Open Access

ARTICLE

YOD1 Stabilizes RIPK1 via Deubiquitination to Activate NF-κB and Promote Cardiomyocyte H/R Injury

Liangliang Liu, Linjun Wang, Xin Song, Zhen Liu^*

Department of Cardiology, Lianyungang Hospital of Traditional Chinese Medicine, Lianyungang, China

* Corresponding Author: Zhen Liu. Email:

BIOCELL 2026, 50(9), 10 https://doi.org/10.32604/biocell.2026.080414

Received 09 February 2026; Accepted 13 May 2026; Issue published 26 August 2026

Abstract

Background: Myocardial ischemia-reperfusion (I/R) injury represents a severe pathological process in cardiovascular diseases. This study aims to elucidate the mechanism of the yeast ovarian tumor (OTU) domain-containing protein 1 (YOD1) in cardiomyocyte injury. Methods: A hypoxia/reoxygenation (H/R) model was established using human AC16 cells to simulate I/R injury in vitro. Reverse transcription quantitative PCR (RT-qPCR) and western blotting were used to detect gene and protein expression. Cellular functions were evaluated using Cell Counting Kit-8 (CCK-8), lactate dehydrogenase (LDH), enzyme-linked immunosorbent assay (ELISA), flow cytometry, and biochemical kits. Protein interaction was validated through co-immunoprecipitation (Co-IP), ubiquitination assays, and cycloheximide (CHX) chase experiments. Results: YOD1 was up-regulated in H/R-induced AC16 cells (p < 0.01). H/R decreased cell viability (p < 0.001), superoxide dismutase (SOD) (p < 0.001), and glutathione peroxidase 4 (GPX4) (p < 0.001) and increased LDH (p < 0.001), interleukin-6 (IL-6) (p < 0.001), tumor necrosis factor-alpha (TNF-α) (p < 0.001), reactive oxygen species (ROS) (p < 0.001), malondialdehyde (MDA) (p < 0.001), and Fe²⁺ (p < 0.001); these effects were alleviated by YOD1 knockdown (p < 0.01). YOD1 knockdown (p < 0.001) also reversed H/R-increased receptor-interacting serine/threonine kinase 1 (RIPK1) protein levels (p < 0.001) and p-p65/p65 (p < 0.001) and p-nuclear factor-kappa-B inhibitor alpha (p-IκBα)/IκBα (p < 0.001) ratios. RIPK1 overexpression reversed the protective effects of YOD1 knockdown via activating the nuclear factor-kappa-B (NF-κB) pathway (p < 0.01). Mechanistically, YOD1 interacted with RIPK1 and inhibited its K63-linked ubiquitination degradation (p < 0.01). These statistical significances support the clinical potential of targeting YOD1 to mitigate inflammation, oxidative stress, and ferroptosis-related changes in myocardial I/R injury. Conclusion: YOD1 stabilizes RIPK1 via K63-linked ubiquitination to activate the NF-κB pathway, exacerbating inflammation, oxidative stress, and ferroptosis-related changes in H/R-induced cardiomyocytes. YOD1 may represent a potential therapeutic target for myocardial I/R injury, though further in vivo and clinical validation is required.

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Keywords

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Supplementary Material

Supplementary Material File

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