RARE DISEASE
Paracetamol poisoning
Paracetamol poisoning
Paracetamol poisoning
Synonyms: Acetaminophen poisoning
Synonyms: Acetaminophen poisoning
Synonyms: Acetaminophen poisoning
Drug discovery
6
drugs
With orphan designations
Overview
Paracetamol (acetaminophen) poisoning occurs when excessive doses overwhelm hepatic glutathione reserves, leading to toxic metabolite (NAPQI)-induced hepatotoxicity. Initial symptoms (nausea, vomiting, abdominal pain) may progress to liver failure, coagulopathy, and multi-organ dysfunction within 72 hours. Diagnosis relies on timed serum paracetamol levels plotted on the Rumack-Matthew nomogram [1][7]. First-line treatment with intravenous N-acetylcysteine (NAC) within 8-10 hours of ingestion prevents liver damage by replenishing glutathione [3][9][14].
Population
Common in intentional self-harm among adolescents/young adults in high-income countries [9][16], with 25,000 annual UK hospitalizations [9].
Accidental pediatric ingestions account for 6% of poisonings globally [8][13], particularly in children <7 years [10].
High-risk groups: chronic alcohol users, malnourished patients, and those taking enzyme-inducing medications (e.g., anticonvulsants) [1][5].
Burden
Leading cause of acute liver failure in the US/UK, contributing to 56% of severe hepatic injury cases [8][13].
Mortality: 0.1–0.4% in treated cases, rising to 95% in untreated patients with acidosis (pH <7.3) [4][11].
Economic impact: 492 deaths (1996–2019) and 5,666 hospitalizations in Brazil alone [2]; liver transplants cost >$500,000 USD per case [13][17].
Therapies
Immediate decontamination: Activated charcoal within 1-2 hours for ingestions >75–100 mg/kg [3][7].
Antidote: Weight-based NAC infusion (two-bag regimen: 200 mg/kg over 4h → 100 mg/kg over 16h) [7][14], reducing adverse reactions vs older protocols.
Risk stratification: NAC initiated if serum paracetamol exceeds nomogram thresholds (≥100 mg/L at 4h) or for staggered/uncertain-time overdoses [1][7][14].
Categories: rare disorders due to toxic effects
Research Papers
2,668 drug discovery papers about Paracetamol poisoning, with 16 next-in-class emerging drug candidates forecasted to outperform the average preclinical success rate. Recent publications:
2,668 drug discovery papers about Paracetamol poisoning, with 16 next-in-class emerging drug candidates forecasted to outperform the average preclinical success rate. Recent publications:
categories:
Small molecules
small molecules
2026-08-17 | Subchronic Alcohol Exposure Exacerbates Acetaminophen-Induced Acute Liver Injury via Macrophage-Mediated Inflammation and Mitochondrial Dysfunction.
Alcohol use is a major coexposure in patients taking acetaminophen (APAP), yet how subchronic alcohol exposure reshapes hepatic vulnerability to APAP remains incompletely defined. In this study, we examined the effects of chronic alcohol intake on APAP-induced liver injury in mice. Our results indicated that subchronic alcohol exposure significantly increased the mortality rate in APAP-overdosed mice. Alcohol pretreatment also exacerbated APAP-induced hepatic damage, as evidenced by elevated serum ALT and AST levels, enhanced inflammatory responses, and aggravated oxidative stress. Quantitative polymerase chain reaction (qPCR) analysis revealed that alcohol consumption suppressed mitochondrial biogenesis (MB)-related genes, including Pgc-1α and transcription factor A, mitochondrial (TFAM). Notably, macrophage depletion via clodronate liposomes not only attenuated hepatocyte injury and inflammation induction but also restored the expression of MB/function-related genes, including Pgc-1α, TFAM, and Opa1. These findings suggest macrophage-involved inflammatory amplification may aggravate APAP-induced liver injury, at least in part, by impairing mitochondrial adaptive repair responses.
2026-08-11 | Clinical Characteristics and Pattern of Paracetamol-Induced Liver Injury at Hospital Shah Alam: A Three-Year Retrospective Case Series
Background: Paracetamol poisoning is the most common type of therapeutic drug poisoning seen globally. The dose of ingested paracetamol and the time of presentation to a healthcare facility are important factors in assessing the severity of poisoning. In addition, factors affecting hepatic glutathione levels further influence the risk of toxicity. Objective: This three years case series aims to analyse the characteristics of liver injury observed in patients who presented to the hospital’s ED for Shah Alam following an intentional paracetamol overdose. Additionally, it seeks to determine risk factors specific to this local population. Methodology: Patients with a history of intentional paracetamol overdose and biochemical evidence of liver injury who presented to the ED of Hospital Shah Alam between 1st January 2022 and 31st December 2024 were included in this study. Patients aged less than 18 years were excluded from this study. Results: A total of 13 patients were included in this case series. The pattern of liver injury followed an established clinical course; however, no patients developed liver failure. High doses of paracetamol ingestion and late presentation after overdose were the expected risk factors. A further risk factor identified in this case series was low body weight. Six patients in this case series weighed less than 45 kg. Underweight patients may have underlying malnutrition. Paracetamol overdose hepatotoxicity risk is linked to depleted hepatic glutathione, which is known to be due to malnutrition. Conclusion: High doses of paracetamol ingestion with late presentation to a healthcare facility are known risk factors for paracetamol-induced liver injury. This case series also revealed that underweight patients are at risk, possibly because of malnutrition. These findings suggest that clinicians may consider a lower treatment threshold, such as the 100-line threshold, in underweight or potentially malnourished patients, although larger studies are needed.
2026-08-11 | Defining risk: the case for conceptual clarity in acetaminophen poisoning
Despite the widespread availability of acetylcysteine, acetaminophen (paracetamol) poisoning remains the most common cause of acute liver failure and acute liver transplantation in the United State...
2026-08-07 | Acetaminophen Poisoning: From Molecular Mechanisms to Multidisciplinary Management - A Contemporary Clinical Overview
Acetaminophen (N-acetyl-para-aminophenol, paracetamol) remains the leading cause of acute liver failure in the United States and accounts for a substantial proportion of pharmaceutical overdoses globally. This comprehensive review examines the epidemiology, pathophysiology, clinical presentation, and evidence-based management of acetaminophen toxicity, with particular emphasis on the critical role of timely intervention. The drug's widespread availability and pervasive public perception of safety contribute to its prominence in toxicity statistics, with over 60 million Americans using acetaminophen weekly, often unknowingly through combination products. The pathophysiological cascade is initiated by the saturation of hepatic sulfation and glucuronidation pathways, leading to the formation of the highly reactive metabolite N-acetyl-p-benzoquinoneimine (NAPQI) through cytochrome P450-mediated oxidation, predominantly via CYP2E1. When hepatic glutathione stores are depleted, NAPQI binds covalently to mitochondrial proteins, triggering oxidative stress, lipid peroxidation, and ultimately centrilobular necrosis. The clinical course classically progresses through four distinct stages, with the Rumack-Matthew nomogram serving as the cornerstone for risk stratification following acute ingestion. N-acetylcysteine, the specific antidote, is most effective when administered within 8 hours of ingestion and acts through glutathione replenishment, direct conjugation with NAPQI, and anti-inflammatory mechanisms. Emerging evidence suggests a potential role for fomepizole as adjunctive therapy in high-risk patients, though this remains investigational. Prognosis is excellent with early treatment, but delays significantly increase morbidity and mortality, with approximately 1-3% of severe cases requiring liver transplantation. A multidisciplinary approach encompassing emergency medicine, toxicology, hepatology, and critical care is essential for optimal patient outcomes. This review underscores the imperative for enhanced patient education, improved labeling practices, and continued research into novel therapeutic strategies to further reduce the burden of acetaminophen-induced hepatotoxicity.
2026-07-31 | Boosting Thioredoxin-1 Protects Against Acetaminophen-Induced Cardiotoxicity
Background: Although overdose-induced acute hepatic failure following acetaminophen (APAP) administration is well documented, clinical studies have also reported myocardial infarction, cardiac dysfunction, arrhythmias, pericarditis, and cardiac myocyte necrosis in APAP poisoning. As APAP overdose is associated with excessive reactive oxygen species (ROS) production and metabolism-mediated glutathione depletion, the cytosolic thioredoxin system (Trx-1/TrxR1/NADPH) may offer an alternative mechanism for cardioprotection. Although transgenic mice overexpressing thioredoxin-1 (Trx-1) are protected against doxorubicin/Adriamycin-induced cardiotoxicity, whether pharmacological enhancement of the cytosolic Trx system can mitigate APAP-induced cardiotoxicity and improve the therapeutic efficacy of chlorpromazine (CPZ) remains unclear. Methods: Cardiotoxicity was induced in wild-type C57BL/6 mice using APAP (400 mg/kg, i.p.). Expression of the Trx system was upregulated via chronic daily administration of low-dose epinephrine (EP, 0.2 mg/kg, s.c. for 14 days), and mice were subsequently treated with a subtoxic dose of CPZ (6 mg/kg, i.p.). Results: EP significantly upregulated the expression of Trx-1, thioredoxin-like protein 1 (Trx-L1), and thioredoxin reductase 1 (TrxR1) in cardiac tissue. Furthermore, EP attenuated APAP-induced acute cardiac injury, and its combination with CPZ additively amplified these cardioprotective effects. These findings were supported by the normalization of serum cardiac biomarkers (aspartate aminotransferase [AST], lactate dehydrogenase [LDH], and cardiac troponin I [cTnI]), reduction of oxidative stress (restoration of reduced glutathione [GSH], superoxide dismutase [SOD], and nuclear factor erythroid 2-related factor 2 [Nrf2]) and inflammatory markers (tumor necrosis factor-alpha [TNF-α]) in cardiac tissue, and preservation of normal myocardial histological architecture. Mechanistically, activation of the Trx system was accompanied by upregulation of β-arrestin-1 and cAMP response element-binding protein 1 (CREB1), indicating their involvement in Trx-mediated cardioprotection. Conclusion: Pharmacological enhancement of the cytosolic thioredoxin system protects against APAP-induced acute cardiac injury, and combining EP with CPZ exerts an additive cardioprotective effect.
cell therapies
2026-07-21 | Adoptive transfer of VSIG4+ macrophages interrupts the CCL2-CCR2 inflammatory amplification loop to mitigate acetaminophen-induced acute liver injury in murine and human organoid models.
