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Updated: May 4, 2026
Induction of Drug-Induced, Autoimmune Hepatitis in BALB/c Mice for the Study of Its Pathogenic Mechanisms
Published on: May 29, 2020
An intrabody targeting PHD2 protects mice against acetaminophen-induced liver injury by attenuating ferroptosis
Ze Ren1, Ying Wang1, Cheng Cheng1
1Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, School of Life Sciences, Jilin University, Changchun, 130012, China.
Abstract:
Acetaminophen (APAP) overdose has become the most common cause of acute liver failure. Ferroptosis plays a critical role in APAP-induced liver injury (AILI). Although inhibiting prolyl hydroxylase 2 (PHD2)-mediated degradation of hypoxia-inducible factor-1α (HIF-1α) can alleviate APAP hepatotoxicity, the function and underlying mechanism of PHD2/HIF-1 axis in APAP-induced ferroptosis are not fully understood. Here, we developed a novel anti-PHD2 cytoplasmic intrabody (INP2) and elucidated the anti-ferroptosis mechanism by which INP2 inhibits APAP hepatotoxicity. Our findings demonstrated that INP2 specifically recognized PHD2 and disrupted its interaction with HIF-1α, inhibiting HIF-1α hydroxylation and increasing HIF-1α stability. In the murine AILI model, INP2 pretreatment significantly protects against AILI by attenuating ferroptosis via restoring redox homeostasis, attenuating malondialdehyde and ROS accumulation, and decreasing ferrous iron content. Importantly, INP2 enhanced glutathione biosynthesis and eliminated oxidative stress by upregulating SLC7A11 and glutathione peroxidase 1 (GPX1), thereby decreasing cytotoxicity in APAP/H2O2-exposed hepatocytes. GPX1 knockdown markedly impaired INP2-mediated hepatoprotection and anti-ferroptosis by suppressing glutathione-dependent reduction of cytotoxic lipid hydroperoxides including 12-HPETE. Collectively, a PHD2-targeted intrabody protects acetaminophen-induced liver injury by attenuating ferroptosis via regulating PHD2/HIF-1α/GPX1 axis, which enhances glutathione biosynthesis, suppresses oxidative stress and ferroptosis, offering a potential strategy for the prevention and treatment of AILI.
Insights
A novel intrabody targeting prolyl hydroxylase 2 (PHD2) protects against acetaminophen-induced liver injury by inhibiting ferroptosis. This approach restores redox balance and enhances glutathione synthesis, offering a new therapeutic strategy.
Area of Science:
- Biochemistry
- Hepatology
- Molecular Biology
Background:
- Acetaminophen overdose is a leading cause of acute liver failure, with ferroptosis playing a key role.
- The prolyl hydroxylase 2 (PHD2)/hypoxia-inducible factor-1α (HIF-1α) axis is implicated in acetaminophen-induced liver injury (AILI), but its role in ferroptosis is unclear.
Purpose of the Study:
- To develop a novel anti-PHD2 intrabody (INP2) and investigate its mechanism in preventing AILI by targeting ferroptosis.
- To elucidate how INP2 regulates the PHD2/HIF-1α axis and its downstream effects on ferroptosis and hepatoprotection.
Main Methods:
- Development of a cytoplasmic intrabody (INP2) targeting PHD2.
- Assessment of INP2's effect on PHD2/HIF-1α interaction, HIF-1α stability, and ferroptosis markers in vitro and in a murine AILI model.
- Investigation of INP2's impact on redox homeostasis, glutathione biosynthesis, and the expression of SLC7A11 and glutathione peroxidase 1 (GPX1).
Main Results:
- INP2 specifically recognized PHD2, disrupted its interaction with HIF-1α, and increased HIF-1α stability.
- INP2 pretreatment significantly protected against AILI by attenuating ferroptosis, restoring redox homeostasis, and reducing oxidative stress markers.
- INP2 upregulated SLC7A11 and GPX1, enhancing glutathione biosynthesis and decreasing hepatocyte cytotoxicity; GPX1 knockdown impaired INP2's protective effects.
Conclusions:
- A PHD2-targeted intrabody (INP2) effectively protects against acetaminophen-induced liver injury by attenuating ferroptosis.
- The mechanism involves regulating the PHD2/HIF-1α/GPX1 axis, which enhances glutathione synthesis and suppresses oxidative stress.
- INP2 represents a promising therapeutic strategy for preventing and treating acetaminophen-induced liver injury.
