p65 enhanced METTL3-mediated m6A methylation of HMGB1 to promote microglia M1 polarization in sepsis-associated encephalopathy

Xiaowen Wu; Jie Wang; Jun Li; Guosheng Yao; Linyun Wei; Xuebin Li

doi:10.5114/ceji.2025.154882

eISSN: 1644-4124
ISSN: 1426-3912

Central European Journal of Immunology

Current issue Archive Manuscripts accepted About the journal Special Issues Editorial board Abstracting and indexing Subscription Contact Instructions for authors Publication charge

Editorial System

Submit your Manuscript

Send email

Copy url:

abstract:

Original paper

p65 enhanced METTL3-mediated m6A methylation of HMGB1 to promote microglia M1 polarization in sepsis-associated encephalopathy

Xiaowen Wu

¹

,

Jie Wang

²

,

Jun Li

¹

,

Guosheng Yao

¹

,

Linyun Wei

¹

,

Xuebin Li

^{3, 4, 5}

Department of Intensive Care Medicine, The Affiliated Hospital of Youjiang Medical University for Nationalities, Baise 533000, Guangxi, China
Department of Nephrology, The Affiliated Hospital of Youjiang Medical University for Nationalities, Baise 533000, Guangxi, China
Department of Neurology, The Affiliated Hospital of Youjiang Medical University for Nationalities, Baise 533000, Guangxi, China
School of Clinical Medicine, Youjiang Medical University for Nationalities, Baise 533000, Guangxi, China
Biological Molecule Laboratory, Guangxi University Key Laboratory of High Incidence Prevention and Control Research in Western Guangxi, Baise, Guangxi, China

Cent Eur J Immunol 2025; 50 (4)

DOI: https://doi.org/10.5114/ceji.2025.154882

Online publish date: 2025/10/21

View full text Get citation

PlumX metrics:

Introduction
Sepsis-associated encephalopathy (SAE) is a complication posing a significant risk to patient health and survival. Microglial polarization and inflammation are key to the pathological progression of SAE. The p65 subunit is a component of the nuclear factor-κB (NF-κB) family. This study aimed to clarify the role of p65 in microglial polarization.

Material and methods
Human microglial HMC3 cells were treated with lipopolysaccharide (LPS). Cell viability was measured using a CCK-8 kit. Proinflammatory cytokine levels were evaluated using an ELISA assay. qRT-PCR or western blot assays were used to estimate mRNA or protein levels. The proportion of microglial M1 polarization was assessed using flow cytometry. The m6A-modified high mobility group box 1 (HMGB1) levels were analyzed using MeRIP-qPCR. Dual-luciferase reporter, chromatin immunoprecipitation (ChIP), and RNA immunoprecipitation (RIP) assays were conducted to validate the interactions between p65 and methyltransferase-like 3 (METTL3) or between METTL3 and HMGB1.

Results
LPS promoted microglial M1 polarization and enhanced p65, METTL3, and HMGB1 expression. Further, the inhibition of p65 ameliorated the LPS-induced M1 microglial polarization. p65 promotes HMGB1 m6A methylation by transcriptionally activating METTL3. Under LPS treatment, p65 enhances microglial M1 polarization via METTL3 activation. METTL3 aggravates LPS-induced microglial M1 polarization by positively regulating HMGB1 m6A modification.

Conclusions
p65 increases LPS-induced M1 polarization by promoting METTL3-mediated m6A modification of HMGB1 in HMC3 cells.

keywords:

p65 enhanced METTL3-mediated m6A methylation of HMGB1 to promote microglia M1 polarization in sepsis-associated encephalopathy

Xiaowen Wu 1 , Jie Wang 2 , Jun Li 1 , Guosheng Yao 1 , Linyun Wei 1 , Xuebin Li 3, 4, 5

SAE, p65, METTL3, HMGB1, microglia polarization

Xiaowen Wu

¹

,

Jie Wang

²

,

Jun Li

¹

,

Guosheng Yao

¹

,

Linyun Wei

¹

,

Xuebin Li

^{3, 4, 5}