ORIGINAL PAPER
Steatotic hepatocyte-derived extracellular vesicle miR-192 modulates M2-associated marker expression by suppressing IL6ST
1
Department of Hematology, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Southern Medical University, Guangdong, China
2
Department of Immunology, West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University, Chengdu, China
Submission date: 2026-03-12
Final revision date: 2026-07-29
Acceptance date: 2026-08-18
Online publication date: 2026-10-30
Corresponding author
Miao Wu
Department of Hematology, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Southern Medical University, Guangdong, China
Department of Hematology, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Southern Medical University, Guangdong, China
KEYWORDS
ABSTRACT
Introduction:
Extracellular vesicles, which transport microRNAs, are emerging biomarkers for metabolic dysfunction-associated steatotic liver disease. Although extracellular vesicles modulate nearby hepatocytes, their influence on macrophages – central to disease progression – remains unclear. We aimed to identify functional microRNAs in extracellular vesicles and elucidate their role in macrophage activation and polarization.
Material and Methods:
Extracellular vesicles were isolated from steatotic AML12 hepatocytes using ultracentrifugation and characterized by transmission electron microscopy. Cellular uptake was confirmed using PKH67 labeling and confocal microscopy. RAW264.7 macrophages were treated with extracellular vesicles or microRNA mimics. Predicted microRNA targets were validated using a dual luciferase assay and quantitative real-time polymerase chain reaction (PCR). Macrophage polarization markers (M1/M2) were analyzed using quantitative real-time PCR. Statistical significance was set at p < 0.05 (GraphPad Prism).
Results:
Extracellular vesicles from steatotic AML12 hepatocytes showed enrichment of microRNAs (e.g., mmu-miR-122, -21, and -34a), with mmu-miR-192 showing the greatest increase (2.2-fold; p < 0.01). These extracellular vesicles were taken up by macrophages. Treatment with extracellular vesicle-derived miR-192 suppressed IL6ST expression (p < 0.05) and inhibited expression of M1 markers (e.g., iNOS, TNF-a, and IL-1b), while enhancing the expression of M2 markers (e.g., CD206 and IL-10) (p < 0.05). Restoring IL6ST expression reversed these effects.
Conclusions:
Our findings suggest that extracellular vesicles may mediate miR-192 transfer from steatotic hepatocytes to macrophages in metabolic dysfunction-associated steatotic liver disease. By targeting IL6ST and shifting macrophage marker expression toward an M2-associated profile, the miR-192/IL6ST axis may contribute to the regulation of steatotic liver inflammation and warrants further investigation as a potential therapeutic target.
Extracellular vesicles, which transport microRNAs, are emerging biomarkers for metabolic dysfunction-associated steatotic liver disease. Although extracellular vesicles modulate nearby hepatocytes, their influence on macrophages – central to disease progression – remains unclear. We aimed to identify functional microRNAs in extracellular vesicles and elucidate their role in macrophage activation and polarization.
Material and Methods:
Extracellular vesicles were isolated from steatotic AML12 hepatocytes using ultracentrifugation and characterized by transmission electron microscopy. Cellular uptake was confirmed using PKH67 labeling and confocal microscopy. RAW264.7 macrophages were treated with extracellular vesicles or microRNA mimics. Predicted microRNA targets were validated using a dual luciferase assay and quantitative real-time polymerase chain reaction (PCR). Macrophage polarization markers (M1/M2) were analyzed using quantitative real-time PCR. Statistical significance was set at p < 0.05 (GraphPad Prism).
Results:
Extracellular vesicles from steatotic AML12 hepatocytes showed enrichment of microRNAs (e.g., mmu-miR-122, -21, and -34a), with mmu-miR-192 showing the greatest increase (2.2-fold; p < 0.01). These extracellular vesicles were taken up by macrophages. Treatment with extracellular vesicle-derived miR-192 suppressed IL6ST expression (p < 0.05) and inhibited expression of M1 markers (e.g., iNOS, TNF-a, and IL-1b), while enhancing the expression of M2 markers (e.g., CD206 and IL-10) (p < 0.05). Restoring IL6ST expression reversed these effects.
Conclusions:
Our findings suggest that extracellular vesicles may mediate miR-192 transfer from steatotic hepatocytes to macrophages in metabolic dysfunction-associated steatotic liver disease. By targeting IL6ST and shifting macrophage marker expression toward an M2-associated profile, the miR-192/IL6ST axis may contribute to the regulation of steatotic liver inflammation and warrants further investigation as a potential therapeutic target.
REFERENCES (26)
1.
Chan WK, Chuah KH, Rajaram RB, Lim LL, et al. (2023): Metabolic dysfunction-associated steatotic liver disease (MASLD): A state-of-the-art review. J Obes Metab Syndr 32: 197-213.
2.
Do A, Zahrawi F, Mehal WZ (2025): Therapeutic landscape of metabolic dysfunction-associated steatohepatitis (MASH). Nat Rev Drug Discov 24: 171-189.
3.
Tincopa MA, Anstee QM, Loomba R (2024): New and emerging treatments for metabolic dysfunction-associated steatohepatitis. Cell Metab 36: 912-926.
4.
Castera L, Friedrich-Rust M, Loomba R (2019): Noninvasive assessment of liver disease in patients with nonalcoholic fatty liver disease. Gastroenterology 156: 1264-1281.e4.
5.
European Association for the Study of the Liver (EASL); European Association for the Study of Diabetes (EASD); European Association for the Study of Obesity (EASO) (2024): EASL-EASD-EASO Clinical Practice Guidelines on the management of metabolic dysfunction-associated steatotic liver disease (MASLD). J Hepatol 81: 492-542.
