# Intramuscular iron supplementation enhances intestinal barrier function in weaned piglets challenged with enterotoxigenic Escherichia coli

**Authors:** Chenying Tian, Xiaofeng Zhang, Yecheng Xu, Linfeng Miao, Jing Zhao, Qingqing Xiong, Yuhui Zhang, Shouchuan Jiang, Yu Han, Huahua Du

PMC · DOI: 10.3389/fcimb.2025.1553639 · Frontiers in Cellular and Infection Microbiology · 2025-07-17

## TL;DR

Injecting iron into muscles helps piglets fight gut infections better than taking iron orally, reducing diarrhea and inflammation.

## Contribution

This study demonstrates that intramuscular iron supplementation is more effective than oral supplementation in mitigating ETEC-induced intestinal damage in piglets.

## Key findings

- Intramuscular iron reduced diarrhea rate by 53% and lowered pro-inflammatory cytokine levels.
- Intramuscular iron improved intestinal structure and tight junctions compared to oral iron.
- Intramuscular iron protected goblet cell function and enhanced iron metabolism in infected piglets.

## Abstract

Since iron is an essential mineral for both host and microbial communities, how to scientifically replenish the iron in the context of bacterial infection has become a critical issue. The aim of this study was to compare the influence of intramuscular and oral iron supplementation on the progression of bacterial infection.

Weaned piglets served as an experimental model for iron supplementation following enterotoxigenic Escherichia coli (ETEC) K88 infection. Piglets in control and oral iron supplementation groups received FeSO4 orally, while those in the intramuscular iron supplementation group were administered iron dextran (FeDex) via intramuscular injection. After challenge, piglets were euthanized, and serum and small intestinal tissues were collected for biochemical analysis, histological examination, inflammatory response assessment, gut microbiota profiling, and iron metabolism evaluation.

Intramuscular iron supplementation alleviated the clinical symptoms of bacterial infection, decreasing the diarrhea rate by 53% and mitigating the inflammatory response with lower serum levels of pro-inflammatory cytokines, such as TNF-α, IL-6, and IL-8. Compared to oral iron supplementation, intramuscular iron supplementation significantly mitigated the intestinal damage caused by ETEC K88 infection by increasing the ratio of villus length to crypt depth, and repairing epithelial tight junction. Furthermore, intramuscular iron supplementation also protected the function of intestinal goblet cells and improved iron metabolism of infected piglets.

Intramuscular iron supplementation is more effective during infection than oral iron supplementation.

## Linked entities

- **Chemicals:** FeSO4 (PubChem CID 24393), iron dextran (PubChem CID 105075), IL-6 (PubChem CID 165368475), IL-8 (PubChem CID 169410440)

