Wei Zhiqiang, Ding Hao, Li Haitao, Yin Di, Cao Juan
Department of Nephrology, Taixing People's Hospital, Taixing, Jiangsu 225400, China.
Taizhou Kidney Disease Clinical Research Center, Taixing, Jiangsu 225400, China.
Iran J Biotechnol. 2025 Apr 1;23(2):e4098. doi: 10.30498/ijb.2025.513787.4098. eCollection 2025 Apr.
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.
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.
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.
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.
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.
肾纤维化是慢性肾脏病（CKD）的关键病理过程，其特征为细胞外基质（ECM）过度沉积和上皮-间质转化（EMT）。目前的治疗策略疗效有限，因此需要探索新型治疗药物。二十碳五烯酸（EPA）是一种具有生物活性的海洋来源ω-3多不饱和脂肪酸，已显示出在调节纤维化相关信号通路方面的潜力。
本研究通过调节转化生长因子-β1（TGF-β1）/Smad3/整合素连接激酶（ILK）通路，研究EPA减轻肾纤维化的潜力及其对人肾上皮细胞中ECM重塑和EMT抑制的影响。
人肾-2（HK-2）细胞接受白蛋白诱导的EMT，并单独或与β-连环蛋白抑制剂LF3联合用EPA处理。使用免疫荧光、酶联免疫吸附测定（ELISA）、逆转录定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹分析评估关键EMT标志物（E-钙黏蛋白、N-钙黏蛋白、波形蛋白）、ECM调节因子（基质金属蛋白酶（MMPs）和基质金属蛋白酶组织抑制因子（TIMPs））以及纤维化相关信号分子（TGF-β1、Smad3、ILK）的表达水平。
EPA处理通过下调α-平滑肌肌动蛋白（α-SMA）、N-钙黏蛋白、波形蛋白和活性β-连环蛋白，同时恢复E-钙黏蛋白表达，显著抑制EMT（p<0.05）。通过增加MMP-1、MMP-3和MMP-9表达以及降低TIMP-1和TIMP-2水平，ECM重塑明显。此外，EPA降低了TGF-β1、ILK和磷酸化Smad3蛋白水平，LF3联合处理增强了这一效果。
EPA通过靶向TGF-β1/Smad3/ILK通路调节肾纤维化中的ECM重塑和EMT抑制，在体外显示出作为抗纤维化药物的初步潜力。本研究为EPA在医学生物技术中的应用提供了见解，特别是在CKD管理方面。
