Inflammatory Burden Shapes Antioxidant Adaptation and Immunometabolic Remodeling in LPS-Activated BV-2 Microglial Cells
6 affiliations
- 1Department for Life Quality Studies, Alma Mater Studiorum, University of Bologna, Corso d’Augusto 237, 47921 Rimini, Italy
- 2Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56126 Pisa, Italy
- 3IRCCS Stella Maris Foundation, Viale del Tirreno 331, Calambrone, 56128 Pisa, Italy
- 4Department of Medical, Oral and Biotechnological Sciences, University G. D’Annunzio of Chieti-Pescara, 66100 Chieti, Italy
- 5Department of Biological, Chemical and Pharmaceutical Sciences and Technologies, University of Palermo, Via Archirafi 28, 90123 Palermo, Italy
- 6Department of Pharmacy and Biotechnology, Alma Mater Studiorum, University of Bologna, Via Irnerio, 42, 40126 Bologna, Italy
Abstract
Neuroinflammation is a hallmark of numerous neurodegenerative diseases and is characterized by profound inflammatory, metabolic, and redox alterations in activated microglia. Although lipopolysaccharide (LPS)-stimulated BV-2 cells represent one of the most widely used in vitro models of neuroinflammation, a comprehensive understanding of the coordinated molecular adaptations underlying this model remains incomplete. The aim of this study was to characterize the proteomic remodeling of BV-2 cells exposed to LPS 100 ng/mL for 24 h and investigate the relationship between antioxidant and inflammatory responses across increasing levels of LPS exposure. Proteomic analysis identified 89 differentially expressed proteins involved in cytoskeletal organization, clathrin-mediated endocytosis, lysosomal function, immunometabolism, and redox regulation. Validation experiments confirmed changes in cytoskeletal and endocytic proteins, including Vimentin, Gelsolin, Disabled homolog-2, and clathrin light chain A, together with modulation of proteins involved in the lysosomal pathways and marked induction of the immunometabolic enzyme aconitate decarboxylase 1. Functional enrichment analysis identified Nuclear factor erythroid 2-related factor 2 (NRF2) as the most significantly activated upstream regulator, a prediction consistent with engagement of an adaptive antioxidant program. Interestingly, analysis of increasing LPS concentrations revealed that antioxidant responses were not proportional to the inflammatory stimulus. Expression of NRF2-controlled antioxidant enzymes progressively increased for LPS concentrations up to 500 ng/mL, but declined at 1000 ng/mL LPS, whereas pro-inflammatory mediators remained continuously upregulated. This pattern is consistent with the attenuation of antioxidant adaptation under the highest inflammatory burden tested. Together, these findings demonstrate that the BV-2/LPS model is characterized by a dynamic interplay between inflammatory, redox, metabolic, lysosomal, endocytic, and cytoskeletal pathways, providing a more comprehensive characterization to support the interpretation and experimental use of this neuroinflammatory model.
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BibTeX
@article{barbalace2026inflammatory,
title = {{Inflammatory Burden Shapes Antioxidant Adaptation and Immunometabolic Remodeling in LPS-Activated BV-2 Microglial Cells}},
author = {Maria Cristina Barbalace and Irene Rinaldi and Martina Mantini and Lorenzo Zallocco and Maurizio Ronci and Federica Di Cintio and Alessandro Attanzio and Cecilia Prata and Marco Malaguti and Silvana Hrelia and Laura Giusti and Cristina Angeloni},
journal = {International Journal of Molecular Sciences},
year = {2026},
month = oct,
doi = {10.3390/ijms27208981},
url = {https://www.mdpi.com/1422-0067/27/20/8981},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Inflammatory Burden Shapes Antioxidant Adaptation and Immunometabolic Remodeling in LPS-Activated BV-2 Microglial Cells
AU - Maria Cristina Barbalace
AU - Irene Rinaldi
AU - Martina Mantini
AU - Lorenzo Zallocco
AU - Maurizio Ronci
AU - Federica Di Cintio
AU - Alessandro Attanzio
AU - Cecilia Prata
AU - Marco Malaguti
AU - Silvana Hrelia
AU - Laura Giusti
AU - Cristina Angeloni
PY - 2026
DA - 2026/10/01
JO - International Journal of Molecular Sciences
DO - 10.3390/ijms27208981
UR - https://www.mdpi.com/1422-0067/27/20/8981
AB - Neuroinflammation is a hallmark of numerous neurodegenerative diseases and is characterized by profound inflammatory, metabolic, and redox alterations in activated microglia. Although lipopolysaccharide (LPS)-stimulated BV-2 cells represent one of the most widely used in vitro models of neuroinflammation, a comprehensive understanding of the coordinated molecular adaptations underlying this model remains incomplete. The aim of this study was to characterize the proteomic remodeling of BV-2 cells exposed to LPS 100 ng/mL for 24 h and investigate the relationship between antioxidant and inflammatory responses across increasing levels of LPS exposure. Proteomic analysis identified 89 differentially expressed proteins involved in cytoskeletal organization, clathrin-mediated endocytosis, lysosomal function, immunometabolism, and redox regulation. Validation experiments confirmed changes in cytoskeletal and endocytic proteins, including Vimentin, Gelsolin, Disabled homolog-2, and clathrin light chain A, together with modulation of proteins involved in the lysosomal pathways and marked induction of the immunometabolic enzyme aconitate decarboxylase 1. Functional enrichment analysis identified Nuclear factor erythroid 2-related factor 2 (NRF2) as the most significantly activated upstream regulator, a prediction consistent with engagement of an adaptive antioxidant program. Interestingly, analysis of increasing LPS concentrations revealed that antioxidant responses were not proportional to the inflammatory stimulus. Expression of NRF2-controlled antioxidant enzymes progressively increased for LPS concentrations up to 500 ng/mL, but declined at 1000 ng/mL LPS, whereas pro-inflammatory mediators remained continuously upregulated. This pattern is consistent with the attenuation of antioxidant adaptation under the highest inflammatory burden tested. Together, these findings demonstrate that the BV-2/LPS model is characterized by a dynamic interplay between inflammatory, redox, metabolic, lysosomal, endocytic, and cytoskeletal pathways, providing a more comprehensive characterization to support the interpretation and experimental use of this neuroinflammatory model.
SN - 1422-0067
ER -