Exploration of the mechanism of Danggui Buxue decoction (当归补血汤) for the treatment of gastric ulcer based on network pharmacology, molecular docking, and in vivo experiment

doi:10.19852/j.cnki.jtcm.20250318.002

Abstract

Abstract:

OBJECTIVE: To explore the mechanism of Danggui Buxue decoction (当归补血汤, DBD) for the treatment of gastric ulcer (GU), based on network pharmacology and in vivo experiments.

METHODS: A network pharmacology strategy was used to predict the main components, candidate targets, and potential signaling pathways. Then, molecular docking was performed to further investigate the interactions and binding affinities between the main components and primary targets. Finally, a mouse model of ethanol-induced gastric ulcers was established to confirm the efficacy and potential therapeutic benefits of DBD, and candidate targets were finally identified.

RESULTS: A total of 22 active components and 220 target genes were found to be associated with DBD. In addition, 343 GU-related target genes and 57 target genes specific to DBD treatment of GU were identified. The Gene Ontology functional enrichment analysis revealed 510 entries for biological processes, 36 entries for cell composition, and 69 entries for molecular functions. In the pathway enrichment analysis, 143 signaling pathways were identified. Additionally, the molecular docking results revealed that the main active components of DBD exhibited a strong binding capacity with key proteins, including tumor necrosis factor, AKT serine/threonine kinase 1, interleukin-6, vascular endothelial growth factor, and interleukin-1 Beta. Among these, quercetin, kaempferol, formononetin, isorhamnetin, and beta-sitosterol displayed the strongest binding affinities for these key proteins. in vivo experiments showed that DBD pretreatment effectively protected gastric mucosa, and the benefits might be attributed to the downregulation of above key proteins.

CONCLUSIONS: Based on network pharmacology analysis and in vivo experiments, we conclude that DBD leads to the protection and healing of the gastric mucosa by targeting genes and pathways, thus effectively countering the development and progression of GU.

Key words: stomach ulcer, network pharmacology, molecular docking simulation, mechanism, Danggui Buxue decoction

SONG Mingming, MEN Bo, CHEN Mei, LIU Rui, MO Hongping, ZHANG Da, PAN Tao, WEN Xudong. Exploration of the mechanism of Danggui Buxue decoction (当归补血汤) for the treatment of gastric ulcer based on network pharmacology, molecular docking, and in vivo experiment[J]. Journal of Traditional Chinese Medicine, 2025, 45(4): 806-816.

Figures/Tables 6

Table 1 Active ingredients of Danggui Buxue decoction

Herbs	Mol ID	Molecule name	OB (%)	DL
Danggui (Radix Angelicae Sinensis)	MOL000358	Beta-sitosterol	36.91	0.75
	MOL000449	Stigmasterol	43.83	0.76
Huangqi (Radix Astragali Mongolici)	MOL000211	Mairin	55.38	0.78
	MOL000239	Jaranol	50.83	0.29
	MOL000296	Hederagenin	36.91	0.75
	MOL000033	(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-ol	36.23	0.78
	MOL000354	Isorhamnetin	49.6	0.31
	MOL000371	3,9-di-O-methylnissolin	53.74	0.48
	MOL000374	5'-hydroxyiso-muronulatol-2',5'-di-O-glucoside	41.72	0.69
	MOL000378	7-O-methylisomucronulatol	74.69	0.3
	MOL000379	9,10-dimethoxypterocarpan-3-O-β-D-glucoside	36.74	0.92
	MOL000380	(6aR,11aR)-9,10-dimethoxy-6a,11a-dihydro-6H-benzofurano[3,2-c]chromen-3-ol	64.26	0.42
	MOL000387	Bifendate	31.1	0.67
	MOL000392	Formononetin	69.67	0.21
	MOL000398	Isoflavanone	109.99	0.3
	MOL000417	Calycosin	47.75	0.24
	MOL000422	Kaempferol	41.88	0.24
	MOL000433	FA	68.96	0.71
	MOL000438	(3R)-3-(2-hydroxy-3,4-dimethoxyphenyl) chroman-7-ol	67.67	0.26
	MOL000439	Isomucronulatol-7,2'-di-O-glucosiole	49.28	0.62
	MOL000442	1,7-Dihydroxy-3,9-dimethoxy pterocarpene	39.05	0.48
	MOL000098	Quercetin	46.43	0.28

Figure 1 Network pharmacology analysis of DBD on GU A: PPI network for 57 potential targets; B: PPI network for the 14 key targets. DBD: Danggui Buxue decoction; GU: gastric ulcer; PPI: protein-protein interaction.