6.
Abdelhameed F, Kite C, Lagojda L, Dallaway A, et al. (2024): Non-invasive scores and serum biomarkers for fatty liver in the era of metabolic dysfunction-associated steatotic liver disease (MASLD): A comprehensive review from NAFLD to MAFLD and MASLD. Curr Obes Rep 13: 510-531.
7.
Mily A, Kalsum S, Loreti MG, Rekha RS, et al. (2020): Polarization of M1 and M2 human monocyte-derived cells and analysis with flow cytometry upon Mycobacterium tuberculosis infection. J Vis Exp 163: 61807.
8.
Wang Z, Du K, Jin N, Tang B, et al. (2023): Macrophage in liver fibrosis: identities and mechanisms. Int Immunopharmacol 120: 110357.
9.
Guo W, Li Z, Anagnostopoulos G, Kong WT, et al. (2024): Notch signaling regulates macrophage-mediated inflammation in metabolic dysfunction-associated steatotic liver disease. Immunity 57: 2310-2327.e6.
10.
Shaker O, Mahfouz H, Salama A, Medhat E (2020): Long non-coding HULC and miRNA-372 as diagnostic biomarkers in hepatocellular carcinoma. Rep Biochem Mol Biol 9: 230-240.
11.
Yu J, Peng J, Luan Z, Zheng F, et al. (2019): MicroRNAs as a novel tool in the diagnosis of liver lipid dysregulation and fatty liver disease. Molecules 24: 230.
12.
Saad El-Din S, Ahmed Rashed L, Eissa M, Eldemery AB, et al. (2022): Potential role of circRNA-HIPK3/microRNA-124a crosstalk in the pathogenesis of rheumatoid arthritis. Rep Biochem Mol Biol 10: 527-536.
13.
Carpi S, Daniele S, de Almeida JFM, Gabbia D (2024): Recent advances in miRNA-based therapy for MASLD/MASH and MASH-associated HCC. Int J Mol Sci 25: 12229.
14.
Vishnoi A, Rani S (2023): miRNA biogenesis and regulation of diseases: An updated overview. Methods Mol Biol 2595: 1-12.
15.
Harrell CR, Jovicic N, Djonov V, Arsenijevic N, et al. (2019): Mesenchymal stem cell-derived exosomes and other extracellular vesicles as new remedies in the therapy of inflammatory diseases. Cells 8: 1605.
16.
Stefan N, Yki-Järvinen H, Neuschwander-Tetri BA (2025): Metabolic dysfunction-associated steatotic liver disease: heterogeneous pathomechanisms and effectiveness of metabolism-based treatment. Lancet Diabetes Endocrinol 13: 134-148.
17.
Ye J, Zheng J, Tian X, Xu B, et al. (2022): Fucoxanthin attenuates free fatty acid-induced nonalcoholic fatty liver disease by regulating lipid metabolism/oxidative stress/inflammation via the AMPK/Nrf2/TLR4 signaling pathway. Mar Drugs 20: 225.
18.
Baxter AA, Phan TK, Hanssen E, Liem M, et al. (2019): Analysis of extracellular vesicles generated from monocytes under conditions of lytic cell death. Sci Rep 9: 7538.
19.
Zhong Y, Qin C, Wang Q, Ding M, et al. (2024): Inhibition of Foxp3 expression in the placenta of mice infected intraperitoneally by toxoplasma gondii tachyzoites: insights into the PPAR/miR-7b-5p/Sp1 signaling pathway. Parasit Vectors 17: 189.
20.
Liang CL, Jiang H, Feng W, Liu H, et al. (2021): Total glucosides of paeony ameliorate pristane-induced lupus nephritis by inducing PD-1 ligands+ macrophages via activating IL-4/STAT6/PD-L2 signaling. Front Immunol 12: 683249.
21.
Zhang Y, Li X, Wang C, Zhang M, et al. (2020): lncRNA AK085865 promotes macrophage M2 polarization in CVB3-induced VM by regulating ILF2-ILF3 complex-mediated miRNA-192 biogenesis. Mol Ther Nucleic Acids 21: 441-451.
22.
Essandoh K, Li Y, Huo J, Fan GC (2016): MiRNA-mediated macrophage polarization and its potential role in the regulation of inflammatory response. Shock 46: 122-131.
23.
Kang H, Liu T, Wang Y, Bai W, et al. (2024): Neutrophil-macrophage communication via extracellular vesicle transfer promotes itaconate accumulation and ameliorates cytokine storm syndrome. Cell Mol Immunol 21: 689-706.
24.
Abdelgwad M, Zakaria R, Marzouk S, Sabry D, et al. (2023): The emerging role of circular RNA homeodomain interacting protein kinase 3 and circular RNA 0046367 through Wnt/beta-catenin pathway on the pathogenesis of nonalcoholic steatohepatitis in Egyptian patients. Rep Biochem Mol Biol 11: 614-625.
25.
Liu B, Li J, Chen B, Shuai Y, et al. (2023): Dental pulp stem cells induce anti-inflammatory phenotypic transformation of macrophages to enhance osteogenic potential via IL-6/GP130/STAT3 signaling. Ann Transl Med 11: 90.
26.
Yao Y, Chen C, Wang J, Xuan H, et al. (2023): Circular RNA circATP9A promotes non-small cell lung cancer progression by interacting with HuR and by promoting extracellular vesicles-mediated macrophage M2 polarization. J Exp Clin Cancer Res 42: 330.
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.
