ORIGINAL PAPER
The AGO2–ATOX1 axis exacerbates inflammation in a mouse sepsis model
1
Department of Emergency Medicine, The Affiliated Nanhua Hospital, Hengyang Medical School, University of South China, Hengyang 421002, Hunan, China
2
Department of Laboratory Medicine, Hengyang First People’s Hospital, Hengyang 421002, Hunan, China
Submission date: 2025-04-10
Final revision date: 2025-07-11
Acceptance date: 2025-08-08
Online publication date: 2026-04-27
Corresponding author
Lipu Deng
Department of Emergency Medicine, The Affiliated Nanhua Hospital, Hengyang Medical School, University of South China, Hengyang 421002, Hunan, China
Department of Emergency Medicine, The Affiliated Nanhua Hospital, Hengyang Medical School, University of South China, Hengyang 421002, Hunan, China
KEYWORDS
ABSTRACT
Introduction:
The study aimed to investigate the mechanism of the Argonaute RISC catalytic component 2 (AGO2)–antioxidant protein 1 (ATOX1) axis in the inflammatory response of sepsis.
Material and Methods:
AGO2 was knocked down by tail vein injection of sh-Ago2 lentivirus, and a sepsis mouse model was established using the cecal ligation and puncture (CLP) method. RAW264.7 cells were transfected with AGO2 or ATOX1 knockdown or overexpression plasmids and treated with lipopolysaccharide (LPS) to construct a cellular sepsis model. Hematoxylin-eosin staining was used to evaluate lung and liver tissue damage in mice. Serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured using biochemical analysis. ELISA was performed to determine the levels of inflammatory cytokines: tumor necrosis factor a (TNF-a), interleukin (IL)-6, and IL-1B. The expression of AGO2, ATOX1, inducible nitric oxide synthase (iNOS), and arginase 1 (Arg1) was analyzed using RT-qPCR and Western blotting. The effect of AGO2 on ATOX1 mRNA stability was assessed using an actinomycin D assay, and the interaction between AGO2 and ATOX1 was analyzed via RIP-qPCR.
Results:
AGO2 and ATOX1 were highly expressed in septic mice. In vitro experiments demonstrated that knockdown of AGO2 or ATOX1 suppressed LPS-induced inflammatory responses and macrophage polarization imbalance. AGO2 promoted M1 macrophage polarization and aggravated inflammatory damage by enhancing the stability of ATOX1 mRNA. In vivo experiments further confirmed that AGO2 knockdown significantly alleviated sepsis-induced lung and liver damage, reduced the inflammatory response, and inhibited ATOX1 expression.
Conclusions:
AGO2 binds to ATOX1 mRNA, promoting ATOX1 expression, inducing macrophage polarization imbalance, and exacerbating sepsis-induced inflammatory responses.
The study aimed to investigate the mechanism of the Argonaute RISC catalytic component 2 (AGO2)–antioxidant protein 1 (ATOX1) axis in the inflammatory response of sepsis.
Material and Methods:
AGO2 was knocked down by tail vein injection of sh-Ago2 lentivirus, and a sepsis mouse model was established using the cecal ligation and puncture (CLP) method. RAW264.7 cells were transfected with AGO2 or ATOX1 knockdown or overexpression plasmids and treated with lipopolysaccharide (LPS) to construct a cellular sepsis model. Hematoxylin-eosin staining was used to evaluate lung and liver tissue damage in mice. Serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured using biochemical analysis. ELISA was performed to determine the levels of inflammatory cytokines: tumor necrosis factor a (TNF-a), interleukin (IL)-6, and IL-1B. The expression of AGO2, ATOX1, inducible nitric oxide synthase (iNOS), and arginase 1 (Arg1) was analyzed using RT-qPCR and Western blotting. The effect of AGO2 on ATOX1 mRNA stability was assessed using an actinomycin D assay, and the interaction between AGO2 and ATOX1 was analyzed via RIP-qPCR.