Acute liver injury (ALI) can rapidly progress to life-threatening acute liver failure. Liver transplantation remains the only definitive treatment, despite critical donor shortages. Macrophage-based therapies have shown promise in ALI but face challenges related to subset imprecision and limited humanized validation. VSIG4 is generally considered a marker of Kupffer cells; however, it is also expressed in monocyte-derived macrophages (MoMFs), and its role remains unclear during ALI. By integrating single-cell RNA sequencing (scRNA-seq) data from human and mouse liver tissues with clinical ALI tissue samples, we elucidated the dynamic changes in VSIG4+ macrophages (VSIG4+ Mφ) within the liver. We developed a reversible immunomagnetic nanoparticle system for the non-destructive isolation of viable VSIG4+ Mφ. The therapeutic efficacy and potential mechanisms of VSIG4+ Mφ were evaluated through tissue and molecular-level analyses in an acetaminophen (APAP)-induced ALI mouse model, as well as in a newly established vascularized human liver organoid ALI and monocyte chemotaxis model. scRNA-seq revealed a previously underrecognized subpopulation of VSIG4+ MoMFs, which increases following ALI, while the number of resident VSIG4+ Kupffer cells decreases significantly. Clinical ALI tissue samples also confirmed the presence of CCR2+VSIG4+ cells in the livers of patients with ALI. In the APAP-induced ALI animal model, adoptive transfer of isolated VSIG4+ MoMFs dramatically decreased serum alanine transaminase, hepatic necrosis, and apoptosis while increasing anti-inflammatory cytokines. In contrast, adoptive transfer of unselected BMDM failed to improve liver injury and instead exacerbated certain pro-inflammatory responses, including elevated TNF-α and reduced CD206 expression. Mechanistically, VSIG4+ MoMFs prevented inflammatory amplification by suppressing NF-κB-dependent CCL2 transcription, thereby disrupting the CCL2-CCR2 chemotactic axis and reducing pro-inflammatory CCR2+ monocyte and macrophage recruitment. We further developed vascularized human liver organoids and APAP-induced hepatocyte injury and monocyte chemotaxis, finding that the chemotaxis-interrupting mechanism was fully recapitulated in the human liver organoid ALI model. This study identifies VSIG4+ MoMFs as a therapeutically viable subset for ALI, with clear superiority over unselected BMDM. By blocking the CCL2-CCR2 inflammatory amplification loop, these cells attenuate liver injury in both mouse and humanized models. These consistent findings provide robust preclinical evidence to support the advancement of VSIG4+ Mφ-based immunotherapies into clinical practice.
2026-03-18 | Paracetamol Poisoning and Liver Transplantation: An Updated Review
Introduction:: Regarding clinical recovery and survival of patients with severe paracetamol- induced acute liver failure and hepatic encephalopathy, liver transplantation is recommended as a good approach to management. This review summarizes the findings of previous studies associated with paracetamol-induced hepatotoxicity and liver transplantation. Methods:: All relevant articles (n=113) were searched systematically between the relevant record initiation and 14 October 2025 (the date of our final search), across PubMed and Web of Science. The search terms included the main concepts and their synonyms, including the problem: “paracetamol- over-dose”, “paracetamol poisoning”, the intervention “assessment of hepatotoxicity”, and the outcome “liver transplantation”, combined with the AND, OR operators. Synonyms for this concept were extracted from thesauri such as Medical Subject Headings (MeSH). Results:: An arterial pH lower than 7.3 or all three of the following: 1) International Normalized ratio of Prothrombin Time > 6.5, 2) Creatinine > 3.39 mg/dl, and a grade 3 or 4 hepatic encephalopathy should be considered as poor prognosis criteria in the patient. To avoid the need for liver transplantation associated with end-stage liver disease as the most available treatment nowadays, attention to the other new therapeutics approach such as: procalcitonin serum levels, vitamin K1, sustained c-Jun N-terminal kinase (JNK) activation, targeted metabolomics method based on the metabolic profiling of fatty acids, initial verification of keratin-18 and high-mobility group box-1 protein molecular, combination of adult stem cell transplantation with conservative medication recommended. Discussion:: The creation of N-acetyl-p-benzoquinone imine by cytochrome P450 and the generation of reactive oxygen species by paracetamol overdose could cause severe hepatic failure. The escalation in cellular and mitochondrial glutathione disulfide levels after paracetamol overdose suggests the participation of oxidative stress in the pathophysiology Conclusion:: With the advancements in technology and immunotherapies, the cost of liver transplantation continues to increase. Therefore, consideration should be given to further restrictions on paracetamol due to its availability as an over-the-counter drug.
2025-11-24 | Adipose-derived mesenchymal stem cell therapy modulates mitochondrial function to attenuate acetaminophen-induced liver injury by DDIT4/PGC-1α axis.
Acetaminophen (APAP) overdose is a leading cause of drug-induced liver injury, with limited treatment options. Mitochondrial dysfunction plays a central role in the pathogenesis of APAP-induced liver injury (AILI). Mitochondrial damage induces cell death, liver necrosis, and sever inflammation, leading to irreversible liver failure. Therefore, it is urgent to develop alternative treatments. Mesenchymal stem cell (MSC)-based cell therapy recently has received increasing attention for its role in regulating mitochondrial function. This study aimed to investigate the regulation of mitochondrial function and the therapeutic potential of MSCs in treating AILI and underlying mechanisms. AILI mouse model was established by injection of overdosed APAP. Mice were treated with adipose-derived MSCs (AMSCs) via the tail vein. The blood and liver tissues were collected for assessment of liver necrosis and mitochondrial function. To further verify the mechanism, hepatocyte-specific DNA damage-inducible transcript 4 (DDIT4) knockout (Ddit4△Hep) mice were generated. We showed that AMSCs treatment significantly reduced liver necrosis, oxidative stress, and mitochondrial dysfunction in AILI. DDIT4 expression in hepatocytes was identified as playing an important part in AMSCs treating AILI by using RNA-Seq and histological analysis. DDIT4 upregulation in liver samples of AILI patients characterized by snRNA-Seq and spatial transcriptomics analysis, which indicates DDIT4 plays a role in AILI. Ddit4△Hep mice exhibited exacerbated mitochondrial dysfunction and liver damage to APAP overdose, and these changes could not be reversed by AMSCs treatment. Mechanistically, AMSCs induced mitochondrial biogenesis and mitophagy through DDIT4-mediated PGC-1α upregulation, consequently leading to the restoration of mitochondrial mass and function in AILI. Finally, inhibition of PGC-1α abolished the protective effects of AMSCs against APAP-induced mitochondrial damage. In summary, this study indicates the potential role of stem cell therapy in modulating mitochondrial function and highlights the role of activation the DDIT4/PGC-1α pathway in protecting hepatic mitochondria to alleviate AILI. This study provides a new mechanistic perspective for stem cell therapy in the treatment of AILI and potential targets for clinical drug development.
2025-11-08 | Discovery of intrahepatic CD103+ cDC1/CD8+ TRM protective immune axis against acetaminophen-induced acute liver injury.
Understanding the intrahepatic protective immune systems against acetaminophen (APAP)-induced acute liver injury (ALI) is currently limited. Here we reveal that Gram-positive gut-microbiota-derived pathogen-associated molecular patterns promote the CCL2-dependent infiltration of hepatotoxic Ly6Chi monocytes into the APAP-damaged liver, thus inducing APAP-ALI. Conversely, Gram-negative bacterial pathogen-associated molecular patterns activate hepatic CD103⁺ cDC1s to produce IL-15, which in turn expands intrahepatic tissue-resident memory CD8⁺ T (TRM) cells and promotes protective immunity against APAP-derived liver injury. APAP-ALI was further exacerbated in Batf3-knockout and Rag1-knockout mice owing to an increased population of intrahepatic Ly6Chi monocytes in both knockout strains. The adoptive transfer of hepatic CD8+ T cells or hepatic CD103+ cDC1s from wild-type mice ameliorated APAP-ALI in both knockout mice. Notably, CD44+CD69+ TRM cells within hepatic CD8+ T cells, when activated by IL-15/IL-15Rα from hepatic CD103+ cDC1s of APAP mice, played a crucial role in inducing apoptosis of liver-infiltrating monocytes through direct cell-to-cell interactions and granzyme B secretion. Human results supported these animal findings. Our findings underscore the existence of an intrahepatic protective immune system, the hepatic CD103+ cDC1/CD8+ TRM axis, which regulates APAP-ALI by controlling pathogenic monocytes.
2025-10-14 | Microperfused Biomimetic Liver-on-a-Chip for High-Throughput Hepatotoxicity Screening.
Liver-on-a-chip has emerged as an effective tool for liver disease modeling and new drug development, while the current challenge lies in further mimicking liver physiological conditions. Here, inspired by the intricate architecture of hepatic lobules, a novel biomimetic vascularized liver-on-a-chip fabricated via high-precision 3D printing technology is presented. The chip features a capillary network with multiple micro-pores, ensuring a constant nutrient supply and efficient substance exchange within the cell environment through microfluidic perfusion. Under microperfusion conditions, human induced pluripotent stem cell-derived hepatocytes (hiPSC-Heps) exhibit superior physiological status and biological functionality compared to traditional 2D cultures. In addition, a comprehensive platform for assessing drug-induced liver injury (DILI) is established by integrating a concentration gradient chip, liver chips, and multicellular coculture technologies. This innovative platform effectively validates both acute and chronic hepatotoxic effects of acetaminophen (APAP). These findings demonstrate that the biomimetic vascularized liver-on-a-chip exhibits ideal physiological relevance and holds significant potential for applications in drug development and toxicity screening.
antibodies
2025-11-25 | An intrabody targeting PHD2 protects mice against acetaminophen-induced liver injury by attenuating ferroptosis.
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.
2025-08-17 | Platelets mediate neutrophil infiltration and exacerbate liver injury and sinusoidal endothelial cell damage after a severe acetaminophen overdose in mice.
Acetaminophen (APAP) overdose, the leading cause of acute liver failure in the US, is accompanied by hepatocyte necrosis and liver sinusoidal endothelial cell (LSEC) damage, with hepatic neutrophil infiltration and platelet deposition. However, the exact role played by platelets in the pathophysiology is not fully understood. To investigate this, we depleted platelets in C57Bl/6 J mice by injecting 2 mg/kg anti-CD41 antibody or IgG control 12 and 2 h before a moderate (300 mg/kg) or severe (600 mg/kg) APAP overdose. Platelet depletion did not affect liver injury as measured by plasma ALT levels and hepatic areas of necrosis 24 h after a moderate APAP overdose, but reduced hepatic neutrophil infiltration. After a severe overdose, platelet depletion caused a significant reduction in hepatic neutrophil infiltration, but contrary to the moderate overdose, it also significantly reduced liver injury. These findings were confirmed with a second platelet-depleting antibody (CD42b antibody). These results corroborate our previous findings, which showed that neutrophils exacerbate liver injury only after a severe APAP overdose but not a moderate one. Furthermore, severe APAP overdose induced LSEC injury 24 h after APAP, which was significantly reduced in platelet-depleted mice as measured by intrahepatic hemorrhage, increased hepatic von Willebrand factor deposition, and elevated plasma levels of hyaluronan. Thus, platelets contribute to endothelial cell injury and mediate neutrophil recruitment, which aggravates liver injury after a severe APAP overdose, underscoring the role of platelets in the pathophysiology.
