2. Academic Validation
  3. Hepatocyte-derived extracellular vesicles carrying damaged mitochondria drive neutrophil extracellular traps formation and exacerbate acetaminophen-induced liver injury

Hepatocyte-derived extracellular vesicles carrying damaged mitochondria drive neutrophil extracellular traps formation and exacerbate acetaminophen-induced liver injury

  • Int Immunopharmacol. 2026 Jan 15:169:115944. doi: 10.1016/j.intimp.2025.115944.
Xiaoshuang Jin 1,  Yaning Liu 1,  Peng Chen 1,  Pengxiang Zhao 1,  Yuerong Lu 1,  Haoyu Zhang 1,  Chunran Ma 1,  Zhaosheng Chen 1,  Ge Guan 2,  Guohu Di 3
Affiliations

Affiliations

  • 1 School of Basic Medicine, Qingdao University, 308 Ningxia Road, Qingdao, 266071, China.
  • 2 Organ Transplantation Center, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, China. Electronic address: [email protected].
  • 3 School of Basic Medicine, Qingdao University, 308 Ningxia Road, Qingdao, 266071, China; Institute of Stem Cell and Regenerative Medicine, School of Basic Medicine, Qingdao University, Qingdao, China. Electronic address: [email protected].
PMID: 41349466 DOI: 10.1016/j.intimp.2025.115944
Abstract

Objective: Acetaminophen (APAP) overdose causes severe hepatotoxicity, yet how mitochondrial injury in hepatocytes amplifies innate immune activation remains unclear. This study investigated whether extracellular vesicles released from APAP-injured hepatocytes (APAP-EVs) deliver damaged mitochondrial components to neutrophils, promoting NETs formation and worsening liver injury.

Methods: APAP-induced liver injury was established in C57BL/6 mice. Hepatocyte-derived EVs were isolated and intravenously administered. Liver injury was assessed by ALT, AST, necrosis, Apoptosis, and NETs markers. GW4869, GSK484, RU.521, H-151 and ODN 2088 were used to block EVs release, NETs formation, cGAS-STING or TLR9 signaling.

Results: APAP-EVs treatment markedly exacerbated hepatotoxicity, increasing serum ALT (8775 ± 563.7 U/L vs. 6037 ± 436.5 U/L), AST (8952 ± 670.4 U/L vs. 5539 ± 525.8 U/L), and hepatic necrosis (56.10 ± 1.60 % U/L vs. 30.10 ± 1.52 % U/L) compared with APAP group. Blocking EVs release with GW4869 or inhibiting NETs formation with GSK484 significantly attenuated liver injury. Mechanistically, APAP-EVs activated cGAS-STING signaling in neutrophils to induce NETs formation, an effect abolished by the cGAS inhibitor RU.521 and the STING inhibitor H-151, whereas TLR9 inhibition (ODN 2088) had no effect.

Conclusion: APAP-injured hepatocytes release EVs enriched with damaged mitochondrial cargo that activate cGAS-STING-dependent NETs formation, thereby amplifying liver injury. Targeting EVs biogenesis or NETs formation may provide effective therapeutic strategies for APAP-induced hepatotoxicity.

Keywords

Acetaminophen; Damaged mitochondria; Extracellular vesicles; Neutrophil extracellular traps.

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