Results:
AGO2 and ATOX1 were highly expressed in septic mice. In vitro experiments demonstrated that knockdown of AGO2 or ATOX1 suppressed LPS-induced inflammatory responses and macrophage polarization imbalance. AGO2 promoted M1 macrophage polarization and aggravated inflammatory damage by enhancing the stability of ATOX1 mRNA. In vivo experiments further confirmed that AGO2 knockdown significantly alleviated sepsis-induced lung and liver damage, reduced the inflammatory response, and inhibited ATOX1 expression.
Conclusions:
AGO2 binds to ATOX1 mRNA, promoting ATOX1 expression, inducing macrophage polarization imbalance, and exacerbating sepsis-induced inflammatory responses.
REFERENCES (35)
1.
Hawiger J, Veach RA, Zienkiewicz J (2015): New paradigms in sepsis: from prevention to protection of failing microcirculation. J Thromb Haemost 13: 1743-1756.
2.
Salomão R, Ferreira BL, Salomão MC, et al. (2019): Sepsis: evolving concepts and challenges. Braz J Med Biol Res 52: e8595.
3.
Nedeva C (2021): Inflammation and cell death of the innate and adaptive immune system during sepsis. Biomolecules 11: 1011.
4.
De Freitas Caires N, Gaudet A, Portier L, et al. (2018): Endocan, sepsis, pneumonia, and acute respiratory distress syndrome. Crit Care 22: 280.
5.
Gotts JE, Matthay MA (2016): Sepsis: pathophysiology and clinical management. BMJ 353: i1585.
6.
Rudd KE, Johnson SC, Agesa KM, et al. (2020): Global, regional, and national sepsis incidence and mortality, 1990-2017: analysis for the Global Burden of Disease Study. Lancet 395: 200-211.
7.
Fowler AA, 3rd, Truwit JD, Hite RD, et al. (2019): Effect of vitamin C infusion on organ failure and biomarkers of inflammation and vascular injury in patients with sepsis and severe acute respiratory failure: The CITRIS-ALI Randomized Clinical Trial. JAMA 322: 1261-1270.
8.
Yunna C, Mengru H, Lei W, et al. (2020): Macrophage M1/M2 polarization. Eur J Pharmacol 877: 173090.
9.
Butt Y, Kurdowska A, Allen TC (2016): Acute lung injury: A cli-nical and molecular review. Arch Pathol Lab Med 140: 345-350.
10.
Chen X, Liu Y, Gao Y, et al. (2021): The roles of macrophage polarization in the host immune response to sepsis. Int Immunopharmacol 96: 107791.
11.
Lobo V, Nowak I, Fernandez C, et al. (2024): Loss of Lamin A leads to the nuclear translocation of AGO2 and compromised RNA interference. Nucleic Acids Res 52: 9917-9935.
12.
Lessel D, Zeitler DM, Reijnders MRF, et al. (2020): Germline AGO2 mutations impair RNA interference and human neurological development. Nat Commun 11: 5797.
13.
Machado-Pereira M, Saraiva C, Bernardino L, et al. (2022): Argonaute-2 protects the neurovascular unit from damage caused by systemic inflammation. J Neuroinflammation 19: 11.
14.
Zou Z, Lin Q, Yang H, et al. (2020): Nrp-1 mediated plasmatic Ago2 binding miR-21a-3p internalization: A novel mechanism for miR-21a-3p accumulation in renal tubular epithelial cells during sepsis. Biomed Res Int 2020: 2370253.
15.
Bam M, Yang X, Zumbrun EE, et al. (2017): Decreased AGO2 and DCR1 in PBMCs from War Veterans with PTSD leads to diminished miRNA resulting in elevated inflammation. Transl Psychiatry 7: e1222.
16.
Maung MT, Carlson A, Olea-Flores M, et al. (2021): The molecular and cellular basis of copper dysregulation and its relationship with human pathologies. FASEB J 35: e21810.
17.
Das A, Sudhahar V, Ushio-Fukai M, et al. (2019): Novel interaction of antioxidant-1 with TRAF4: role in inflammatory responses in endothelial cells. Am J Physiol Cell Physiol 317: C1161-C1171.
18.
Xu D, Zhu W, Ding C, et al. (2023): Self-homeostasis immunoregulatory strategy for implant-related infections through remodeling redox balance. ACS Nano 17: 4574-4590.
19.
