2. Academic Validation
  3. CpG-oligodeoxynucleotides challenged macrophages ameliorate acetaminophen induced liver injury by activating TLR9/IRG1/itaconate metabolic pathway

CpG-oligodeoxynucleotides challenged macrophages ameliorate acetaminophen induced liver injury by activating TLR9/IRG1/itaconate metabolic pathway

  • Mol Med. 2025 Aug 25;31(1):282. doi: 10.1186/s10020-025-01324-0.
Yibai Qu # 1 2,  Zehui Jiang # 3,  Zhixia Chen # 4,  Sidan Luo # 5 3,  Bingyao Xie 3,  Xubo Wu 6,  Gang Yuan 3,  Kan Wu 3,  Li Chen 1,  Tian Tian 3,  Shan Li 3,  Haihua Luo 3,  Quan Li 4,  Ding Yuan 2,  Yan Zhang 2,  Yanxia Gao 7,  Jun Zhou 8,  Zhengzheng Yan 9 10,  Yong Jiang 11 12 13 14
Affiliations

Affiliations

  • 1 Department of Anesthesiology, The Third Affiliated Hospital of Southern Medical University, Guangzhou, 510630, China.
  • 2 Department of Emergency Medicine, Henan Provincial Medical Key Laboratory of Toxicological Diseases, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450001, China.
  • 3 Guangdong Provincial Key Laboratory of Proteomics, School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, China.
  • 4 Department of Anesthesiology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital and Shenzhen Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Shenzhen, 518116, China.
  • 5 Department of Anesthesiology, The Tenth Affiliated Hospital (Dongguan People's Hospital), Southern Medical University, Dongguan, Guangdong, 523059, China.
  • 6 Department of Hepatobiliary and Pancreatic Surgery, Minhang Hospital, Fudan University, Shanghai, 201199, China.
  • 7 Department of Emergency Medicine, Henan Provincial Medical Key Laboratory of Toxicological Diseases, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450001, China. gaoyanxiazzu@163.com.
  • 8 Department of Anesthesiology, The Third Affiliated Hospital of Southern Medical University, Guangzhou, 510630, China. zhoujun7843@126.com.
  • 9 Department of Anesthesiology, The Tenth Affiliated Hospital (Dongguan People's Hospital), Southern Medical University, Dongguan, Guangdong, 523059, China. 470575280@qq.com.
  • 10 Dongguan Key Laboratory of Anesthesia and Organ Protection, Dongguan, 523059, Guangdong, China. 470575280@qq.com.
  • 11 Department of Anesthesiology, The Third Affiliated Hospital of Southern Medical University, Guangzhou, 510630, China. jiang48231@163.com.
  • 12 Guangdong Provincial Key Laboratory of Proteomics, School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, China. jiang48231@163.com.
  • 13 State Key Laboratory of Metabolic Dysregulation & Prevention and Treatment of Esophageal Cancer, Henan Key Laboratory of Critical Care Medicine, Henan International Joint Laboratory of Infection and Immunology, Department of Emergency Medicine, The First Affiliated Hospital, Zhengzhou University, Zhengzhou, 450001, China. jiang48231@163.com.
  • 14 Institute of Infection and Immunity, Henan Academy of Innovations in Medical Science, Zhengzhou, China. jiang48231@163.com.
  • # Contributed equally.
PMID: 40855408 DOI: 10.1186/s10020-025-01324-0
Abstract

Background: 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.

Methods: 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.

Results: 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.

Conclusion: 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.

Keywords

Acetaminophen; IRG1; Itaconate; Liver injury; Macrophage apoptosis; Mitochondrial dysfunction.

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