC–LTQ–Orbitrap MS was established to analyse compounds in rat plasma following oral BXD administration. The absorbed ingredients were selected as candidate active compounds. The chronic gastritis-related targets were screened using multiple databases. The potential targets for the treatment of chronic gastritis were used to construct a protein–protein interaction (PPI) network and were also analysed using the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. Finally, molecular docking was used to uncover the interaction between multi-components and putative targets, and the results were verified by surface plasmon resonance (SPR).

Results

Intragastric administration of BXD ameliorated stomach injury resulting from chronic gastritis in rats and decreased the levels of TNF-α, IL-2, IL-8, and LDH. A comprehensive systematic strategy was used to successfully identify 38 candidate targets and 14 active compounds in BXD. Based on the network of compounds–targets and PPI, three hub genes that were associated with BXD therapy for chronic gastritis were selected and included intercellular adhesion molecule-1, peroxisome proliferator-activated receptor gamma and mitogen-activated protein kinase 14. The results of molecular docking and SPR demonstrated that the active compounds in BXD demonstrate affinity for these targets. Additionally, an enrichment analysis revealed that treatment of chronic gastritis with BXD primarily involves cytokine activation, the inflammatory response and nuclear factor-kappa B, hypoxia-inducible factor-1, phosphatidylinositol-3-kinase–protein–serine–threonine kinase and Janus kinase–signal transducer and activator of transcription signalling pathways, which may mediate the effects of BXD in the treatment of chronic gastritis.

Conclusion

BXD exhibits a therapeutic effect in ethanol-induced gastritis through multi-compound, multi-target and multi-pathway mechanisms. A strategy of network pharmacology combined with SPR may provide a feasible approach to explore the targets of herbal medicine and uncover novel bioactive components.

Introduction

Gastritis is an inflammatory disease that includes injury to the gastric mucosa. It is typically associated with symptoms that include stomach pain, bloating and anorexia. Chronic gastritis has become a major threat to human health and is predominantly characterised by mononuclear inflammatory cells (Estevam et al., 2017). Excessive or chronic drinking of strong alcohol can induce gastric mucosa congestion, oedema and erosion, as well as acute or chronic gastritis (Bode and Bode, 1997; Shin et al., 2013). Alcohol may be converted to acetaldehyde and induce local toxicity in the gastrointestinal tract (Seitz et al., 1994). Meanwhile, inflammatory cytokines including tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-2, and IL-8 are induced by ethanol (Ghare et al., 2011; Tanyeli et al., 2017). IL levels significantly increase the production of intercellular adhesion molecules (ICAMs) after alcohol ingestion, and there is a close relationship with serum ICAM-1 levels and alcohol intake (Sacanella et al., 1999). Lactate dehydrogenase (LDH) is significantly activated by aspirin, induces acute gastritis and contributes to the development of gastric lesions (Du et al., 2014). The inhibition of caspase-3 cleavage and the regulation of phosphatidylinositol-3-kinase (PI3K)/protein–serine–threonine kinase (Akt) transduction, nuclear factor-kappa B (NF-κB) activation and mitogen-activated protein kinase (MAPK) signalling play key roles in the process of gastric mucosal injury (Arab et al., 2019).

Banxia Xiexin decoction (BXD) has been used clinically for thousands of years, and it shows the characteristics of an effective multi-component, multi-target therapeutic with few side effects (Chen et al., 2015a,b). This formula is widely used to treat gastric motility disorders, gastritis, colitis and peptic ulcer by acting on the spleen and stomach (Lu et al., 2018). The prescription consists of several medicinal herbs or their parts (Pinellia Rhizoma, Glycyrrhizae Radix et Rhizoma, Scutellaria Radix, Coptidis Rhizoma, Ginseng Radix et Rhizoma, Zingiberis Rhizoma and Jujube Fructus). Previous studies showed that BXD contains multiple active components including berberine, baicalin, baicalein, wogonin, wogonoside, coptisine, palmatine, and ginsenosides (Wang et al., 2014; Yan et al., 2013). Berberine, which attenuates the B cell-activated factor-triggered Th17 response, exerts anti-inflammatory effects on Helicobacter pylori-induced chronic gastritis (Wu and Li, 2018). Baicalin and baicalein promote health by preventing H. pylori infection and interfering with H. pylori growth and virulence (Chen et al., 2018a,b). However, there are many complex ingredients in BXD, and its mechanism of action and targets for treating various diseases remains obscure.