2025-04-12 | Modular Nanotransporters Containing Keap1 Monobodies Are Capable of Reducing the Toxic Effect of Acetaminophen on the Liver of Mice.
Previously, we created a modular nanotransporter (MNT) containing a monobody to Keap1, an intracellular protein inhibitor of the Nrf2 transcription factor that controls cellular protection from oxidative stress and is capable of interacting with Keap1 in hepatocytes and protect these cells from the effects of hydrogen peroxide. Oxidative liver damage by acetaminophen was used as a model to study the antitoxic effect of this MNT. Intraperitoneal injection of acetaminophen to mice resulted in an increase in the level of alanine aminotransferase and aspartate aminotransferase in the blood, as well as in liver edema. A significant decrease in the level of these enzymes in the blood, along with a decrease in liver edema, was observed after preliminary intravenous administration of MNT 2 h before the acetaminophen injection. The results obtained can be used as a basis for developing drugs to treat diseases associated with oxidative stress.
2025-01-20 | Dynamic analysis of intrahepatic T cells reveals a unique group of restorative Cxcr3+ tissue-resident CD4 T cells in acute liver injury.
Acetaminophen (APAP) stands as one of the most prevalent triggers of drug-induced acute liver injury (ALI). The intricate modulation of immune system activation and inflammatory cascades by hepatic immune cells is paramount in managing liver injury and subsequent restoration. In this study, we employed an integrative approach that fused our proprietary flow cytometry analyses across various time points post-APAP injury with publicly available single-cell RNA sequencing (scRNA-seq) datasets, encompassing time-series data from liver tissue of mice subjected to APAP intoxication. This allowed us to delve into the dynamics of T cell profiles during APAP-induced ALI. Our comprehensive analyses unveiled the intricate temporal shifts in intrahepatic T cell populations across different phases following APAP-induced ALI. Notably, we observed a persistent augmentation of intrahepatic CD4+ T cells post-APAP injury. Amongst these, a distinct population of restorative Cxcr3+ tissue-resident CD4+ T cells emerged. Inhibition of CXCR3 using a neutralizing antibody exacerbated APAP-induced liver function impairment and hepatocyte death. Furthermore, we identified that the Cxcr3+ tissue-resident CD4+ T cells were tightly regulated by intrahepatic ''Lgals9-Cd45'' and 'CXCL13-Cxcr3' signaling pathways. These discoveries underscore the novel protective function of CXCR3, a vital biological macromolecule, in mitigating APAP-induced ALI, and may shed lights on new therapeutic strategies targeting this condition.
2024-08-20 | Both TREM2-dependent macrophages and Kupffer cells play a protective role in APAP-induced acute liver injury.
The inflammatory response is a significant factor in acetaminophen (APAP)-induced acute liver injury. And it can be mediated by macrophages of different origins. However, whether Kupffer cells and mononuclear-derived macrophages play an injury or protective role in APAP hepatotoxicity is still unclear. In this study, C57/BL6N mice were performed to establish the APAP acute liver injury model. Intervention experiments were also carried out using clodronate liposomes or TREM2 knockout. We found that APAP overdose triggered the activation of inflammatory factors and enhanced the expression of the RIPK1-MLKL pathway in mice's livers. Moreover, our study showed that inflammation-related protein expression was increased after clodronate liposome administration or TREM2 knockout. The RIPK1-MLKL-mediated necroptosis was also significantly activated after the elimination of Kupffer cells or the inhibition of mononuclear-derived macrophages. More importantly, clodronate liposomes treatment and TREM2 deficiency all worsen APAP-induced liver damage in mice. In conclusion, the results indicate that Kupffer cells and mononuclear macrophages play a protective role in APAP-induced liver injury by regulating necroptosis. Therefore, macrophages hold as a potential therapeutic target for APAP-induced liver damage.
proteins
2026-05-01 | DNAJB4/HLJ1 protects against acetaminophen-induced liver injury by attenuating ER stress via HSP70.
Drug-induced liver injury (DILI) remains a leading cause of acute liver failure; however, current clinical strategies lack reliable biomarkers, predictors of susceptibility, and effective therapeutic interventions. Among these etiologies, acetaminophen (APAP) overdose is the most common cause of DILI worldwide. Heat shock proteins (HSPs), particularly members of the HSP40 family, are central regulators of cellular stress responses, yet the specific role of DNAJB4/HLJ1 in APAP-induced hepatotoxicity remains poorly defined. To address this gap, we employed DNAJB4/HLJ1-deficient mice (Dnajb4-/-) to investigate the function of DNAJB4/HLJ1 in APAP-induced liver injury. Following APAP administration (> 400 mg/kg), Dnajb4-/- exhibited exacerbated hepatic necrosis, elevated liver enzymes, and enhanced c-jun/JNK activation compared with Dnajb4+/+ controls. Metabolic profiling revealed altered APAP metabolism, with reduced detoxification products and excessive oxidative metabolites, and pronounced glutathione (GSH) depletion. Transcriptomic analysis implicated DNAJB4/HLJ1 in metabolism, protein folding, and endoplasmic reticulum (ER) stress via interaction with HSP70. Consistently, ATF6, XBP1, and CHOP expression confirmed aggravated ER stress in Dnajb4-/- livers. AlphaFold-Multimer modeling and co-immunoprecipitation validated physical interaction between DNAJB4/HLJ1 and HSP70. Restoration of DNAJB4/HLJ1 expression attenuated ER stress, c-jun/JNK activation, and liver injury, while pharmacological inhibition of ER stress confirmed its mechanistic involvement. Collectively, these findings identify DNAJB4/HLJ1 as a previously unrecognized regulator of stress signaling in APAP-induced hepatotoxicity. By modulating ER stress within the integrated cellular stress network, DNAJB4/HLJ1 limits injury progression and promotes hepatocellular resilience, highlighting its potential as a novel therapeutic target for preventing or mitigating DILI.
2026-04-22 | Protective effect of latex proteins from Plumeria pudica against acetaminophen-induced acute liver injury.
The protein fraction from Plumeria pudica latex (LPPp) was evaluated against APAP-induced liver injury in mice. Key measurements included total leukocyte count, liver weight, AST, ALT, liver levels of MDA, GSH, SOD, MPO, NO3/NO2 concentration, cytokines TNF-α, IL-β, IL-6, IL-10, IFN- γ, and CXCL1. Histopathological and immunohistochemical analysis were conducted, and proteins in LPPp were identified by mass spectrometry. Molecular docking analyses were performed with LPPp proteins and APAP, NAPQI, TNF-α, and IFN-α. LPPp reduced liver weight, ALT, and AST levels, and prevented the decrease in total leukocyte count, indicating protection against liver injury. It also reduced MDA levels and preserved GSH and SOD levels, showing its antioxidant properties. LPPp significantly inhibited MPO and NO3/NO2 levels, reducing inflammation. LPPp decreased levels of TNF-α, IL-β, IL-6, IFN-γ, and CXCL1, while IL-10 levels remained elevated, suggesting an anti-inflammatory response. Histological analysis showed preserved liver tissue, and anti-Ly6C immunostaining indicated reduced neutrophil accumulation. Molecular docking revealed that latex cysteine proteases had a higher affinity for APAP and NAPQI, while a proteinase inhibitor showed affinity for CYP2EI, TNF-α, and IFN-α. These findings suggest that LPPp protects against APAP hepatotoxicity by interfering with APAP and NAPQI metabolism, reducing oxidative stress and inflammation, thus preventing hepatocyte damage.
2026-03-16 | PLGA nanoparticles encapsulating fibroblast growth factor 21 alleviate acetaminophen-induced acute liver injury.
The short half-life of fibroblast growth factor 21 (FGF21) in vivo limits its clinical application in acetaminophen (APAP)-induced acute liver injury (ALI). This study aimed to construct a novel PLGA-based nanoparticle delivery system for FGF21 and to investigate its protective effects and underlying mechanisms in APAP-induced ALI. PLGA-FGF21 nanoparticles were prepared using a double-emulsion solvent evaporation method and characterized by transmission electron microscopy and Zetasizer Nano for morphology, size, and stability. Their release profile and cytotoxicity were evaluated in vitro using AML12 cells, while hepatoprotective effects were assessed in an APAP-induced ALI mouse model. Histopathology, biochemical assays, immunofluorescence, and Western blot were employed to examine liver function, oxidative stress, and molecular mechanisms. The nanoparticles displayed uniform spherical morphology with an average size of 222.04 ± 1.08 nm and remained stable for 7 days in aqueous suspension. Encapsulation conferred sustained-release properties, and the particles showed no cytotoxicity while maintaining biological activity for at least 28 days. In vivo, PLGA-FGF21 significantly alleviated APAP-induced liver damage, reduced serum ALT and AST, and exhibited superior hepatoprotective efficacy compared with free FGF21. Furthermore, it suppressed inflammatory responses by decreasing TNF-α and MDA levels, while enhancing antioxidant defenses through increased SOD activity. Mechanistically, PLGA-FGF21 activated the PGC-1α/Nrf2/HO-1 signaling pathway, contributing to enhanced anti-inflammatory and antioxidant effects. PLGA-FGF21 nanoparticles provide potent protection against APAP-induced ALI by reducing inflammation and oxidative stress, which may be associated with activation of the PGC-1α/Nrf2 signaling pathway, offering a promising strategy for FGF21-based therapy in liver injury.
2026-03-01 | MOTS-c Protects Against Acetaminophen-induced Liver Injury through the MAPK Signaling Pathway.
Acetaminophen (APAP)-induced liver injury (AILI) is a leading cause of acute liver failure worldwide, but effective therapeutic strategies are still lacking. MOTS-c, a mitochondrial-derived peptide, has demonstrated hepatoprotective properties in models of nonalcoholic steatohepatitis (NASH) and hepatitis B virus (HBV) infection. This study aims to explore the role and underlying mechanisms of MOTS-c in AILI. An AILI model was established in male C57BL/6 mice via intraperitoneal (i.p.) injection of APAP (300 mg/kg). The therapeutic potential of MOTS-c and its mechanisms were assessed using behavioral tests, qPCR, western blotting, ELISA, immunohistochemistry, immunofluorescence, and TUNEL staining. MOTS-c levels in both plasma and liver tissues were significantly reduced in APAPinduced AILI mice compared with controls. Administration of MOTS-c via i.p. injection markedly attenuated APAP-induced increases in AST and ALT levels, histopathological liver damage, and other liver injury markers. MOTS-c treatment suppressed the release of pro-inflammatory factors (TNF-α, IL-1β, IL-6, and COX-2) and macrophage infiltration induced by APAP. Furthermore, MOTS-c treatment significantly restored GSH content, diminished reactive oxygen species (ROS) production, and oxidative stress. TUNEL staining confirmed that increased apoptosis in APAPtreated livers was significantly attenuated by MOTS-c, which are key contributors to hepatocyte death and liver injury. Mechanistic studies revealed that MOTS-c inhibited APAP-induced phosphorylation of MAPK pathway components, including ERK, JNK, and p38. The protective effects of MOTS-c on serum ALT and AST levels were abolished by co-treatment with inhibitors of ERK, JNK, and p38. This study reveals that the mitochondrial peptide MOTS-c can alleviate drug-induced liver injury by suppressing oxidative stress and inflammation via the MAPK pathway. This positions MOTS-c as a promising therapeutic candidate for treating APAP-induced liver injury. This study demonstrates that administering MOTS-c effectively protects against APAP-induced liver injury in mice. The protective mechanism involves suppressing the damaging MAPK signaling pathway (ERK, JNK, p38), which in turn reduces oxidative stress, inflammation, and cell death.
