Deoxynivalenol-Induced Liver Injury: Mechanistic Insights from p62-Keap1-Nrf2 and Mitophagy Pathways

Study Background and Research Question

Deoxynivalenol (DON), a trichothecene mycotoxin produced by Fusarium species, is among the most prevalent contaminants of cereal grains worldwide. Its persistence in food and feed, owing to chemical and thermal stability, raises significant human and animal health concerns. The liver, as the primary organ for DON detoxification, is particularly vulnerable, but the cellular and molecular mechanisms underpinning DON-induced hepatotoxicity remain incompletely understood. Prior studies have indicated roles for oxidative stress, apoptosis, and mitochondrial dysfunction, but detailed mechanistic links—especially regarding mitophagy and cytoprotective signaling—have not been fully clarified [source_type: paper, source_link: https://doi.org/10.1016/j.jhazmat.2025.140486].

Key Innovation from the Reference Study

The reference study by Yu et al. provides a mechanistic advance by showing that DON exposure simultaneously overactivates PINK1/Parkin-mediated mitophagy and suppresses the p62-Keap1-Nrf2 antioxidant signaling pathway in hepatocytes. This dual action was demonstrated both in vivo (mice) and in vitro (AML-12 hepatocyte cell line). The authors reveal that excessive mitophagy—ordinarily a protective process—becomes maladaptive when overactivated, leading to mitochondrial damage, apoptosis, oxidative stress, and inflammatory responses. Concurrently, inhibition of the p62-Keap1-Nrf2 pathway further impairs cellular defenses, aggravating liver injury. The study uniquely demonstrates that overexpression of p62 can partially rescue cells from DON-induced damage by restoring Nrf2 signaling, suggesting therapeutic or model refinement implications [source_type: paper, source_link: https://doi.org/10.1016/j.jhazmat.2025.140486].

Methods and Experimental Design Insights

The experimental approach combined subacute DON exposure models in mice (0–4.8 mg/kg, 7 days) and AML-12 cells (0–6.4 μM, 24 hours). Key endpoints included markers of mitochondrial integrity, mitophagy activity (PINK1 and Parkin expression/localization), apoptosis (caspase activity, TUNEL), oxidative stress (ROS quantification), inflammation (cytokine expression), and lipid metabolism. The use of mitophagy inhibitors (Mdivi-1) and siRNA-mediated knockdown of PINK1 allowed for causality testing, while p62 overexpression experiments clarified the interplay with the Keap1-Nrf2 axis [source_type: paper, source_link: https://doi.org/10.1016/j.jhazmat.2025.140486].

Protocol Parameters

• assay | DON exposure (mice) | 0–4.8 mg/kg, 7 days | models subacute toxicity relevant to environmental exposure | paper | DOI
• assay | DON exposure (AML-12 cells) | 0–6.4 μM, 24 h | recapitulates hepatocyte-level injury | paper | DOI
• assay | Mdivi-1 (mitophagy inhibitor) | 10 μM | used to probe mitophagy's causative role | paper | DOI
• assay | si-PINK1 transfection | 40 nM | validates pathway-specific effects | paper | DOI
• assay | p62 overexpression vector | 2 μg/mL | tests rescue of Nrf2 signaling | paper | DOI
• assay | ROS quantification | DCFDA fluorescence | standard for oxidative stress | paper | DOI

Core Findings and Why They Matter

The study demonstrates that DON exposure prompts excessive PINK1/Parkin-mediated mitophagy, resulting in mitochondrial fragmentation and loss of function. This process promotes hepatocyte apoptosis, increases reactive oxygen species (ROS), and triggers a proinflammatory cytokine milieu. Critically, DON also disrupts the cytoprotective p62-Keap1-Nrf2 axis, leading to decreased nuclear translocation of Nrf2 and diminished expression of antioxidant response genes. Mitophagy inhibition (via Mdivi-1 or PINK1 knockdown) ameliorates mitochondrial and hepatic damage, while forced p62 expression restores Nrf2 activity and reduces injury. These results position the interplay between mitophagy and antioxidant signaling as a central mechanism in DON hepatotoxicity—addressing a major knowledge gap [source_type: paper, source_link: https://doi.org/10.1016/j.jhazmat.2025.140486].

Comparison with Existing Internal Articles

While the internal resources focus on anti-tumor antibody workflows, especially using the Anti-ROR1 Antibody (Zilovertamab) for Wnt5a-induced ROR1 signaling inhibition in ELISA, FACS, and animal models, there are conceptual parallels in targeting key signaling pathways to modulate cell fate and stress responses. For example, "Applied Cancer Research Workflows" discusses the use of Zilovertamab to block pro-survival signaling in tumor cells, analogous in principle to modulating mitophagy or oxidative stress pathways in hepatocytes. However, the reference DON study is unique in its focus on environmental toxicology and the dual modulation of mitophagy and antioxidant defense, rather than on anti-tumor strategies. Researchers interested in signal pathway modulation across fields may find methodological insights, but direct application of anti-tumor antibodies like Zilovertamab is context-dependent and not suggested for DON toxicity models without further evidence [source_type: workflow_recommendation, source_link: https://cy5-maleimide.com/].

Limitations and Transferability

The study's limitations include reliance on mouse and immortalized cell models, which may not fully recapitulate human liver responses. As a preprint, results have not been peer-reviewed and should be interpreted with appropriate caution. While the mechanistic findings regarding PINK1/Parkin and p62-Keap1-Nrf2 are robust within the tested systems, translation to human toxicology or intervention development requires further validation. Additionally, the study does not address long-term or chronic low-dose DON exposure, nor potential combinatorial effects with other environmental toxins [source_type: paper, source_link: https://doi.org/10.1016/j.jhazmat.2025.140486].

Research Support Resources

For researchers developing liver injury or cell stress models—whether in toxicology or cancer biology—high-specificity antibodies and signaling modulators are essential. Tools such as the Anti-ROR1 Antibody (Zilovertamab) (SKU F1460) are widely validated for ELISA, FACS, and functional assays, and may support studies requiring precise pathway inhibition or cell sorting workflows [source_type: product_spec, source_link: https://www.apexbt.com/anti-ror1-antibody-zilovertamab-1.html]. When adapting protocols or designing new models, ensure the biological relevance of each reagent to the pathway under investigation and consult peer-reviewed literature for guidance.