Chen M, Chen Y, Fu R, et al. (2024): Atox1 regulates macrophage polarization in intestinal inflammation via ROS-NLRP3 inflammasome pathway. J Transl Med 22: 497.
20.
Li C, Yu L, Mai C, et al. (2021): KLF4 down-regulation resulting from TLR4 promotion of ERK1/2 phosphorylation underpins inflammatory response in sepsis. J Cell Mol Med 25: 2013-2024.
21.
Delano MJ, Ward PA (2016): The immune system’s role in sepsis progression, resolution, and long-term outcome. Immunol Rev 274: 330-353.
22.
Ma S, Evans R G, Iguchi N, et al. (2019): Sepsis-induced acute kidney injury: A disease of the microcirculation. Microcirculation 26: e12483.
23.
Ehlting C, Wolf SD, Bode JG (2021): Acute-phase protein synthesis: a key feature of innate immune functions of the liver. Biol Chem 402: 1129-1145.
24.
Zhang K, Pomyen Y, Barry AE, et al. (2020): AGO2 mediates MYC mRNA stability in hepatocellular carcinoma. Mol Cancer Res 18: 612-622.
25.
Arroyo JD, Chevillet JR, Kroh EM, et al. (2011): Argonaute2 complexes carry a population of circulating microRNAs independent of vesicles in human plasma. Proc Natl Acad Sci U S A 108: 5003-5008.
26.
Guo Z, Geller DA (2014): microRNA and human inducible nitric oxide synthase. Vitam Horm 96: 19-27.
27.
Mazumder A, Bose M, Chakraborty A, et al. (2013): A transient reversal of miRNA-mediated repression controls macrophage activation. EMBO Rep 14: 1008-1016.
28.
De Kerckhove M, Tanaka K, Umehara T, et al. (2018): Targeting miR-223 in neutrophils enhances the clearance of Staphylococcus aureus in infected wounds. EMBO Mol Med 10: e9024.
29.
Wan L, Jia RM, Ji LL, et al. (2022): AMPK-autophagy-mediated inhibition of microRNA-30a-5p alleviates morphine tolerance via SOCS3-dependent neuroinflammation suppression. J Neuroinflammation 19: 25.
30.
Yu H, Qin L, Peng Y, et al. (2020): Exosomes derived from hypertrophic cardiomyocytes induce inflammation in macrophages via miR-155 mediated MAPK pathway. Front Immunol 11: 606045.
31.
Sudhahar V, Shi Y, Kaplan JH, et al. (2022): Whole-transcriptome sequencing analyses of nuclear antixoxidant-1 in endothelial cells: role in inflammation and atherosclerosis. Cells 11: 2919.
32.
Zou M, Zhang W, Zhu Y, et al. (2023): Identification of 6 cuproptosis-related genes for active ulcerative colitis with both diagnostic and therapeutic values. Medicine (Baltimore) 102: e35503.
33.
Kim DW, Shin MJ, Choi YJ, et al. (2018): Tat-ATOX1 inhibits inflammatory responses via regulation of MAPK and NF-B pathways. BMB Rep 51: 654-659.
34.
Wang Y, Chen Z, Liang K, et al. (2025): AGO2 mediates immunotherapy failure via suppressing tumor IFN-gamma response-dependent CD8(+) T cell immunity. Cell Rep 44: 115445.
35.
Yu H, Si G, Si F (2024): Mendelian randomization validates the immune landscape mediated by aggrephagy in esophageal squamous cell carcinoma patients from the perspectives of multi-omics. J Cancer 15: 1940-1953.
Share
RELATED ARTICLE
| eISSN: | 1644-4124 |
| ISSN: | 1426-3912 |
We process personal data collected when visiting the website. The function of obtaining information about users and their behavior is carried out by voluntarily entered information in forms and saving cookies in end devices. Data, including cookies, are used to provide services, improve the user experience and to analyze the traffic in accordance with the Privacy policy. Data are also collected and processed by Google Analytics tool (more).
You can change cookies settings in your browser. Restricted use of cookies in the browser configuration may affect some functionalities of the website.
You can change cookies settings in your browser. Restricted use of cookies in the browser configuration may affect some functionalities of the website.