2026-02-19 | Annexin A1 enhances liver repair after acetaminophen-induced liver injury by regulating neutrophils function.
Acetaminophen (APAP)-induced acute liver injury (AILI) is a major cause of acute liver failure (ALF), for which effective and timely treatments remain limited. The mechanisms that drive irreversible progression to ALF are not fully understood. Annexin A1 (ANXA1), an anti-inflammatory factor, is implicated in several inflammatory diseases, but its role in APAP-induced liver injury is unclear. In this study, we establish a clinically relevant mouse model of APAP-induced AILI and show that ANXA1 expression increases in the liver, primarily originating from infiltrating neutrophils. ANXA1 deficiency exacerbated liver damage and hindered tissue repair, partly due to the formation of neutrophil extracellular traps (NETs) and altered neutrophil phenotypes. Inhibition of NETs by DNase I restored liver repair and promoted a shift of neutrophils toward an N2 anti-inflammatory phenotype in ANXA1-deficient mice. Furthermore, treatment with Ac2-26, an ANXA1-derived peptide, suppressed NET formation and modulated neutrophil phenotypes to alleviate liver injury. These findings suggest that ANXA1 facilitates liver repair after APAP-induced injury by reprogramming neutrophils via NETs inhibition, offering a promising therapeutic target to enhance recovery from drug-induced liver injury.
other
2025-08-27 | CpG-oligodeoxynucleotides challenged macrophages ameliorate acetaminophen induced liver injury by activating TLR9/IRG1/itaconate metabolic pathway.
Acetaminophen, or N-acetyl-para-aminophenol (APAP), causes severe liver damage and acute liver failure when overdosed. Oligodeoxynucleotides containing CpG motifs (CpG ODN) can regulate the function of macrophages, which play an important role in drug-induced liver injury. It is unclear whether CpG ODN-treated macrophages play an immune regulation role in APAP-induced liver injury. In the present study, we aim to explore the role of CpG ODN-activated macrophages in APAP-induced liver injury and the underlying mechanism in protecting against the cytotoxicity of APAP. In vivo, C57BL/6 mice were treated with APAP (300 mg/Kg) or/and CpG ODN (ODN 1826, 1.65 mg/Kg) by intraperitoneal injection, then survival rate, histopathological evaluation, and inflammatory factors were observed to ascertain the protective effect of CpG ODN. Then, CpG ODN-treated macrophages were reinfused into the animal model to determine the effector cells. In vitro, RNA sequencing and untargeted metabolomics detection were performed to illustrate the underlying mechanism. Last, Acod1 siRNA interference was used to clarify the role of IRG1 in resistance to APAP cytotoxicity by ROS and apoptosis indicator detections. We found that CpG ODN showed a protective effect against APAP cytotoxicity by stimulating macrophages rather than hepatic parenchymal cells. In particular, reinfusion of CpG ODN-treated macrophages to mice can alleviate APAP-induced liver injury. Transcriptome and metabolome analysis revealed that the expression of aconitate decarboxylase 1 (Acod1; also known as immune responsive gene 1, IRG1) and the metabolite itaconate generated by IRG1 catalysis increased after CpG ODN stimulation. In addition, we found that the mechanism of this protective effect is ascribed to the increased expression of Acod1 and the antioxidative function of itaconate by the activation of the TLR9/NF-κB signaling pathway. CpG ODN alleviated liver injury induced by APAP through the activation of the TLR9/NF-κB signaling pathway in macrophages, upregulating the expression of IRG1 protein, promoting the production of endogenous metabolite itaconate, and inhibiting macrophage apoptosis which was regulated by upregulating the expression of Nrf2 to inhibit ROS production. This study sheds new light on CpG ODN as a therapeutic strategy in resistance to APAP-induced liver injury.
2025-07-02 | miR-375 protects against acetaminophen-induced acute liver failure by orchestrating pharmacogene expression.
Acetaminophen (APAP) overdose is a leading cause of acute liver failure (ALF), primarily through the excessive production of N-acetyl-p-benzoquinone imine (NAPQI). N-acetylcysteine (NAC) is the Food and Drug Administration-approved treatment for APAP overdose, but there is a growing interest in microRNAs as potential therapeutic agents. We delivered miR-375 ectopically via a liver-tropic adeno-associated virus serotype 8 (AAV8) and demonstrated its potent protection in a murine model of APAP overdose-induced ALF. Slc16a2, Cyb5b, and Acsl5 were identified as critical targets acting synergistically to mitigate toxicity. Liver transcriptome revealed that miR-375 overexpression or silencing of the targets of miR-375 increased Gstm3 expression in mice. AAV8-mediated Gstm3 expression protects against APAP-ALF, and the protection was further enhanced by disrupting the expression of Cyp2e1. Additionally, CYP2E1 and GSS, which contribute to APAP detoxification, were down- and upregulated by miR-375, respectively. These findings suggest that miR-375 prevents APAP-ALF by orchestrating the expression of pharmacogenes and enhancing glutathione synthesis. We conclude that miR-375 and its targets are promising therapeutic targets for APAP-ALF.
2025-02-28 | Liver-specific Bcl3 Knockout Alleviates Acetaminophen-induced Liver Injury by Activating Nrf2 Pathway in Male Mice.
Acetaminophen (APAP) overdose is the leading cause of acute liver failure, with oxidative stress being a critical factor in this process. Glutathione (GSH) plays a vital defensive role. Activation of nuclear factor erythroid 2 like 2 (Nrf2) pathway mitigates APAP-induced liver damage by promoting GSH biosynthesis and enhancing drug detoxification. Although the role of B cell leukemia/lymphoma 3 (Bcl3) in regulating inflammatory responses, cellular oncogenesis, and immune balance is well-documented, its function in APAP-induced liver injury remains unclear. We employed liver-specific Bcl3 knockout (Bcl3hep-/-) mice and adeno-associated virus (AAV)-8-mediated Bcl3 overexpression (AAV-Bcl3) mice to model APAP-induced liver injury. Liver damage was assessed through hematoxylin and eosin staining and serum alanine aminotransferase and aspartate aminotransferase measurements. The interaction between Bcl3 and Nrf2 was examined using immunofluorescence and co-immunoprecipitation assays. Our study reveals a significant upregulation of Bcl3 expression in the livers of male mice following APAP administration, suggesting Bcl3's potential involvement in this pathological process. In Bcl3hep-/- mice, a reduced severity of liver damage was observed at both 6 and 24 hours post-APAP treatment compared with controls. Notably, Bcl3-deficient mice exhibited accelerated GSH replenishment due to the rapid induction of Gclc and Gclm genes following 6 hours of APAP exposure. Through immunofluorescence and co-immunoprecipitation analyses, we identified an interaction between Bcl3 and Nrf2. The loss of Bcl3 enhanced Nrf2 translocation upon APAP challenge, leading to the upregulation of antioxidant gene expression. These findings suggest that Bcl3 knockout alleviates oxidative stress resulting from APAP overdose. We uncovered a previously uncharacterized role of Bcl3 in APAP-induced liver injury, emphasizing the role of the Bcl3-Nrf2 axis in oxidative stress-related liver damage.
2024-07-28 | Adenosine kinase protects against acetaminophen-induced acute liver injury by activating autophagy in hepatocytes.
Acute liver injury (ALI) is a common life-threatening condition with a high mortality rate due to liver disease-related death. However, current therapeutic interventions for ALI remain ineffective, and the development of effective novel therapies is urgently needed. Liver samples from patients with drug-induced ALI were collected to detect adenosine kinase (ADK) expression. Male C57BL/6 J mice, hepatocyte-specific ADK knockout (ADKHKO) mice, and their controls (ADKf/f) were exposed to acetaminophen (APAP) and other treatments to investigate the mechanisms of APAP-related ALI. ADK expression was significantly decreased in APAP-injured livers. Hepatocyte-specific ADK deficiency exacerbated APAP-induced ALI, while a gain-of-function approach delivering AAV-ADK, markedly alleviated APAP-induced ALI, as indicated by changes in alanine aminotransferases (ALT) levels, aspartate aminotransferase (AST) levels, neutrophil infiltration and hepatocyte death. This study showed that ADK played a critical role in ALI by activating autophagy through two signaling pathways, the adenosine monophosphate-activated protein kinase (AMPK)-mTOR pathway and the adenosine receptor A1 (ADORA1)-Akt-mTOR pathway. Furthermore, we found that metformin upregulated ADK expression in hepatocytes and protected against APAP-induced ALI. These results demonstrate that ADK is critical in protecting against APAP-induced ALI and that developing therapeutics targeting ADK-adenosine-ADORA1 is a new approach for ALI treatment. Metformin is a potential candidate for preventing ALI by upregulating ADK.
2024-03-26 | Deletion of hepatocyte cysteine dioxygenase type 1, a bile acid repressed gene, enhances glutathione synthesis and ameliorates acetaminophen hepatotoxicity.
Liver is a major organ that metabolizes sulfur amino acids cysteine, which is the substrate for the synthesis of many essential cellular molecules including GSH, taurine, and coenzyme A. Bile acid-activated farnesoid x receptor (FXR) inhibits cysteine dioxygenase type 1 (CDO1), which mediates hepatic cysteine catabolism and taurine synthesis. To define the impact of bile acid inhibition of CDO1 on hepatic sulfur amino acid metabolism and antioxidant capacity, we developed hepatocyte-specific CDO1 knockout mice (Hep-CDO1 KO) and hepatocyte specific CDO1 transgenic mice (Hep-CDO1 Tg). Liver metabolomics revealed that genetic deletion of hepatic CDO1 reduced de novo taurine synthesis but had no impact on hepatic taurine abundance or bile acid conjugation. Consistent with reduced cysteine catabolism, Hep-CDO1 KO mice showed increased hepatic cysteine abundance but unaltered methionine cycle intermediates and coenzyme A synthesis. Upon acetaminophen overdose, Hep-CDO1 KO mice showed increased GSH synthesis capacity and alleviated liver injury. In contrast, hepatic CDO1 overexpression in Hep-CDO1 Tg mice stimulated hepatic cysteine to taurine conversion, resulting in reduced hepatic cysteine abundance. However, Hep-CDO1 Tg mice and WT showed similar susceptibility to acetaminophen-induced liver injury. Hep-CDO1 Tg mice showed similar hepatic taurine and coenzyme A compared to WT mice. In summary, these findings suggest that bile acid and FXR signaling inhibition of CDO1-mediated hepatic cysteine catabolism preferentially modulates hepatic GSH synthesis capacity and antioxidant defense, but has minimal effect on hepatic taurine and coenzyme A abundance. Repression of hepatic CDO1 may contribute to the hepatoprotective effects of FXR activation under certain pathologic conditions.
small molecules
2026-08-17 | Subchronic Alcohol Exposure Exacerbates Acetaminophen-Induced Acute Liver Injury via Macrophage-Mediated Inflammation and Mitochondrial Dysfunction.