Table 2 Details of the key targets

Key targets	Betweenness centrality	Closeness centrality	Degree
TNF	0.07224798	0.65882353	33
IL-6	0.06024239	0.65116279	33
VEGF	0.09232003	0.66666667	31
AKT1	0.1051178	0.64367816	28
TP53	0.08365778	0.62921348	26
MMP9	0.07389262	0.62222222	26
IL-1β	0.01989997	0.58947368	25
CASP3	0.0375621	0.57731959	21
RELA	0.03861559	0.57142857	21
EGF	0.03017289	0.56565657	20
MYC	0.05219103	0.56	17
PTGS2	0.0320954	0.54368932	17
MAPK1	0.0283965	0.54368932	16
MMP2	0.0200976	0.53846154	14

Figure 2 Molecular docking: The hydrogen bond lengths, amino acid residues, and the active pockets between active components and proteins A: TNF-quercetin; B: TNF-kaempferol; C: IL-6-quercetin; D: VEGF-quercetin. TNF: tumor necrosis factor; IL-6: interleukin-6, VEGF: vascular endothelial growth factor.

Figure 3 Effect of DBD on gastric lesions induced by ethanol A: macroscopic examination of the gastric mucosa in mice of different groups; B: the average ulcer index of stomach samples in each group; C: results of HE staining of gastric mucosa (scale bar = 100 μm). A1, C1: Control group; A2, C2: Model group; A3, C3: Ranitidine group; A4, C4: DBD-H group; A5, C5: DBD-M group; A6, C6: DBD-L group. Control (distilled water, 1 week); Model (distilled water, 1 week); Ranitidine (ranitidine, 30 mg/kg body weight, 1 week); DBD-H (DBD, 2.34 g/kg, dry weight, 1 week); DBD-M (DBD, 1.17 g/kg, dry weight, 1 week); DBD-L (DBD, 0.59 g/kg, dry weight, 1 week). Statistical analyses were measured using one-way analysis of variance for multimal comparisons. Data were presented as mean ± standard deviation (n = 6). aP < 0.001 vs control group; bP < 0.001, cP < 0.05 vs model group. DBD: Danggui Buxue decoction.

Figure 4 Comparison of protein expression and mRNA expression levels A: representative results of protein phosphorylation levels in each group; B: quantitative results of protein phosphorylation levels in each group; C: relative mRNA expression of TNF-α; D: relative mRNA expression of IL-6; E: relative mRNA expression of VEGF; F: relative mRNA expression of IL-1β; G: relative mRNA expression of AKT1. Control (distilled water, 1 week); Model (distilled water, 1 week); Ranitidine (ranitidine, 30 mg/kg body weight, 1 week); DBD-H (DBD, 2.34 g/kg, dry weight, 1 week); DBD-M (DBD, 1.17 g/kg, dry weight, 1 week); DBD-L (DBD, 0.59 g/kg, dry weight, 1 week). DBD: Danggui Buxue decoction; TNF: tumor necrosis factor; AKT1: AKT serine/threonine kinase 1; IL-6: interleukin-6, VEGF: vascular endothelial growth factor; IL-1β: interleukin-1 beta. Statistical analyses were measured using one-way analysis of variance for multimal comparisons. Data were presented as mean ± standard deviation (n = 6). aP < 0.001 vs control group; bP < 0.001, cP < 0.01, dP < 0.05 vs model group.

References 45.

