Eicosapentaenoic Acid Modulates TGF-β1/Smad3/ILK Pathway to Attenuate Renal Fibrosis: A Biotechnological Approach

Zhiqiang Wei; Hao Ding; Haitao Li; Di Yin; Juan Cao

doi:10.30498/ijb.2025.513787.4098

Eicosapentaenoic Acid Modulates TGF-β1/Smad3/ILK Pathway to Attenuate Renal Fibrosis: A Biotechnological Approach

Iran J Biotechnol. 2025 Apr 1;23(2):e4098. doi: 10.30498/ijb.2025.513787.4098. eCollection 2025 Apr.

Authors

Zhiqiang Wei^{1

2}, Hao Ding^{1

2}, Haitao Li^{1

2}, Di Yin^{1

2}, Juan Cao^{1

2}

Affiliations

¹ Department of Nephrology, Taixing People's Hospital, Taixing, Jiangsu 225400, China.
² Taizhou Kidney Disease Clinical Research Center, Taixing, Jiangsu 225400, China.

Abstract

Background: Renal fibrosis is a key pathological process in chronic kidney disease (CKD), characterized by excessive extracellular matrix (ECM) deposition and epithelial-mesenchymal transition (EMT). Current treatment strategies have limited efficacy, necessitating the exploration of novel therapeutic agents. Eicosapentaenoic acid (EPA), a bioactive marine-derived omega-3 polyunsaturated fatty acid, has shown promise in modulating fibrosis-related signaling pathways.

Objectives: This study investigated the potential of EPA in mitigating renal fibrosis through the regulation of the transforming growth factor-β1 (TGF-β1)/Smad3/ILK pathway and its effects on ECM remodeling and EMT suppression in human kidney epithelial cells.

Materials and methods: Human Kidney-2 (HK-2) cells were subjected to albumin-induced EMT and treated with EPA, either alone or in combination with the β-catenin inhibitor LF3. The expression levels of key EMT markers (E-cadherin, N-cadherin, vimentin), ECM regulators (MMPs and TIMPs), and fibrosis-related signaling molecules (TGF-β1, Smad3, ILK) were assessed using immunofluorescence, ELISA, RT-qPCR, and Western blot analysis.

Results: EPA treatment significantly inhibited EMT by downregulating α-SMA, N-cadherin, vimentin, and active β-catenin while restoring E-cadherin expression (p < 0.05). ECM remodeling was evident through increased MMP-1, MMP-3, and MMP-9 expression and decreased TIMP-1 and TIMP-2 levels. Furthermore, EPA reduced TGF-β1, ILK, and phosphorylated Smad3 protein levels, an effect enhanced by LF3 co-treatment.

Conclusion: EPA shows preliminary potential as an antifibrotic agent in vitro by targeting the TGF-β1/Smad3/ILK pathway to regulate ECM remodeling and EMT suppression in renal fibrosis. This study provides insights into the EPA's application in medical biotechnology, particularly for CKD management.

Keywords: Eicosapentaenoic acid; Epithelial-mesenchymal transition; Extracellular matrix remodeling; Marine biotechnology; Renal fibrosis; TGF-β1/Smad3/ILK pathway.