Alcohol use is a major coexposure in patients taking acetaminophen (APAP), yet how subchronic alcohol exposure reshapes hepatic vulnerability to APAP remains incompletely defined. In this study, we examined the effects of chronic alcohol intake on APAP-induced liver injury in mice. Our results indicated that subchronic alcohol exposure significantly increased the mortality rate in APAP-overdosed mice. Alcohol pretreatment also exacerbated APAP-induced hepatic damage, as evidenced by elevated serum ALT and AST levels, enhanced inflammatory responses, and aggravated oxidative stress. Quantitative polymerase chain reaction (qPCR) analysis revealed that alcohol consumption suppressed mitochondrial biogenesis (MB)-related genes, including Pgc-1α and transcription factor A, mitochondrial (TFAM). Notably, macrophage depletion via clodronate liposomes not only attenuated hepatocyte injury and inflammation induction but also restored the expression of MB/function-related genes, including Pgc-1α, TFAM, and Opa1. These findings suggest macrophage-involved inflammatory amplification may aggravate APAP-induced liver injury, at least in part, by impairing mitochondrial adaptive repair responses.
2026-08-11 | Clinical Characteristics and Pattern of Paracetamol-Induced Liver Injury at Hospital Shah Alam: A Three-Year Retrospective Case Series
Background: Paracetamol poisoning is the most common type of therapeutic drug poisoning seen globally. The dose of ingested paracetamol and the time of presentation to a healthcare facility are important factors in assessing the severity of poisoning. In addition, factors affecting hepatic glutathione levels further influence the risk of toxicity. Objective: This three years case series aims to analyse the characteristics of liver injury observed in patients who presented to the hospital’s ED for Shah Alam following an intentional paracetamol overdose. Additionally, it seeks to determine risk factors specific to this local population. Methodology: Patients with a history of intentional paracetamol overdose and biochemical evidence of liver injury who presented to the ED of Hospital Shah Alam between 1st January 2022 and 31st December 2024 were included in this study. Patients aged less than 18 years were excluded from this study. Results: A total of 13 patients were included in this case series. The pattern of liver injury followed an established clinical course; however, no patients developed liver failure. High doses of paracetamol ingestion and late presentation after overdose were the expected risk factors. A further risk factor identified in this case series was low body weight. Six patients in this case series weighed less than 45 kg. Underweight patients may have underlying malnutrition. Paracetamol overdose hepatotoxicity risk is linked to depleted hepatic glutathione, which is known to be due to malnutrition. Conclusion: High doses of paracetamol ingestion with late presentation to a healthcare facility are known risk factors for paracetamol-induced liver injury. This case series also revealed that underweight patients are at risk, possibly because of malnutrition. These findings suggest that clinicians may consider a lower treatment threshold, such as the 100-line threshold, in underweight or potentially malnourished patients, although larger studies are needed.
2026-08-11 | Defining risk: the case for conceptual clarity in acetaminophen poisoning
Despite the widespread availability of acetylcysteine, acetaminophen (paracetamol) poisoning remains the most common cause of acute liver failure and acute liver transplantation in the United State...
2026-08-07 | Acetaminophen Poisoning: From Molecular Mechanisms to Multidisciplinary Management - A Contemporary Clinical Overview
Acetaminophen (N-acetyl-para-aminophenol, paracetamol) remains the leading cause of acute liver failure in the United States and accounts for a substantial proportion of pharmaceutical overdoses globally. This comprehensive review examines the epidemiology, pathophysiology, clinical presentation, and evidence-based management of acetaminophen toxicity, with particular emphasis on the critical role of timely intervention. The drug's widespread availability and pervasive public perception of safety contribute to its prominence in toxicity statistics, with over 60 million Americans using acetaminophen weekly, often unknowingly through combination products. The pathophysiological cascade is initiated by the saturation of hepatic sulfation and glucuronidation pathways, leading to the formation of the highly reactive metabolite N-acetyl-p-benzoquinoneimine (NAPQI) through cytochrome P450-mediated oxidation, predominantly via CYP2E1. When hepatic glutathione stores are depleted, NAPQI binds covalently to mitochondrial proteins, triggering oxidative stress, lipid peroxidation, and ultimately centrilobular necrosis. The clinical course classically progresses through four distinct stages, with the Rumack-Matthew nomogram serving as the cornerstone for risk stratification following acute ingestion. N-acetylcysteine, the specific antidote, is most effective when administered within 8 hours of ingestion and acts through glutathione replenishment, direct conjugation with NAPQI, and anti-inflammatory mechanisms. Emerging evidence suggests a potential role for fomepizole as adjunctive therapy in high-risk patients, though this remains investigational. Prognosis is excellent with early treatment, but delays significantly increase morbidity and mortality, with approximately 1-3% of severe cases requiring liver transplantation. A multidisciplinary approach encompassing emergency medicine, toxicology, hepatology, and critical care is essential for optimal patient outcomes. This review underscores the imperative for enhanced patient education, improved labeling practices, and continued research into novel therapeutic strategies to further reduce the burden of acetaminophen-induced hepatotoxicity.
2026-07-31 | Boosting Thioredoxin-1 Protects Against Acetaminophen-Induced Cardiotoxicity
Background: Although overdose-induced acute hepatic failure following acetaminophen (APAP) administration is well documented, clinical studies have also reported myocardial infarction, cardiac dysfunction, arrhythmias, pericarditis, and cardiac myocyte necrosis in APAP poisoning. As APAP overdose is associated with excessive reactive oxygen species (ROS) production and metabolism-mediated glutathione depletion, the cytosolic thioredoxin system (Trx-1/TrxR1/NADPH) may offer an alternative mechanism for cardioprotection. Although transgenic mice overexpressing thioredoxin-1 (Trx-1) are protected against doxorubicin/Adriamycin-induced cardiotoxicity, whether pharmacological enhancement of the cytosolic Trx system can mitigate APAP-induced cardiotoxicity and improve the therapeutic efficacy of chlorpromazine (CPZ) remains unclear. Methods: Cardiotoxicity was induced in wild-type C57BL/6 mice using APAP (400 mg/kg, i.p.). Expression of the Trx system was upregulated via chronic daily administration of low-dose epinephrine (EP, 0.2 mg/kg, s.c. for 14 days), and mice were subsequently treated with a subtoxic dose of CPZ (6 mg/kg, i.p.). Results: EP significantly upregulated the expression of Trx-1, thioredoxin-like protein 1 (Trx-L1), and thioredoxin reductase 1 (TrxR1) in cardiac tissue. Furthermore, EP attenuated APAP-induced acute cardiac injury, and its combination with CPZ additively amplified these cardioprotective effects. These findings were supported by the normalization of serum cardiac biomarkers (aspartate aminotransferase [AST], lactate dehydrogenase [LDH], and cardiac troponin I [cTnI]), reduction of oxidative stress (restoration of reduced glutathione [GSH], superoxide dismutase [SOD], and nuclear factor erythroid 2-related factor 2 [Nrf2]) and inflammatory markers (tumor necrosis factor-alpha [TNF-α]) in cardiac tissue, and preservation of normal myocardial histological architecture. Mechanistically, activation of the Trx system was accompanied by upregulation of β-arrestin-1 and cAMP response element-binding protein 1 (CREB1), indicating their involvement in Trx-mediated cardioprotection. Conclusion: Pharmacological enhancement of the cytosolic thioredoxin system protects against APAP-induced acute cardiac injury, and combining EP with CPZ exerts an additive cardioprotective effect.
cell therapies
2026-07-21 | Adoptive transfer of VSIG4+ macrophages interrupts the CCL2-CCR2 inflammatory amplification loop to mitigate acetaminophen-induced acute liver injury in murine and human organoid models.
Acute liver injury (ALI) can rapidly progress to life-threatening acute liver failure. Liver transplantation remains the only definitive treatment, despite critical donor shortages. Macrophage-based therapies have shown promise in ALI but face challenges related to subset imprecision and limited humanized validation. VSIG4 is generally considered a marker of Kupffer cells; however, it is also expressed in monocyte-derived macrophages (MoMFs), and its role remains unclear during ALI. By integrating single-cell RNA sequencing (scRNA-seq) data from human and mouse liver tissues with clinical ALI tissue samples, we elucidated the dynamic changes in VSIG4+ macrophages (VSIG4+ Mφ) within the liver. We developed a reversible immunomagnetic nanoparticle system for the non-destructive isolation of viable VSIG4+ Mφ. The therapeutic efficacy and potential mechanisms of VSIG4+ Mφ were evaluated through tissue and molecular-level analyses in an acetaminophen (APAP)-induced ALI mouse model, as well as in a newly established vascularized human liver organoid ALI and monocyte chemotaxis model. scRNA-seq revealed a previously underrecognized subpopulation of VSIG4+ MoMFs, which increases following ALI, while the number of resident VSIG4+ Kupffer cells decreases significantly. Clinical ALI tissue samples also confirmed the presence of CCR2+VSIG4+ cells in the livers of patients with ALI. In the APAP-induced ALI animal model, adoptive transfer of isolated VSIG4+ MoMFs dramatically decreased serum alanine transaminase, hepatic necrosis, and apoptosis while increasing anti-inflammatory cytokines. In contrast, adoptive transfer of unselected BMDM failed to improve liver injury and instead exacerbated certain pro-inflammatory responses, including elevated TNF-α and reduced CD206 expression. Mechanistically, VSIG4+ MoMFs prevented inflammatory amplification by suppressing NF-κB-dependent CCL2 transcription, thereby disrupting the CCL2-CCR2 chemotactic axis and reducing pro-inflammatory CCR2+ monocyte and macrophage recruitment. We further developed vascularized human liver organoids and APAP-induced hepatocyte injury and monocyte chemotaxis, finding that the chemotaxis-interrupting mechanism was fully recapitulated in the human liver organoid ALI model. This study identifies VSIG4+ MoMFs as a therapeutically viable subset for ALI, with clear superiority over unselected BMDM. By blocking the CCL2-CCR2 inflammatory amplification loop, these cells attenuate liver injury in both mouse and humanized models. These consistent findings provide robust preclinical evidence to support the advancement of VSIG4+ Mφ-based immunotherapies into clinical practice.