## Full-text entities

- **Genes:** Il10 (interleukin 10) [NCBI Gene 16153] {aka CSIF, If2a, Il-10}, Tfrc (transferrin receptor) [NCBI Gene 22042] {aka 2610028K12Rik, CD71, E430033M20Rik, Mtvr1, TFR, TFR1}, Trf (transferrin) [NCBI Gene 22041] {aka Cd176, HP, Tf, Tfn, hpx}, Slc40a1 (solute carrier family 40 (iron-regulated transporter), member 1) [NCBI Gene 53945] {aka Dusg, Fpn1, IREG1, MTP, MTP1, Ol5}, Aoc1 (amine oxidase, copper-containing 1) [NCBI Gene 76507] {aka 1600012D06Rik, Abp1, DAO}, Ocln (occludin) [NCBI Gene 18260] {aka Ocl}, Il1b (interleukin 1 beta) [NCBI Gene 16176] {aka IL-1beta, Il-1b}, TNF (tumor necrosis factor) [NCBI Gene 397086] {aka TNFSF2, TNFa}, Dao (D-amino acid oxidase) [NCBI Gene 13142] {aka DAAO, DAMOX, Dao-1, Dao1}, Cldn1 (claudin 1) [NCBI Gene 12737], Epas1 (endothelial PAS domain protein 1) [NCBI Gene 13819] {aka HIF-2alpha, HIF2A, HLF, HRF, MOP2, bHLHe73}, Igha (immunoglobulin heavy constant alpha) [NCBI Gene 238447] {aka IgA, Igh-2}, Fth1 (ferritin heavy polypeptide 1) [NCBI Gene 14319] {aka FHC, Fth, HFt, MFH}, IL1B (interleukin 1 beta) [NCBI Gene 3553] {aka IL-1, IL1-BETA, IL1F2, IL1beta}, Slc11a2 (solute carrier family 11 (proton-coupled divalent metal ion transporters), member 2) [NCBI Gene 18174] {aka DCT1, DMT1, Nramp2, mk, van}, ZO-1 [NCBI Gene 396567], IL6 (interleukin 6) [NCBI Gene 3569] {aka BSF-2, BSF2, CDF, HGF, HSF, IFN-beta-2}, CD4 (CD4 molecule) [NCBI Gene 920] {aka CD4mut, IMD79, Leu-3, OKT4D, T4}, Actb (actin, beta) [NCBI Gene 11461] {aka Actx, E430023M04Rik, beta-actin}, CD4 (CD4 molecule) [NCBI Gene 407098], ALB (albumin) [NCBI Gene 280717], Il4 (interleukin 4) [NCBI Gene 16189] {aka BSF-1, Il-4}, TNF (tumor necrosis factor) [NCBI Gene 7124] {aka DIF, IMD127, TNF-alpha, TNFA, TNFSF2, TNLG1F}, Ifng (interferon gamma) [NCBI Gene 15978] {aka IFN-g, If2f, Ifg}, Nos2 (nitric oxide synthase 2, inducible) [NCBI Gene 18126] {aka MAC-NOS, NOS-II, Nos-2, Nos2a, i-NOS, iNOS}, Cxcl15 (C-X-C motif chemokine ligand 15) [NCBI Gene 20309] {aka Il8, Scyb15, lungkine, weche}, OCLN (occludin) [NCBI Gene 397236], Cd4 (CD4 antigen) [NCBI Gene 12504] {aka L3T4, Ly-4}, Tnf (tumor necrosis factor) [NCBI Gene 21926] {aka DIF, TNF-a, TNF-alpha, TNFSF2, TNFalpha, Tnfa}
- **Diseases:** diarrhea (MESH:D003967), bacterial (MESH:D001424), iron (MESH:D000090463), villi injury (MESH:D014947), atrophy (MESH:D001284), Intestinal infections (MESH:D007410), chronic (MESH:D002908), lethargy (MESH:D053609), IBD (MESH:D015212), iron deficiency anemia (MESH:D018798), death (MESH:D003643), systemic (MESH:D015619), infected (MESH:D007239), colonic inflammation (MESH:D007249), iron metabolism disorders (MESH:D019189), vomiting (MESH:D014839)
- **Chemicals:** Paraffin (MESH:D010232), ferrous sulfate (MESH:C020748), Trizol (MESH:C411644), hematoxylin (MESH:D006416), SYBR Green (MESH:C098022), phosphate (MESH:D010710), glutaraldehyde (MESH:D005976), AB (MESH:D000423), luminal (MESH:D010634), ethanol (MESH:D000431), acetone (MESH:D000096), isoamyl acetate (MESH:C020377), ETEC K88 (-), Spurr resin (MESH:C048709), oxygen (MESH:D010100), palladium (MESH:D010165), Fe (MESH:D007501), uranyl acetate (MESH:C005460), EDTA (MESH:D004492), paraformaldehyde (MESH:C003043), eosin (MESH:D004801), ROS (MESH:D017382), H&amp;E (MESH:D006371), LPS (MESH:D008070), osmic acid (MESH:D009993), FeDex (MESH:C011819), water (MESH:D014867), D-LA (MESH:D019344), Iron dextran (MESH:D007505)
- **Species:** Escherichia coli (E. coli, species) [taxon 562], Escherichia coli O157:H7 (no rank) [taxon 83334], Sus scrofa (pig, species) [taxon 9823], Homo sapiens (human, species) [taxon 9606], Lactobacillus (genus) [taxon 1578], Mus musculus (house mouse, species) [taxon 10090]
- **Cell lines:** ETEC K88 — Homo sapiens (Human), Burkitt lymphoma, Cancer cell line (CVCL_IZ97), ETEC — Mus musculus (Mouse), Hybridoma (CVCL_C5CN)

## Full text

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## Figures

8 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12310581/full.md

## References

36 references — full list in the complete paper: https://tomesphere.com/paper/PMC12310581/full.md

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Source: https://tomesphere.com/paper/PMC12310581