Abstract: TH-PO0162
Study on the Mechanism of PRDX5 Regulating ACSL4 Ubiquitination via UFD1 in Ferroptosis Associated with Contrast-Induced Nephropathy
Session Information
- AKI: Mechanisms - 1
November 06, 2025 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Acute Kidney Injury
- 103 AKI: Mechanisms
Authors
- Zhou, Fangfang, Ningbo No 2 Hospital, Ningbo, Zhejiang, China
- Xu, Youjun, Ningbo No 2 Hospital, Ningbo, Zhejiang, China
- Guo, Zhenyu, Ningbo No 2 Hospital, Ningbo, Zhejiang, China
- Wang, Lailiang, Ningbo No 2 Hospital, Ningbo, Zhejiang, China
- Zheng, Xingyue, Ningbo No 2 Hospital, Ningbo, Zhejiang, China
- Luo, Qun, Ningbo No 2 Hospital, Ningbo, Zhejiang, China
Background
Contrast-induced nephropathy (CIN) is a common complication caused by iodinated contrast media, with no specific treatment available. Ferroptosis may contribute to its pathogenesis, and peroxiredoxin 5 (PRDX5) is downregulated in the urine of CIN patients. This study investigates whether PRDX5 affects ferroptosis in CIN by regulating the ubiquitination of acyl-CoA synthetase long-chain family member 4 (ACSL4).
Methods
HK-2 cells and a mouse CIN model (induced by iohexol) were established. Groups included: control, CIN, CIN + ferroptosis inhibitor (Fer-1), PRDX5 overexpression, and PRDX5 overexpression + CIN. PRDX5, ACSL4, GPX4, and SLC7A11 expression were analyzed (qPCR/Western blot). Cell viability was measured via MTT assay. Fe2+and reactive oxygen species (ROS) levels were quantified. Co-immunoprecipitation (Co-IP) combined with LC-MS/MS identified UFD1 as a PRDX5-interacting protein. Immunofluorescence confirmed co-localization. Ubiquitination assays evaluated UFD1-mediated regulation of ACSL4. Renal function and histopathology were assessed in mice.
Results
1.Ferroptosis in CIN: The CIN group exhibited reduced cell proliferation, increased ACSL4, decreased GPX4/SLC7A11, and elevated Fe2+/ROS levels, which were reversed by Fer-1 (P<0.05).
2.Protective role of PRDX5: PRDX5 expression was downregulated in CIN. Its overexpression alleviated cellular damage, suppressed ACSL4, restored GPX4/SLC7A11 levels, and reduced Fe2+/ROS accumulation. In mice, PRDX5 overexpression improved renal injury and lowered serum creatinine.
3.PRDX5-UFD1 interaction: UFD1 was identified as a PRDX5-binding partner via mass spectrometry. Co-IP and immunofluorescence confirmed their co-localization, which weakened in CIN.
4.UFD1 regulates ACSL4 ubiquitination: UFD1 knockdown reduced ACSL4 ubiquitination and degradation, leading to its accumulation. UFD1 overexpression or proteasome inhibitor MG132 reversed this effect. Immunofluorescence demonstrated co-localization of UFD1 with ACSL4 and ubiquitin, suggesting UFD1 binds ACSL4’s intracellular domain.
Conclusion
PRDX5 inhibits ferroptosis and mitigates CIN by binding UFD1 to promote ACSL4 ubiquitination and degradation. These findings propose novel therapeutic strategies targeting the PRDX5-UFD1-ACSL4 axis in CIN.
Funding
- Government Support – Non-U.S.