1.	He Y, Koido M, Sutoh Y, et al. East Asian-specific and cross-ancestry genome-wide Meta-analyses provide mechanistic insights into peptic ulcer disease. Nat Genet 2023; 55: 2129-38.
2.	Xie X, Ren K, Zhou Z, Dang C, Zhang H. The global, regional and national burden of peptic ulcer disease from 1990 to 2019: a population-based study. BMC Gastroenterol 2022; 22: 58. DOI PMID
3.	Ford AC, Gurusamy KS, Delaney B, Forman D, Moayyedi P. Eradication therapy for peptic ulcer disease in Helicobacter pylori-positive people. Cochrane Database Syst Rev 2016; 4: CD003840.
4.	Li Z, Zou D, Ma X, et al. Epidemiology of peptic ulcer disease: endoscopic results of the systematic investigation of gastrointestinal disease in China. Am J Gastroenterol 2010; 105: 2570-7. DOI PMID
5.	Lanas A, Chan FKL. Peptic ulcer disease. Lancet 2017; 390: 613-24. DOI PMID
6.	Lau JY, Sung J, Hill C, Henderson C, Howden CW, Metz DC. Systematic review of the epidemiology of complicated peptic ulcer disease: incidence, recurrence, risk factors and mortality. Digestion 2011; 84: 102-13. DOI PMID
7.	Leow AH, Lim YY, Liew WC, Goh KL. Time trends in upper gastrointestinal diseases and Helicobacter pylori infection in a multiracial Asian population--a 20-year experience over three time periods. Aliment Pharmacol Ther 2016; 43: 831-7.
8.	Sverden E, Agreus L, Dunn JM, Lagergren J. Peptic ulcer disease. BMJ 2019; 367: l5495.
9.	Gupta A, Shetty S, Mutalik S, et al. Treatment of H. pylori infection and gastric ulcer: need for novel Pharmaceutical formulation. Heliyon 2023; 9: e20406.
10.	Kavitt RT, Lipowska AM, Anyane-Yeboa A, Gralnek IM. Diagnosis and treatment of peptic ulcer disease. Am J Med 2019; 132: 447-56. DOI PMID
11.	Haastrup PF, Thompson W, Sondergaard J, Jarbol DE. Side effects of long-term proton pump inhibitor use: a review. Basic Clin Pharmacol Toxicol 2018; 123: 114-21.
12.	Haenisch B, von Holt K, Wiese B, et al. Risk of dementia in elderly patients with the use of proton pump inhibitors. Eur Arch Psychiatry Clin Neurosci 2015; 265: 419-28.
13.	Kuna L, Jakab J, Smolic R, Raguz-Lucic N, Vcev A, Smolic M. Peptic ulcer disease: a brief review of conventional therapy and herbal treatment options. J Clin Med 2019; 8: 179.
14.	Gong AG, Li N, Lau KM, et al. Calycosin orchestrates the functions of Danggui Buxue Tang, a Chinese herbal decoction composing of Astragali Radix and Angelica Sinensis Radix: an evaluation by using calycosin-knock out herbal extract. J Ethnopharmacol 2015; 168: 150-7.
15.	Xia LF, Hao ZH, Wang J. Analysis of the effect of Danggui Buxue Jiawei decoction on the treatment of refractory gastric ulcer in the elderly. Zhong Guo She Qu Yi Shi 2018; 34: 99-100.
16.	Yang FX, Wang Y, Xia PF, et al. Review of chemical constituents,pharmacological effects and clinical applications of Danggui Buxue decoction and prediction and analysis of its Q-markers. Zhong Guo Zhong Yao Za Zhi 2021; 46: 2677-85.
17.	Li C, Zhu F, Wang S, Wang J, Wu B. Danggui Buxue decoction ameliorates inflammatory bowel disease by improving inflammation and rebuilding intestinal mucosal barrier. Evid Based Complement Alternat Med 2021; 2021: 8853141.
18.	Bi WP, Man HB, Man MQ. Efficacy and safety of herbal medicines in treating gastric ulcer: a review. World J Gastroenterol 2014; 20: 17020-8.
19.	Chen Z, Liu L, Gao C, et al. Astragali Radix (Huangqi): a promising edible immunomodulatory herbal medicine. J Ethnopharmacol 2020; 258: 112895.
20.	Kim A, Lim JW, Kim H, Kim H. Supplementation with Angelica keiskei inhibits expression of inflammatory mediators in the gastric mucosa of Helicobacter pylori-infected mice. Nutr Res 2016; 36: 488-97. DOI PMID
21.	Schottker B, Adamu MA, Weck MN, Brenner H. Helicobacter pylori infection is strongly associated with gastric and duodenal ulcers in a large prospective study. Clin Gastroenterol Hepatol 2012; 10: 487-93 e1.
22.	Lee YC, Dore MP, Graham DY. Diagnosis and treatment of Helicobacter pylori infection. Annu Rev Med 2022; 73: 183-95.
23.	Tarnawski AS, Ahluwalia A. The critical role of growth factors in gastric ulcer healing: the cellular and molecular mechanisms and potential clinical implications. Cells 2021; 10: 1964.
24.	Lian YZ, Lin IH, Yang YC, Chao JC. Gastroprotective effect of Lycium barbarum polysaccharides and C-phycocyanin in rats with ethanol-induced gastric ulcer. Int J Biol Macromol 2020; 165: 1519-28. DOI PMID
