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. 2024 Jul 19:15:1340309.
doi: 10.3389/fphar.2024.1340309. eCollection 2024.

Revealing the role of metformin in gastric intestinal metaplasia treatment

Ruoyu Hu #  1   2 , Xia Xue #  1 , Xiangdong Sun  1 , Yang Mi  1 , Huijuan Wen  1   2 , Huayuan Xi  1   2 , Fuhao Li  1   2 , Pengyuan Zheng  1   2 , Simeng Liu  1   2
Affiliations

Revealing the role of metformin in gastric intestinal metaplasia treatment

Ruoyu Hu et al. Front Pharmacol. .

Abstract

Objective: Gastric intestinal metaplasia (IM) is a precancerous stage associated with gastric cancer. Despite the observed beneficial effects of metformin on IM, its molecular mechanism remains not fully elucidated. This study aims to reveal the effects and potential mechanisms of metformin in treating IM based on both bioinformatics and in vivo investigations.

Methods: The seven public databases (GeneCards, DisGeNET, OMIM, SuperPred, Pharm Mapper, Swiss Target Prediction, TargetNet) were used in this work to identify targeted genes related to intestinal metaplasia (IM) and metformin. The shared targeted genes between metformin and IM were further analyzed by network pharmacology, while the interactions in-between were investigated by molecular docking. In parallel, the therapeutic effect of metformin was evaluated in IM mice model, while the core targets and pathways effected by metformin were verified in vivo.

Results: We screened out 1,751 IM-related genes and 318 metformin-targeted genes, 99 common genes identified in between were visualized by constructing the protein-protein interaction (PPI) network. The top ten core targeted genes were EGFR, MMP9, HIF1A, HSP90AA1, SIRT1, IL2, MAPK8, STAT1, PIK3CA, and ICAM1. The functional enrichment analysis confirmed that carcinogenesis and HIF-1 signaling pathways were primarily involved in the metformin treatment of IM. Based on molecular docking and dynamics, we found metformin affected the function of its targets by inhibiting receptor binding. Furthermore, metformin administration reduced the progression of IM lesions in Atp4a-/- mice model significantly. Notably, metformin enhanced the expression level of MUC5AC, while inhibited the expression level of CDX2. Our results also showed that metformin modulated the expression of core targets in vivo by reducing the activity of NF-κB and the PI3K/AKT/mTOR/HIF-1α signaling pathway.

Conclusion: This study confirms that metformin improves the efficacy of IM treatment by regulating a complex molecular network. Metformin plays a functional role in inhibiting inflammation/apoptosis-related pathways of further IM progression. Our work provides a molecular foundation for understanding metformin and other guanidine medicines in IM treatment.

Keywords: intestinal metaplasia; metformin; molecular docking; network pharmacology; precancerous diseases.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
The work flow of this study.
FIGURE 2
FIGURE 2
Pathway enrichment analyses of IM-related genes and metformin-targeted genes. (A) Top ten enriched KEGG pathways of IM-related genes. (B) Top ten enriched ReactomePA pathways of IM-related genes. (C) Top ten enriched KEGG pathways of metformin-targeted genes. (D) Top ten enriched ReactomePA pathways of metformin-targeted genes.
FIGURE 3
FIGURE 3
Common targets and PPI network of potential targets. (A) Venn diagram of 99 common targets. (B) The PPI network was constructed using the STRING and Cytoscape. (C) The top ten hub targets were selected from the PPI network.
FIGURE 4
FIGURE 4
GO enrichment analysis of targeted genes of metformin in IM treatment. The target genes are depicted by red circles, while brown circles represent the results of the enrichment analysis. The interconnecting lines between them refer to mutual relationships. The size of the circle indicates the level of the significance. (A) Biological processes of metformin in IM. (B) Cellular components of metformin in IM. (C) Molecular functions of metformin in IM.
FIGURE 5
FIGURE 5
KEGG enrichment analysis of targeted genes of metformin for IM treatment. The color represents the p-value, and the spot size represents the number of genes.
FIGURE 6
FIGURE 6
Docking patterns of metformin and its key target proteins. (A) Metformin- EGFR. (B) Metformin- MMP9. (C) Metformin- HIF1A. (D) Metformin- HSP90AA1. (E) Metformin- SIRT1. (F) Metformin- IL2. (G) Metformin- MAPK8. (H) Metformin- STAT1. (I) Metformin- PIK3CA. (J) Metformin- ICAM1.
FIGURE 7
FIGURE 7
Histopathological changes of gastric mucosa in mice. (A) H&E staining (n = 3). (B) AB-PAS staining (n = 3).
FIGURE 8
FIGURE 8
Metformin increases the level of MUC5AC and inhibits the expression of CDX2 in Atp4a−/− mice. (A) The expression of MUC5AC and CDX2 in the Atp4a−/− mice gastric tissues using IHC staining (n = 3). (B) The quantification of relative protein levels was performed using optical density analysis with ImageJ software. (*p < 0.05, **p < 0.01, ****p < 0.0001).
FIGURE 9
FIGURE 9
The mRNA expression of core targets and related factors. (A) The relative mRNA expression levels of MUC5AC and CDX2 (n = 5). (B) The heatmap of the relative mRNA expression levels of core target genes and pathway-related genes in three groups (n = 5). (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).
FIGURE 10
FIGURE 10
Metformin reduced the activation of NF-κB and inhibited the PI3K/AKT/mTOR/HIF-1α signaling pathway in the gastric tissues of Atp4a−/− mice. (A) Western blot was conducted to assess the protein levels of NF-κB, phospho-NF-κB, PI3K, phospho-PI3K, AKT, phospho-AKT, mTOR, phospho-mTOR, and HIF-1α (n = 3). (B–F) The relative protein levels were quantified (n = 3). (*p < 0.05, **p < 0.01).
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References

    1. Ala M., Ala M. (2021). Metformin for cardiovascular protection, inflammatory bowel disease, osteoporosis, periodontitis, polycystic ovarian syndrome, neurodegeneration, cancer, inflammation and senescence: what is next? ACS Pharmacol. Transl. Sci. 4, 1747–1770. 10.1021/acsptsci.1c00167 - DOI - PMC - PubMed
    1. Battista S., Ambrosio M. R., Limarzi F., Gallo G., Saragoni L. (2021). Molecular alterations in gastric preneoplastic lesions and early gastric cancer. Int. J. Mol. Sci. 22, 6652. 10.3390/ijms22136652 - DOI - PMC - PubMed
    1. Ba X., Huang Y., Shen P., Huang Y., Wang H., Han L., et al. (2021). WTD attenuating rheumatoid arthritis via suppressing angiogenesis and modulating the PI3K/AKT/mTOR/HIF-1α pathway. Front. Pharmacol. 12, 696802. 10.3389/fphar.2021.696802 - DOI - PMC - PubMed
    1. Berkman S. J., Roscoe E. M., Bourret J. C. (2019). Comparing self-directed methods for training staff to create graphs using Graphpad Prism. J. Appl. Behav. Anal. 52, 188–204. 10.1002/jaba.522 - DOI - PubMed
    1. Cheng J., Li C., Ying Y., Lv J., Qu X., Mcgowan E., et al. (2022). Metformin alleviates endometriosis and potentiates endometrial receptivity via decreasing VEGF and MMP9 and increasing leukemia inhibitor factor and HOXA10. Front. Pharmacol. 13, 750208. 10.3389/fphar.2022.750208 - DOI - PMC - PubMed

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