Research Paper Volume 16, Issue 19 pp 12697—12725
A new model and precious tool to study molecular mechanisms of macrophage aging
- 1 Sorbonne Université, CNRS UMR 8256 Biological Adaptation and Ageing (B2A), INSERM U1164, Institut de Biologie Paris Seine (IBPS), Paris 75005, France
- 2 INSERM U1148, Laboratory for Vascular and Translational Sciences (LVTS), Université Sorbonne Paris Nord, Bobigny 93000, France
- 3 Sorbonne Université, CNRS, Institut de Biologie Paris Seine (IBPS), Plate-forme Ingénierie des Protéines et Synthèse Peptidique, Paris 75005, France
Received: January 31, 2024 Accepted: August 6, 2024 Published: October 3, 2024
https://doi.org/10.18632/aging.206124How to Cite
Copyright: © 2024 Smith et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
The accumulation of senescent cells, characterized by a senescence-associated secretory phenotype (SASP), contributes to chronic inflammation and age-related diseases (ARD). During aging, macrophages can adopt a senescent-like phenotype and an altered function, which promotes senescent cell accumulation. In the context of aging and ARD, controlling the resolution of the inflammatory response and preventing chronic inflammation, especially by targeting macrophages, must be a priority. Aging being a dynamic process, we developed a model of in vitro murine peritoneal macrophage aging. Our results show that macrophages cultured for 7 or 14 days exhibit a senescence-like phenotype: proliferation decrease, the levels of cyclin-dependent kinase inhibitors p16INK4A and p21CIP1 and of pro-inflammatory SASP components (MCP-1, IL-6, IL-1β, TNF-α, and MMP-9) increase, phagocytosis capacity decline and glycolytic activity is induced. In our model, chronic treatment with CB3, a thioredoxin-1 mimetic anti-inflammatory peptide, completely prevents p21CIP1 increase and enables day 14 macrophages to maintain proliferative activity.
We describe a new model of macrophage aging with a senescence-like phenotype associated with inflammatory, metabolic and functional perturbations. This model is a valuable tool for characterizing macrophage aging mechanisms and developing innovative strategies with promising therapeutical purpose in limiting inflammaging and ARD.
Abbreviations
7-AAD: 7-actinoactinomycin D; ACN: Acetonitrile; ARD: Age-related diseases; C/EBP-β: CCAAT/enhancer binding protein β; CDKI: Cyclin-dependent kinase inhibitors; D0: Day 0; ECAR: Extracellular acidification rate; EdU: 5-Ethynyl-2’-deoxyuridine; FCCP: Carbonyl Cyanide 4-(trifluoromethoxy) phenylhydrazone; Fig: Figure; IL-: Interleukin; KC-GRO: Keratinocyte Chemoattractant-Growth Regulated Oncogene; LPS: Lipopolysaccharide; MALDI-TOF: Matrix-assisted laser desorption/ionization-time of flight; MAPK: Mitogen-Activated Protein Kinase; MCP-1: Monocyte chemoattractant protein 1; M-CSF: Macrophage-Colony Stimulating Factor; MMP-: Matrix metalloproteinase; NADPH: Nicotinamide adenine dinucleotide phosphate; NF-κB: Nuclear factor-kappa B; NLRP3: NOD-like receptor family, pyrin domain containing 3; OCR: Oxygen Consumption Rate; PBS: Phosphate Buffer Salin; RIPA: Radio-immunoprecipitation assay; ROS: Reactive oxygen species; RP-HPLC: Reverse-phase high performance liquid chromatography; RPMI: Roswell Park Memorial Institute medium; RT: Room Temperature; SASP: Senescence-associated secretory phenotype; SA-β-gal: Senescence-associated β-galactosidase; SDS-PAGE: Sodium Dodecyl Sulfate polyacrylamide gel; TCA: Tricarboxylic acid-cycle; TFA: Trifluoroacetic acid; TLR: Toll-like Receptor; TNF-α: Tumor necrosis factor α; Trx-1: Thioredoxin-1; TUNEL: dUTP Nick-End Labeling.