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Updated: Aug 12, 2026
Investigation of Macrophage Polarization Using Bone Marrow Derived Macrophages
Published on: June 23, 2013
Pseudane V alleviates ox-LDL-induced macrophage M1 polarization by inhibiting m6A modification of PPARGC1A
1Department of Cardiology, Lanzhou Petrochemical General Hospital (The Fourth Affiliated Hospital of Gansu University of Chinese Medicine), Lanzhou, 730060, China.
Abstract:
Atherosclerosis is driven by oxidized low-density lipoprotein (ox-LDL)-triggered macrophage malfunction, yet the precise pathways and effective counter-measures remain elusive. Here we delineated how N6-methyladenosine (m6A) RNA methylation governs ox-LDL-induced M1 macrophage polarization and evaluated marine natural products for therapeutic intervention. Human monocytic THP-1 cells differentiated into M0 macrophages were treated with ox-LDL. M1/M2 polarization states were analyzed using flow cytometry, and changes in polarization markers were examined using quantitative real-time polymerase chain reaction (qRT-PCR). Global m6A changes were detected using m6A dot blot and quantification analysis. Methylated RNA immunoprecipitation sequencing (MeRIP-seq) and RNA sequencing (RNA-seq) were performed to identify downstream target genes of the ox-LDL-m6A pathway. The role of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PPARGC1A) in ox-LDL-induced M1 polarization was analyzed. A high-throughput marine natural product library was employed to discover agents capable of reversing ox-LDL-driven macrophage polarization. ox-LDL induces macrophage M1 polarization, and M1 macrophages damage endothelial cells. ox-LDL-induced polarization of macrophages into the M1 phenotype was associated with increased global RNA m6A modification. High levels of m6A modification induced by ox-LDL suppressed PPARGC1A expression in a YTH domain family 2 (YTHDF2)-dependent manner, leading to M1 polarization. Pseudane V, a marine natural product, effectively reduced ox-LDL-induced M1 polarization and protected vascular endothelial cells by correcting abnormal m6A modification of PPARGC1A. In conclusion, ox-LDL induces m6A modification of PPARGC1A, suppressing its expression and promoting M1 polarization of macrophages, contributing to atherosclerosis development. Pseudane V counteracts excessive m6A modification caused by ox-LDL, preventing M1 polarization of macrophages. These findings suggest the potential use of Pseudane V in preventing atherosclerosis.
Insights
Oxidized LDL triggers N6-methyladenosine (m6A) RNA changes, promoting M1 macrophage polarization and atherosclerosis. Marine natural product Pseudane V reverses these m6A modifications, offering potential atherosclerosis prevention.
Area of Science:
- Molecular Biology
- Immunology
- Cardiovascular Research
Background:
- Atherosclerosis pathogenesis involves macrophage dysfunction driven by oxidized low-density lipoprotein (ox-LDL).
- The precise molecular mechanisms, including RNA methylation, governing ox-LDL-induced macrophage polarization are not fully understood.
- Targeting macrophage polarization presents a therapeutic strategy for atherosclerosis.
Purpose of the Study:
- To elucidate the role of N6-methyladenosine (m6A) RNA methylation in ox-LDL-induced M1 macrophage polarization.
- To identify downstream targets of the ox-LDL-m6A pathway in macrophages.
- To discover marine natural products capable of reversing ox-LDL-driven macrophage polarization for atherosclerosis intervention.
Main Methods:
- Human monocytic THP-1 cells were differentiated and treated with ox-LDL.
- Macrophage polarization (M1/M2) was assessed via flow cytometry and qRT-PCR.
- Global m6A levels were analyzed using dot blot; MeRIP-seq and RNA-seq identified target genes.
- High-throughput screening of marine natural products was performed.
Main Results:
- Ox-LDL induced M1 macrophage polarization, associated with increased global RNA m6A modification.
- Ox-LDL-induced m6A suppressed PPARGC1A expression in a YTHDF2-dependent manner, promoting M1 polarization.
- Marine natural product Pseudane V reversed ox-LDL-induced M1 polarization and protected endothelial cells by normalizing PPARGC1A m6A modification.
Conclusions:
- Ox-LDL promotes atherosclerosis by increasing m6A modification of PPARGC1A, suppressing its expression and driving M1 macrophage polarization.
- Pseudane V effectively counteracts ox-LDL-induced m6A changes, inhibiting M1 polarization.
- Pseudane V demonstrates potential as a therapeutic agent for preventing atherosclerosis.
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