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Updated: May 14, 2025
Investigation of Macrophage Polarization Using Bone Marrow Derived Macrophages
Published on: June 23, 2013
KLF15 regulates macrophage polarization patterns in deep vein thrombosis
Jizheng Li1, YaoYu Xiang2, Haopeng Shentu2
1The First Clinical Medical College, The Yunnan University of Chinese Medicine, No.1076 Yuhua Road, Kunming 650500, PR China.
Background:
KLF15 is involved in cardiovascular disease processes by regulating vascular remodeling and metabolic disorders. Macrophages mediate the inflammatory response in deep vein thrombosis (DVT) by secreting inflammatory cytokines and modulating the fibrinolytic system. Therefore, this study aims to discuss the effect of KLF15 on macrophage polarization in DVT.
Methods:
In vivo, a DVT animal model was used to assess KLF15 expression and macrophage polarization. In vitro, PMA-treated THP-1 cells with KLF15 overexpression were differentiated into M1- and M2-like macrophages, with polarization markers analyzed by molecular and cellular assays.
Results:
In vivo experiments, levels of KLF15, iNOS, CD206, IL-1β, IL-6, IL-10 and TGF-β were increased in the DVT animal model. In vitro experiments, KLF15 overexpression augmented iNOS, CD86, IL-12, TNF-α, IL-1β, and IL-6 levels in M1-like macrophages. Additionally, KLF15 overexpression diminished CD206, IL-10, ARG1, IL-10 and TGF-β levels in M2-like macrophages. In CUT&Tag, peaks bound to KLF15 and lgG were mainly located in the promoter and intronic regions, and KLF15 protein bound more peaks than lgG near the TSS site. YY1, EIF4E, LCK, HMGB1, GPD2, MORF4L1, HIPK2 and NEK2 were hub genes in the peaks that bind to KLF15. ChIP assay confirmed that KLF15 bound the NEK2 promoter in M1-like macrophages and enhanced its transcription and NF-κB pathway activity.
Conclusion:
KLF15 facilitates M1 macrophage polarization in DVT via the NEK2/NF-κB pathway, highlighting its potential as a therapeutic target for DVT management. Future studies are warranted to explore its clinical applicability and mechanistic nuances.
Insights
Krüppel-like factor 15 (KLF15) promotes M1 macrophage polarization in deep vein thrombosis (DVT) through the NEK2/NF-κB pathway. This finding suggests KLF15 as a potential therapeutic target for managing DVT.
Area of Science:
- Cardiovascular Biology
- Immunology
- Molecular Biology
Background:
- Krüppel-like factor 15 (KLF15) is implicated in cardiovascular diseases, influencing vascular remodeling and metabolic functions.
- Macrophages play a critical role in deep vein thrombosis (DVT) by mediating inflammation and modulating the fibrinolytic system.
Purpose of the Study:
- To investigate the role of KLF15 in macrophage polarization within the context of DVT.
- To elucidate the molecular mechanisms by which KLF15 affects macrophage phenotypes in DVT.
Main Methods:
- Establishment of a DVT animal model to analyze KLF15 expression and macrophage polarization in vivo.
- In vitro studies using PMA-treated THP-1 cells with KLF15 overexpression to induce M1 and M2-like macrophage polarization, followed by molecular and cellular assays.
- Chromatin accessibility assays (CUT&Tag) and Chromatin immunoprecipitation (ChIP) to identify KLF15 binding sites and downstream targets.
Main Results:
- In vivo, DVT models showed elevated levels of KLF15 and inflammatory markers (iNOS, IL-1β, IL-6) alongside altered macrophage markers (CD206, IL-10, TGF-β).
- In vitro, KLF15 overexpression enhanced M1-like macrophage polarization, increasing pro-inflammatory mediators (iNOS, CD86, IL-12, TNF-α, IL-1β, IL-6) and decreasing M2 markers (CD206, IL-10, ARG1, TGF-β).
- KLF15 was found to bind the NEK2 promoter in M1-like macrophages, promoting its transcription and activating the NF-κB pathway.
Conclusions:
- KLF15 actively promotes M1 macrophage polarization in DVT, primarily through the NEK2/NF-κB signaling axis.
- KLF15 emerges as a promising therapeutic target for DVT management.
- Further research is necessary to explore the clinical applicability and detailed mechanistic aspects of KLF15 in DVT.