25.	Formiga RO, Alves Junior EB, Vasconcelos RC, et al. Effect of p-cymene and rosmarinic acid on gastric ulcer healing-involvement of multiple endogenous curative mechanisms. Phytomedicine 2021; 86: 153497.
26.	Hsieh HL, Yu MC, Cheng LC, et al. Quercetin exerts anti-inflammatory effects via inhibiting tumor necrosis factor-alpha-induced matrix metalloproteinase-9 expression in normal human gastric epithelial cells. World J Gastroenterol 2022; 28: 1139-58.
27.	Shams SGE, Eissa RG. Amelioration of ethanol-induced gastric ulcer in rats by quercetin: implication of Nrf2/HO1 and HMGB1/TLR4/NF-kappaB pathways. Heliyon 2022; 8: e11159.
28.	Fatima SF, Ishtiaq S, Lashkar MO, Youssef FS, Ashour ML, Elhady SS. Metabolic profiling of heliotropium crispum aerial parts using HPLC and FTIR and in vivo evaluation of its anti-ulcer activity using an ethanol induced acute gastric ulcer model. Metabolites 2022; 12: 750.
29.	Olaitan BS, Sabino DA, Pavan E, et al. Evidence for the involvement of cytokines modulation and prokinetic properties in gastric ulcer healing effects of helicteres sacarolha A. St.-Hil. A. Juss. Chem Biodivers 2022; 19: e202200322.
30.	Robinson K, Letley DP, Kaneko K. The human stomach in health and disease: infection strategies by Helicobacter pylori. Curr Top Microbiol Immunol 2017; 400: 1-26. DOI PMID
31.	Xiao ZP, Wang XD, Peng ZY, et al. Molecular docking, kinetics study, and structure-activity relationship analysis of quercetin and its analogous as Helicobacter pylori urease inhibitors. J Agric Food Chem 2012; 60: 10572-7.
32.	Yi L, Lu Y, Yu S, Cheng Q, Yi L. Formononetin inhibits inflammation and promotes gastric mucosal angiogenesis in gastric ulcer rats through regulating NF-kappa B signaling pathway. J Recept Signal Transduct Res 2022; 42: 16-22.
33.	Ding K, Tan YY, Ding Y, et al. beta-Sitosterol improves experimental colitis in mice with a target against pathogenic bacteria. J Cell Biochem 2019; 120: 5687-94.
34.	Zhang S, Shen Y, Liu H, et al. Inflammatory microenvironment in gastric premalignant lesions: implication and application. Front Immunol 2023; 14: 1297101.
35.	Sugimoto M, Yamaoka Y, Furuta T. Influence of interleukin polymorphisms on development of gastric cancer and peptic ulcer. World J Gastroenterol 2010; 16: 1188-200.
36.	Zou Y, Cui X, Xiang Q, et al. Protective effect of against ethanol-induced gastric ulcer and its mechanism. Zhejiang Da Xue Xue Bao Yi Xue Ban 2021; 50: 561-7.
37.	Arakawa T, Watanabe T, Tanigawa T, Tominaga K, Fujiwara Y, Morimoto K. Quality of ulcer healing in gastrointestinal tract: its pathophysiology and clinical relevance. World J Gastroenterol 2012; 18: 4811-22.
38.	Tourani M, Habibzadeh M, Karkhah A, Shokri-Shirvani J, Barari L, Nouri HR. Association of TNF-alpha but not IL-1beta levels with the presence of Helicobacter pylori infection increased the risk of peptic ulcer development. Cytokine 2018; 110: 232-6. DOI PMID
39.	Moezi L, Heidari R, Amirghofran Z, Nekooeian AA, Monabati A, Dehpour AR. Enhanced anti-ulcer effect of pioglitazone on gastric ulcers in cirrhotic rats: the role of nitric oxide and IL-1beta. Pharmacol Rep 2013; 65: 134-43.
40.	McKenna M, Balasuriya N, Zhong S, Li SS, O'Donoghue P. Phospho-form specific substrates of protein kinase B (AKT1). Front Bioeng Biotechnol 2020; 8: 619252.
41.	Siddika T, Balasuriya N, Frederick MI, Rozik P, Heinemann IU, O'Donoghue P. Delivery of active AKT1 to human cells. Cells 2022; 11: 3834.
42.	Tarnawski AS, Ahluwalia A, Jones MK. Angiogenesis in gastric mucosa: an important component of gastric erosion and ulcer healing and its impairment in aging. J Gastroenterol Hepatol 2014; 29 Suppl 4: 112-23.
43.	Ivyna de Araujo Rego R, Guedes Silvestre GF, Ferreira de Melo D, et al. Flavonoids-rich plant extracts against Helicobacter pylori infection as prevention to gastric cancer. Front Pharmacol 2022; 13: 951125.
44.	Zhang S, Huang J, Xie X, He Y, Mo F, Luo Z. Quercetin from polygonum capitatum protects against gastric inflammation and apoptosis associated with Helicobacter pylori infection by affecting the levels of p38MAPK, BCL-2 and BAX. Molecules 2017; 22: 744.
45.	Zheng Y, Liu Z, Cai A, et al. Study on the mechanism of Ginseng-Gegen for mesenteric lymphadenitis based on network pharmacology. Transl Pediatr 2022; 11: 1534-43. DOI PMID

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