2026-03-18 | Paracetamol Poisoning and Liver Transplantation: An Updated Review
Introduction:: Regarding clinical recovery and survival of patients with severe paracetamol- induced acute liver failure and hepatic encephalopathy, liver transplantation is recommended as a good approach to management. This review summarizes the findings of previous studies associated with paracetamol-induced hepatotoxicity and liver transplantation. Methods:: All relevant articles (n=113) were searched systematically between the relevant record initiation and 14 October 2025 (the date of our final search), across PubMed and Web of Science. The search terms included the main concepts and their synonyms, including the problem: “paracetamol- over-dose”, “paracetamol poisoning”, the intervention “assessment of hepatotoxicity”, and the outcome “liver transplantation”, combined with the AND, OR operators. Synonyms for this concept were extracted from thesauri such as Medical Subject Headings (MeSH). Results:: An arterial pH lower than 7.3 or all three of the following: 1) International Normalized ratio of Prothrombin Time > 6.5, 2) Creatinine > 3.39 mg/dl, and a grade 3 or 4 hepatic encephalopathy should be considered as poor prognosis criteria in the patient. To avoid the need for liver transplantation associated with end-stage liver disease as the most available treatment nowadays, attention to the other new therapeutics approach such as: procalcitonin serum levels, vitamin K1, sustained c-Jun N-terminal kinase (JNK) activation, targeted metabolomics method based on the metabolic profiling of fatty acids, initial verification of keratin-18 and high-mobility group box-1 protein molecular, combination of adult stem cell transplantation with conservative medication recommended. Discussion:: The creation of N-acetyl-p-benzoquinone imine by cytochrome P450 and the generation of reactive oxygen species by paracetamol overdose could cause severe hepatic failure. The escalation in cellular and mitochondrial glutathione disulfide levels after paracetamol overdose suggests the participation of oxidative stress in the pathophysiology Conclusion:: With the advancements in technology and immunotherapies, the cost of liver transplantation continues to increase. Therefore, consideration should be given to further restrictions on paracetamol due to its availability as an over-the-counter drug.
2025-11-24 | Adipose-derived mesenchymal stem cell therapy modulates mitochondrial function to attenuate acetaminophen-induced liver injury by DDIT4/PGC-1α axis.
Acetaminophen (APAP) overdose is a leading cause of drug-induced liver injury, with limited treatment options. Mitochondrial dysfunction plays a central role in the pathogenesis of APAP-induced liver injury (AILI). Mitochondrial damage induces cell death, liver necrosis, and sever inflammation, leading to irreversible liver failure. Therefore, it is urgent to develop alternative treatments. Mesenchymal stem cell (MSC)-based cell therapy recently has received increasing attention for its role in regulating mitochondrial function. This study aimed to investigate the regulation of mitochondrial function and the therapeutic potential of MSCs in treating AILI and underlying mechanisms. AILI mouse model was established by injection of overdosed APAP. Mice were treated with adipose-derived MSCs (AMSCs) via the tail vein. The blood and liver tissues were collected for assessment of liver necrosis and mitochondrial function. To further verify the mechanism, hepatocyte-specific DNA damage-inducible transcript 4 (DDIT4) knockout (Ddit4△Hep) mice were generated. We showed that AMSCs treatment significantly reduced liver necrosis, oxidative stress, and mitochondrial dysfunction in AILI. DDIT4 expression in hepatocytes was identified as playing an important part in AMSCs treating AILI by using RNA-Seq and histological analysis. DDIT4 upregulation in liver samples of AILI patients characterized by snRNA-Seq and spatial transcriptomics analysis, which indicates DDIT4 plays a role in AILI. Ddit4△Hep mice exhibited exacerbated mitochondrial dysfunction and liver damage to APAP overdose, and these changes could not be reversed by AMSCs treatment. Mechanistically, AMSCs induced mitochondrial biogenesis and mitophagy through DDIT4-mediated PGC-1α upregulation, consequently leading to the restoration of mitochondrial mass and function in AILI. Finally, inhibition of PGC-1α abolished the protective effects of AMSCs against APAP-induced mitochondrial damage. In summary, this study indicates the potential role of stem cell therapy in modulating mitochondrial function and highlights the role of activation the DDIT4/PGC-1α pathway in protecting hepatic mitochondria to alleviate AILI. This study provides a new mechanistic perspective for stem cell therapy in the treatment of AILI and potential targets for clinical drug development.
2025-11-08 | Discovery of intrahepatic CD103+ cDC1/CD8+ TRM protective immune axis against acetaminophen-induced acute liver injury.
Understanding the intrahepatic protective immune systems against acetaminophen (APAP)-induced acute liver injury (ALI) is currently limited. Here we reveal that Gram-positive gut-microbiota-derived pathogen-associated molecular patterns promote the CCL2-dependent infiltration of hepatotoxic Ly6Chi monocytes into the APAP-damaged liver, thus inducing APAP-ALI. Conversely, Gram-negative bacterial pathogen-associated molecular patterns activate hepatic CD103⁺ cDC1s to produce IL-15, which in turn expands intrahepatic tissue-resident memory CD8⁺ T (TRM) cells and promotes protective immunity against APAP-derived liver injury. APAP-ALI was further exacerbated in Batf3-knockout and Rag1-knockout mice owing to an increased population of intrahepatic Ly6Chi monocytes in both knockout strains. The adoptive transfer of hepatic CD8+ T cells or hepatic CD103+ cDC1s from wild-type mice ameliorated APAP-ALI in both knockout mice. Notably, CD44+CD69+ TRM cells within hepatic CD8+ T cells, when activated by IL-15/IL-15Rα from hepatic CD103+ cDC1s of APAP mice, played a crucial role in inducing apoptosis of liver-infiltrating monocytes through direct cell-to-cell interactions and granzyme B secretion. Human results supported these animal findings. Our findings underscore the existence of an intrahepatic protective immune system, the hepatic CD103+ cDC1/CD8+ TRM axis, which regulates APAP-ALI by controlling pathogenic monocytes.
2025-10-14 | Microperfused Biomimetic Liver-on-a-Chip for High-Throughput Hepatotoxicity Screening.
Liver-on-a-chip has emerged as an effective tool for liver disease modeling and new drug development, while the current challenge lies in further mimicking liver physiological conditions. Here, inspired by the intricate architecture of hepatic lobules, a novel biomimetic vascularized liver-on-a-chip fabricated via high-precision 3D printing technology is presented. The chip features a capillary network with multiple micro-pores, ensuring a constant nutrient supply and efficient substance exchange within the cell environment through microfluidic perfusion. Under microperfusion conditions, human induced pluripotent stem cell-derived hepatocytes (hiPSC-Heps) exhibit superior physiological status and biological functionality compared to traditional 2D cultures. In addition, a comprehensive platform for assessing drug-induced liver injury (DILI) is established by integrating a concentration gradient chip, liver chips, and multicellular coculture technologies. This innovative platform effectively validates both acute and chronic hepatotoxic effects of acetaminophen (APAP). These findings demonstrate that the biomimetic vascularized liver-on-a-chip exhibits ideal physiological relevance and holds significant potential for applications in drug development and toxicity screening.
antibodies
2025-11-25 | An intrabody targeting PHD2 protects mice against acetaminophen-induced liver injury by attenuating ferroptosis.
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.
2025-08-17 | Platelets mediate neutrophil infiltration and exacerbate liver injury and sinusoidal endothelial cell damage after a severe acetaminophen overdose in mice.
Acetaminophen (APAP) overdose, the leading cause of acute liver failure in the US, is accompanied by hepatocyte necrosis and liver sinusoidal endothelial cell (LSEC) damage, with hepatic neutrophil infiltration and platelet deposition. However, the exact role played by platelets in the pathophysiology is not fully understood. To investigate this, we depleted platelets in C57Bl/6 J mice by injecting 2 mg/kg anti-CD41 antibody or IgG control 12 and 2 h before a moderate (300 mg/kg) or severe (600 mg/kg) APAP overdose. Platelet depletion did not affect liver injury as measured by plasma ALT levels and hepatic areas of necrosis 24 h after a moderate APAP overdose, but reduced hepatic neutrophil infiltration. After a severe overdose, platelet depletion caused a significant reduction in hepatic neutrophil infiltration, but contrary to the moderate overdose, it also significantly reduced liver injury. These findings were confirmed with a second platelet-depleting antibody (CD42b antibody). These results corroborate our previous findings, which showed that neutrophils exacerbate liver injury only after a severe APAP overdose but not a moderate one. Furthermore, severe APAP overdose induced LSEC injury 24 h after APAP, which was significantly reduced in platelet-depleted mice as measured by intrahepatic hemorrhage, increased hepatic von Willebrand factor deposition, and elevated plasma levels of hyaluronan. Thus, platelets contribute to endothelial cell injury and mediate neutrophil recruitment, which aggravates liver injury after a severe APAP overdose, underscoring the role of platelets in the pathophysiology.
2025-04-12 | Modular Nanotransporters Containing Keap1 Monobodies Are Capable of Reducing the Toxic Effect of Acetaminophen on the Liver of Mice.
Previously, we created a modular nanotransporter (MNT) containing a monobody to Keap1, an intracellular protein inhibitor of the Nrf2 transcription factor that controls cellular protection from oxidative stress and is capable of interacting with Keap1 in hepatocytes and protect these cells from the effects of hydrogen peroxide. Oxidative liver damage by acetaminophen was used as a model to study the antitoxic effect of this MNT. Intraperitoneal injection of acetaminophen to mice resulted in an increase in the level of alanine aminotransferase and aspartate aminotransferase in the blood, as well as in liver edema. A significant decrease in the level of these enzymes in the blood, along with a decrease in liver edema, was observed after preliminary intravenous administration of MNT 2 h before the acetaminophen injection. The results obtained can be used as a basis for developing drugs to treat diseases associated with oxidative stress.
2025-01-20 | Dynamic analysis of intrahepatic T cells reveals a unique group of restorative Cxcr3+ tissue-resident CD4 T cells in acute liver injury.