Network pharmacology is a method used to study the regulation of diseases by multi-component drugs (Zhang et al., 2019; Zhu et al., 2018). It integrates system bioinformatics and omics methods to identify target proteins and signalling pathways associated with drug efficacy from an integral perspective and incudes characteristics of completeness, synergy and dynamics (Lim and Xie, 2019; Wu et al., 2018). Furthermore, surface plasmon resonance (SPR) technology may be applied to traditional Chinese Medicine (TCM), and high-throughput screening of bioactive components can be used to identify its multi-component chemicals. The most important advantage of SPR is the ability to monitor the dynamic process of biological macromolecules with compounds in real time (Aristotelous et al., 2015; Lago et al., 2018).

Consequently, the aim of the present study was to use an innovative strategy to identify the molecular mechanisms and potential targets of BXD using a systematic pharmacological approach, SPR, molecular mechanics simulation and experimental verification to gain insight into the therapeutic properties of BXD in treating ethanol-induced chronic gastritis.

Section snippets

Reagents and materials

Pinellia Rhizoma, Scutellaria Radix, Ginseng Radix et Rhizoma, Zingiberis Rhizoma, Coptidis Rhizoma, Glycyrrhizae Radix et Rhizoma and Jujube Fructus were selected based on the standards specified in the Chinese Pharmacopoeia and were purchased from the Shanghai Kang Qiao Chinese Medicine Tablet Co., Ltd. Wogonoside, wogonin, berberine, baicalin, baicalein, and glycyrrhetinic acid were purchased from Yuanye Biological (Shanghai, China). Enzyme-linked immunosorbent assay (ELISA) kits for IL-2,

Effects of BXD on ethanol-induced chronic gastritis in rats

The gastric mucosa of the normal group was not damaged. In contrast, erosion, ulcers and bleeding of the gastric mucosa were observed in the model group. The gastric mucosal damage of the BXD group was lower than that of the model group (Fig. 2A). H&E staining of the stomach tissues revealed that there was inflammation in the infiltrate of immune cells in the model group. Oedema and congestion were also apparent in the submucosal and intrinsic layers as well as incomplete submucosa. However,

Discussion

Chronic gastritis is the result of damage and inflammation in the gastric mucosa. Ethanol may disrupt the gastric mucosal barrier defence system and reduce the ability of the gastric mucosa to defend against the effects of gastric acid, leading to mucosal oedema, erosion, haemorrhage and necrosis (Ning et al., 2012). Treatment for alcoholic gastritis is limited, and some adverse reactions remain a major challenge (Bandyopadhyay et al., 2002). BXD has been used for the treatment of chronic

Conclusions

Based on the analysis of the serum components of BXD by molecular docking and compound–target network analysis, some active ingredients and target proteins were selected for SPR validation. KEGG pathway and PPI network analysis indicated that the NF-κB, HIF-1, PI3K-Akt, and JAK-STAT signalling pathways participate in the occurrence and treatment of ethanol-induced chronic gastritis. Furthermore, molecular docking and SPR methods were used to validate the interactions between key compounds and

Author statement

WJ wrote the original draft. KL performed the experiments. TW and YX analyzed the data. RA revised the manuscript. XW conceived of the study and designed the study. All authors read and approved the final version of the manuscript.

Funding

This work was supported by National Natural Science Foundation of China (grant No.81774183), and sponsored by Scientific Research Foundation of Shanghai University of Medicine and Health Sciences (grant number E3-0200-21-201011-34).

Declaration of Competing Interest

The authors report no declarations of interest.

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