Acetaminophen (APAP) stands as one of the most prevalent triggers of drug-induced acute liver injury (ALI). The intricate modulation of immune system activation and inflammatory cascades by hepatic immune cells is paramount in managing liver injury and subsequent restoration. In this study, we employed an integrative approach that fused our proprietary flow cytometry analyses across various time points post-APAP injury with publicly available single-cell RNA sequencing (scRNA-seq) datasets, encompassing time-series data from liver tissue of mice subjected to APAP intoxication. This allowed us to delve into the dynamics of T cell profiles during APAP-induced ALI. Our comprehensive analyses unveiled the intricate temporal shifts in intrahepatic T cell populations across different phases following APAP-induced ALI. Notably, we observed a persistent augmentation of intrahepatic CD4+ T cells post-APAP injury. Amongst these, a distinct population of restorative Cxcr3+ tissue-resident CD4+ T cells emerged. Inhibition of CXCR3 using a neutralizing antibody exacerbated APAP-induced liver function impairment and hepatocyte death. Furthermore, we identified that the Cxcr3+ tissue-resident CD4+ T cells were tightly regulated by intrahepatic ''Lgals9-Cd45'' and 'CXCL13-Cxcr3' signaling pathways. These discoveries underscore the novel protective function of CXCR3, a vital biological macromolecule, in mitigating APAP-induced ALI, and may shed lights on new therapeutic strategies targeting this condition.
2024-08-20 | Both TREM2-dependent macrophages and Kupffer cells play a protective role in APAP-induced acute liver injury.
The inflammatory response is a significant factor in acetaminophen (APAP)-induced acute liver injury. And it can be mediated by macrophages of different origins. However, whether Kupffer cells and mononuclear-derived macrophages play an injury or protective role in APAP hepatotoxicity is still unclear. In this study, C57/BL6N mice were performed to establish the APAP acute liver injury model. Intervention experiments were also carried out using clodronate liposomes or TREM2 knockout. We found that APAP overdose triggered the activation of inflammatory factors and enhanced the expression of the RIPK1-MLKL pathway in mice's livers. Moreover, our study showed that inflammation-related protein expression was increased after clodronate liposome administration or TREM2 knockout. The RIPK1-MLKL-mediated necroptosis was also significantly activated after the elimination of Kupffer cells or the inhibition of mononuclear-derived macrophages. More importantly, clodronate liposomes treatment and TREM2 deficiency all worsen APAP-induced liver damage in mice. In conclusion, the results indicate that Kupffer cells and mononuclear macrophages play a protective role in APAP-induced liver injury by regulating necroptosis. Therefore, macrophages hold as a potential therapeutic target for APAP-induced liver damage.
proteins
2026-05-01 | DNAJB4/HLJ1 protects against acetaminophen-induced liver injury by attenuating ER stress via HSP70.
Drug-induced liver injury (DILI) remains a leading cause of acute liver failure; however, current clinical strategies lack reliable biomarkers, predictors of susceptibility, and effective therapeutic interventions. Among these etiologies, acetaminophen (APAP) overdose is the most common cause of DILI worldwide. Heat shock proteins (HSPs), particularly members of the HSP40 family, are central regulators of cellular stress responses, yet the specific role of DNAJB4/HLJ1 in APAP-induced hepatotoxicity remains poorly defined. To address this gap, we employed DNAJB4/HLJ1-deficient mice (Dnajb4-/-) to investigate the function of DNAJB4/HLJ1 in APAP-induced liver injury. Following APAP administration (> 400 mg/kg), Dnajb4-/- exhibited exacerbated hepatic necrosis, elevated liver enzymes, and enhanced c-jun/JNK activation compared with Dnajb4+/+ controls. Metabolic profiling revealed altered APAP metabolism, with reduced detoxification products and excessive oxidative metabolites, and pronounced glutathione (GSH) depletion. Transcriptomic analysis implicated DNAJB4/HLJ1 in metabolism, protein folding, and endoplasmic reticulum (ER) stress via interaction with HSP70. Consistently, ATF6, XBP1, and CHOP expression confirmed aggravated ER stress in Dnajb4-/- livers. AlphaFold-Multimer modeling and co-immunoprecipitation validated physical interaction between DNAJB4/HLJ1 and HSP70. Restoration of DNAJB4/HLJ1 expression attenuated ER stress, c-jun/JNK activation, and liver injury, while pharmacological inhibition of ER stress confirmed its mechanistic involvement. Collectively, these findings identify DNAJB4/HLJ1 as a previously unrecognized regulator of stress signaling in APAP-induced hepatotoxicity. By modulating ER stress within the integrated cellular stress network, DNAJB4/HLJ1 limits injury progression and promotes hepatocellular resilience, highlighting its potential as a novel therapeutic target for preventing or mitigating DILI.
2026-04-22 | Protective effect of latex proteins from Plumeria pudica against acetaminophen-induced acute liver injury.
The protein fraction from Plumeria pudica latex (LPPp) was evaluated against APAP-induced liver injury in mice. Key measurements included total leukocyte count, liver weight, AST, ALT, liver levels of MDA, GSH, SOD, MPO, NO3/NO2 concentration, cytokines TNF-α, IL-β, IL-6, IL-10, IFN- γ, and CXCL1. Histopathological and immunohistochemical analysis were conducted, and proteins in LPPp were identified by mass spectrometry. Molecular docking analyses were performed with LPPp proteins and APAP, NAPQI, TNF-α, and IFN-α. LPPp reduced liver weight, ALT, and AST levels, and prevented the decrease in total leukocyte count, indicating protection against liver injury. It also reduced MDA levels and preserved GSH and SOD levels, showing its antioxidant properties. LPPp significantly inhibited MPO and NO3/NO2 levels, reducing inflammation. LPPp decreased levels of TNF-α, IL-β, IL-6, IFN-γ, and CXCL1, while IL-10 levels remained elevated, suggesting an anti-inflammatory response. Histological analysis showed preserved liver tissue, and anti-Ly6C immunostaining indicated reduced neutrophil accumulation. Molecular docking revealed that latex cysteine proteases had a higher affinity for APAP and NAPQI, while a proteinase inhibitor showed affinity for CYP2EI, TNF-α, and IFN-α. These findings suggest that LPPp protects against APAP hepatotoxicity by interfering with APAP and NAPQI metabolism, reducing oxidative stress and inflammation, thus preventing hepatocyte damage.
2026-03-16 | PLGA nanoparticles encapsulating fibroblast growth factor 21 alleviate acetaminophen-induced acute liver injury.
The short half-life of fibroblast growth factor 21 (FGF21) in vivo limits its clinical application in acetaminophen (APAP)-induced acute liver injury (ALI). This study aimed to construct a novel PLGA-based nanoparticle delivery system for FGF21 and to investigate its protective effects and underlying mechanisms in APAP-induced ALI. PLGA-FGF21 nanoparticles were prepared using a double-emulsion solvent evaporation method and characterized by transmission electron microscopy and Zetasizer Nano for morphology, size, and stability. Their release profile and cytotoxicity were evaluated in vitro using AML12 cells, while hepatoprotective effects were assessed in an APAP-induced ALI mouse model. Histopathology, biochemical assays, immunofluorescence, and Western blot were employed to examine liver function, oxidative stress, and molecular mechanisms. The nanoparticles displayed uniform spherical morphology with an average size of 222.04 ± 1.08 nm and remained stable for 7 days in aqueous suspension. Encapsulation conferred sustained-release properties, and the particles showed no cytotoxicity while maintaining biological activity for at least 28 days. In vivo, PLGA-FGF21 significantly alleviated APAP-induced liver damage, reduced serum ALT and AST, and exhibited superior hepatoprotective efficacy compared with free FGF21. Furthermore, it suppressed inflammatory responses by decreasing TNF-α and MDA levels, while enhancing antioxidant defenses through increased SOD activity. Mechanistically, PLGA-FGF21 activated the PGC-1α/Nrf2/HO-1 signaling pathway, contributing to enhanced anti-inflammatory and antioxidant effects. PLGA-FGF21 nanoparticles provide potent protection against APAP-induced ALI by reducing inflammation and oxidative stress, which may be associated with activation of the PGC-1α/Nrf2 signaling pathway, offering a promising strategy for FGF21-based therapy in liver injury.
2026-03-01 | MOTS-c Protects Against Acetaminophen-induced Liver Injury through the MAPK Signaling Pathway.
Acetaminophen (APAP)-induced liver injury (AILI) is a leading cause of acute liver failure worldwide, but effective therapeutic strategies are still lacking. MOTS-c, a mitochondrial-derived peptide, has demonstrated hepatoprotective properties in models of nonalcoholic steatohepatitis (NASH) and hepatitis B virus (HBV) infection. This study aims to explore the role and underlying mechanisms of MOTS-c in AILI. An AILI model was established in male C57BL/6 mice via intraperitoneal (i.p.) injection of APAP (300 mg/kg). The therapeutic potential of MOTS-c and its mechanisms were assessed using behavioral tests, qPCR, western blotting, ELISA, immunohistochemistry, immunofluorescence, and TUNEL staining. MOTS-c levels in both plasma and liver tissues were significantly reduced in APAPinduced AILI mice compared with controls. Administration of MOTS-c via i.p. injection markedly attenuated APAP-induced increases in AST and ALT levels, histopathological liver damage, and other liver injury markers. MOTS-c treatment suppressed the release of pro-inflammatory factors (TNF-α, IL-1β, IL-6, and COX-2) and macrophage infiltration induced by APAP. Furthermore, MOTS-c treatment significantly restored GSH content, diminished reactive oxygen species (ROS) production, and oxidative stress. TUNEL staining confirmed that increased apoptosis in APAPtreated livers was significantly attenuated by MOTS-c, which are key contributors to hepatocyte death and liver injury. Mechanistic studies revealed that MOTS-c inhibited APAP-induced phosphorylation of MAPK pathway components, including ERK, JNK, and p38. The protective effects of MOTS-c on serum ALT and AST levels were abolished by co-treatment with inhibitors of ERK, JNK, and p38. This study reveals that the mitochondrial peptide MOTS-c can alleviate drug-induced liver injury by suppressing oxidative stress and inflammation via the MAPK pathway. This positions MOTS-c as a promising therapeutic candidate for treating APAP-induced liver injury. This study demonstrates that administering MOTS-c effectively protects against APAP-induced liver injury in mice. The protective mechanism involves suppressing the damaging MAPK signaling pathway (ERK, JNK, p38), which in turn reduces oxidative stress, inflammation, and cell death.
2026-02-19 | Annexin A1 enhances liver repair after acetaminophen-induced liver injury by regulating neutrophils function.
Acetaminophen (APAP)-induced acute liver injury (AILI) is a major cause of acute liver failure (ALF), for which effective and timely treatments remain limited. The mechanisms that drive irreversible progression to ALF are not fully understood. Annexin A1 (ANXA1), an anti-inflammatory factor, is implicated in several inflammatory diseases, but its role in APAP-induced liver injury is unclear. In this study, we establish a clinically relevant mouse model of APAP-induced AILI and show that ANXA1 expression increases in the liver, primarily originating from infiltrating neutrophils. ANXA1 deficiency exacerbated liver damage and hindered tissue repair, partly due to the formation of neutrophil extracellular traps (NETs) and altered neutrophil phenotypes. Inhibition of NETs by DNase I restored liver repair and promoted a shift of neutrophils toward an N2 anti-inflammatory phenotype in ANXA1-deficient mice. Furthermore, treatment with Ac2-26, an ANXA1-derived peptide, suppressed NET formation and modulated neutrophil phenotypes to alleviate liver injury. These findings suggest that ANXA1 facilitates liver repair after APAP-induced injury by reprogramming neutrophils via NETs inhibition, offering a promising therapeutic target to enhance recovery from drug-induced liver injury.
other
2025-08-27 | CpG-oligodeoxynucleotides challenged macrophages ameliorate acetaminophen induced liver injury by activating TLR9/IRG1/itaconate metabolic pathway.
Acetaminophen, or N-acetyl-para-aminophenol (APAP), causes severe liver damage and acute liver failure when overdosed. Oligodeoxynucleotides containing CpG motifs (CpG ODN) can regulate the function of macrophages, which play an important role in drug-induced liver injury. It is unclear whether CpG ODN-treated macrophages play an immune regulation role in APAP-induced liver injury. In the present study, we aim to explore the role of CpG ODN-activated macrophages in APAP-induced liver injury and the underlying mechanism in protecting against the cytotoxicity of APAP. In vivo, C57BL/6 mice were treated with APAP (300 mg/Kg) or/and CpG ODN (ODN 1826, 1.65 mg/Kg) by intraperitoneal injection, then survival rate, histopathological evaluation, and inflammatory factors were observed to ascertain the protective effect of CpG ODN. Then, CpG ODN-treated macrophages were reinfused into the animal model to determine the effector cells. In vitro, RNA sequencing and untargeted metabolomics detection were performed to illustrate the underlying mechanism. Last, Acod1 siRNA interference was used to clarify the role of IRG1 in resistance to APAP cytotoxicity by ROS and apoptosis indicator detections. We found that CpG ODN showed a protective effect against APAP cytotoxicity by stimulating macrophages rather than hepatic parenchymal cells. In particular, reinfusion of CpG ODN-treated macrophages to mice can alleviate APAP-induced liver injury. Transcriptome and metabolome analysis revealed that the expression of aconitate decarboxylase 1 (Acod1; also known as immune responsive gene 1, IRG1) and the metabolite itaconate generated by IRG1 catalysis increased after CpG ODN stimulation. In addition, we found that the mechanism of this protective effect is ascribed to the increased expression of Acod1 and the antioxidative function of itaconate by the activation of the TLR9/NF-κB signaling pathway. CpG ODN alleviated liver injury induced by APAP through the activation of the TLR9/NF-κB signaling pathway in macrophages, upregulating the expression of IRG1 protein, promoting the production of endogenous metabolite itaconate, and inhibiting macrophage apoptosis which was regulated by upregulating the expression of Nrf2 to inhibit ROS production. This study sheds new light on CpG ODN as a therapeutic strategy in resistance to APAP-induced liver injury.
2025-07-02 | miR-375 protects against acetaminophen-induced acute liver failure by orchestrating pharmacogene expression.
Acetaminophen (APAP) overdose is a leading cause of acute liver failure (ALF), primarily through the excessive production of N-acetyl-p-benzoquinone imine (NAPQI). N-acetylcysteine (NAC) is the Food and Drug Administration-approved treatment for APAP overdose, but there is a growing interest in microRNAs as potential therapeutic agents. We delivered miR-375 ectopically via a liver-tropic adeno-associated virus serotype 8 (AAV8) and demonstrated its potent protection in a murine model of APAP overdose-induced ALF. Slc16a2, Cyb5b, and Acsl5 were identified as critical targets acting synergistically to mitigate toxicity. Liver transcriptome revealed that miR-375 overexpression or silencing of the targets of miR-375 increased Gstm3 expression in mice. AAV8-mediated Gstm3 expression protects against APAP-ALF, and the protection was further enhanced by disrupting the expression of Cyp2e1. Additionally, CYP2E1 and GSS, which contribute to APAP detoxification, were down- and upregulated by miR-375, respectively. These findings suggest that miR-375 prevents APAP-ALF by orchestrating the expression of pharmacogenes and enhancing glutathione synthesis. We conclude that miR-375 and its targets are promising therapeutic targets for APAP-ALF.
2025-02-28 | Liver-specific Bcl3 Knockout Alleviates Acetaminophen-induced Liver Injury by Activating Nrf2 Pathway in Male Mice.
Acetaminophen (APAP) overdose is the leading cause of acute liver failure, with oxidative stress being a critical factor in this process. Glutathione (GSH) plays a vital defensive role. Activation of nuclear factor erythroid 2 like 2 (Nrf2) pathway mitigates APAP-induced liver damage by promoting GSH biosynthesis and enhancing drug detoxification. Although the role of B cell leukemia/lymphoma 3 (Bcl3) in regulating inflammatory responses, cellular oncogenesis, and immune balance is well-documented, its function in APAP-induced liver injury remains unclear. We employed liver-specific Bcl3 knockout (Bcl3hep-/-) mice and adeno-associated virus (AAV)-8-mediated Bcl3 overexpression (AAV-Bcl3) mice to model APAP-induced liver injury. Liver damage was assessed through hematoxylin and eosin staining and serum alanine aminotransferase and aspartate aminotransferase measurements. The interaction between Bcl3 and Nrf2 was examined using immunofluorescence and co-immunoprecipitation assays. Our study reveals a significant upregulation of Bcl3 expression in the livers of male mice following APAP administration, suggesting Bcl3's potential involvement in this pathological process. In Bcl3hep-/- mice, a reduced severity of liver damage was observed at both 6 and 24 hours post-APAP treatment compared with controls. Notably, Bcl3-deficient mice exhibited accelerated GSH replenishment due to the rapid induction of Gclc and Gclm genes following 6 hours of APAP exposure. Through immunofluorescence and co-immunoprecipitation analyses, we identified an interaction between Bcl3 and Nrf2. The loss of Bcl3 enhanced Nrf2 translocation upon APAP challenge, leading to the upregulation of antioxidant gene expression. These findings suggest that Bcl3 knockout alleviates oxidative stress resulting from APAP overdose. We uncovered a previously uncharacterized role of Bcl3 in APAP-induced liver injury, emphasizing the role of the Bcl3-Nrf2 axis in oxidative stress-related liver damage.
2024-07-28 | Adenosine kinase protects against acetaminophen-induced acute liver injury by activating autophagy in hepatocytes.
Acute liver injury (ALI) is a common life-threatening condition with a high mortality rate due to liver disease-related death. However, current therapeutic interventions for ALI remain ineffective, and the development of effective novel therapies is urgently needed. Liver samples from patients with drug-induced ALI were collected to detect adenosine kinase (ADK) expression. Male C57BL/6 J mice, hepatocyte-specific ADK knockout (ADKHKO) mice, and their controls (ADKf/f) were exposed to acetaminophen (APAP) and other treatments to investigate the mechanisms of APAP-related ALI. ADK expression was significantly decreased in APAP-injured livers. Hepatocyte-specific ADK deficiency exacerbated APAP-induced ALI, while a gain-of-function approach delivering AAV-ADK, markedly alleviated APAP-induced ALI, as indicated by changes in alanine aminotransferases (ALT) levels, aspartate aminotransferase (AST) levels, neutrophil infiltration and hepatocyte death. This study showed that ADK played a critical role in ALI by activating autophagy through two signaling pathways, the adenosine monophosphate-activated protein kinase (AMPK)-mTOR pathway and the adenosine receptor A1 (ADORA1)-Akt-mTOR pathway. Furthermore, we found that metformin upregulated ADK expression in hepatocytes and protected against APAP-induced ALI. These results demonstrate that ADK is critical in protecting against APAP-induced ALI and that developing therapeutics targeting ADK-adenosine-ADORA1 is a new approach for ALI treatment. Metformin is a potential candidate for preventing ALI by upregulating ADK.
2024-03-26 | Deletion of hepatocyte cysteine dioxygenase type 1, a bile acid repressed gene, enhances glutathione synthesis and ameliorates acetaminophen hepatotoxicity.
Liver is a major organ that metabolizes sulfur amino acids cysteine, which is the substrate for the synthesis of many essential cellular molecules including GSH, taurine, and coenzyme A. Bile acid-activated farnesoid x receptor (FXR) inhibits cysteine dioxygenase type 1 (CDO1), which mediates hepatic cysteine catabolism and taurine synthesis. To define the impact of bile acid inhibition of CDO1 on hepatic sulfur amino acid metabolism and antioxidant capacity, we developed hepatocyte-specific CDO1 knockout mice (Hep-CDO1 KO) and hepatocyte specific CDO1 transgenic mice (Hep-CDO1 Tg). Liver metabolomics revealed that genetic deletion of hepatic CDO1 reduced de novo taurine synthesis but had no impact on hepatic taurine abundance or bile acid conjugation. Consistent with reduced cysteine catabolism, Hep-CDO1 KO mice showed increased hepatic cysteine abundance but unaltered methionine cycle intermediates and coenzyme A synthesis. Upon acetaminophen overdose, Hep-CDO1 KO mice showed increased GSH synthesis capacity and alleviated liver injury. In contrast, hepatic CDO1 overexpression in Hep-CDO1 Tg mice stimulated hepatic cysteine to taurine conversion, resulting in reduced hepatic cysteine abundance. However, Hep-CDO1 Tg mice and WT showed similar susceptibility to acetaminophen-induced liver injury. Hep-CDO1 Tg mice showed similar hepatic taurine and coenzyme A compared to WT mice. In summary, these findings suggest that bile acid and FXR signaling inhibition of CDO1-mediated hepatic cysteine catabolism preferentially modulates hepatic GSH synthesis capacity and antioxidant defense, but has minimal effect on hepatic taurine and coenzyme A abundance. Repression of hepatic CDO1 may contribute to the hepatoprotective effects of FXR activation under certain pathologic conditions.
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Drug Discovery Landscape
6 orphan drug designations for Paracetamol poisoning, including 2 approved therapies.
6 orphan drug designations for Paracetamol poisoning, including 2 approved therapies.
Drug | Therapy type | Regulator | Orphan designation | Approval | Sponsor |
|---|---|---|---|---|---|
Fomepizole | small molecules | FDA | 2021-03-02 | — | Denver Health - Rocky Mountain Poison & Drug Safety |
calmangafodipir | small molecules | FDA | 2019-03-15 | — | Egetis Therapeutics AB |
melatonin | small molecules | FDA | 2017-03-02 | — | WORPHMED Srl |
acetylcysteine effervescent tablets for oral solution [Cetylev] | small molecules | FDA | 2015-02-24 | 2016-01-29 | Arbor Pharmaceuticals, Inc. |
acetylcysteine [Acetadote] | small molecules | FDA | 2001-10-19 | 2004-01-23 | Cumberland Pharmaceuticals, Inc. |
Acetylcysteine | small molecules | FDA | 1987-08-13 | — | Bristol-Myers Squibb Company |
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