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Updated: May 16, 2026
Investigation of Macrophage Polarization Using Bone Marrow Derived Macrophages
Published on: June 23, 2013
Inhibition of RGS1 ameliorates obesity-associated macrophage dysfunction and metaflammation through restoration of
Yunmeng Li1, Xuan Fu2, Baolong Yin2
1Department of Endocrinology and Metabolism, The First Affiliated Hospital of Shihezi University, Shihezi, 832008, China.
Background:
Obesity-associated metabolic inflammation is characterized by the infiltration and pro-inflammatory polarization of adipose tissue macrophages (ATMs). Mitochondrial dysfunction represents a central pathogenic mechanism; however, the upstream molecular mechanisms that link metabolic stress to impaired mitochondrial quality control in macrophages remain inadequately defined. Regulator of G protein signaling 1 (RGS1) is significantly upregulated in ATMs under obese conditions; nevertheless, its functional role and mechanistic relevance have not been fully elucidated.
Methods:
Integrated transcriptomic analyses identified RGS1 as a pivotal gene associated with obesity, and its expression profiles in adipose tissue macrophages were analyzed by integrating single-cell RNA sequencing data from humans and mice. In vivo, immunohistochemistry and immunofluorescence assessed RGS1 localization in adipose tissue from high-fat diet-fed obese mice. In vitro, RAW264.7 cells and bone marrow-derived macrophages (BMDMs) were treated with palmitic acid (PA) or PA plus interleukin-6 (IL-6), combined with lentivirus-mediated RGS1 knockdown. The effects of RGS1 on mitochondrial integrity, mitophagic flux, lipid metabolism, and macrophage polarization were assessed by Western blot, quantitative real-time PCR, transmission electron microscopy, TMRE staining, reactive oxygen species (ROS) production, ATP levels, and oxidative phosphorylation (OXPHOS). Furthermore, bafilomycin A1 (Baf-A1) was used to assess mitophagic flux, and Mdivi-1 was utilized to inhibit mitophagy for mechanistic validation.
Results:
Bioinformatic analyses revealed a significant upregulation of RGS1 in ATMs under obese conditions. In vivo studies demonstrated elevated levels of RGS1 in the epididymal adipose tissue of obese mice, where it co-localized with CD86+ M1 macrophages. In vitro, PA treatment induced RGS1 expression and caused lipid accumulation and M1 macrophage polarization; co-stimulation with PA and interleukin-6 (IL-6) further upregulated RGS1 and exacerbated pro-inflammatory responses. Conversely, RGS1 knockdown markedly attenuated both lipid accumulation and M1 polarization. Furthermore, PA treatment induced severe mitochondrial dysfunction, characterized by cristae disruption, loss of membrane potential, ROS accumulation, ATP depletion, and decreased expression of OXPHOS complexes, and inhibited PINK1/Parkin-mediated mitophagy. In contrast, RGS1 knockdown restored mitochondrial function. Baf-A1 treatment assays confirmed that RGS1 knockdown augmented mitophagic flux, while treatment with Mdivi-1 significantly diminished this protective effect, indicating that these protective effects were likely mediated in a mitophagy-dependent manner.
Conclusion:
RGS1 acts as a critical switch linking metabolic stress to macrophage dysfunction. Targeting RGS1 may represent a promising therapeutic strategy for alleviating obesity-induced inflammation.
Insights
Regulator of G protein signaling 1 (RGS1) links metabolic stress to macrophage dysfunction in obesity. Targeting RGS1 may reduce obesity-induced inflammation by restoring mitochondrial function and macrophage polarization.
Area of Science:
- Immunology
- Metabolic disease research
- Cellular biology
Background:
- Obesity triggers metabolic inflammation via adipose tissue macrophage (ATM) infiltration and pro-inflammatory polarization.
- Mitochondrial dysfunction is key in obesity pathogenesis, but upstream regulators linking metabolic stress to macrophage mitochondrial issues are unclear.
- Regulator of G protein signaling 1 (RGS1) is upregulated in ATMs during obesity, yet its function remains poorly understood.
Purpose of the Study:
- To investigate the functional role of Regulator of G protein signaling 1 (RGS1) in obesity-associated metabolic inflammation.
- To elucidate the molecular mechanisms by which RGS1 influences macrophage mitochondrial function and polarization under metabolic stress.
Main Methods:
- Integrated transcriptomic analysis to identify RGS1's role in obesity.
- In vivo studies using high-fat diet-fed mice and in vitro experiments with macrophages treated with palmitic acid (PA) and IL-6.
- Assessment of RGS1's impact on mitochondrial integrity, mitophagy, lipid metabolism, and macrophage polarization using molecular and cellular assays.
- Genetic manipulation (RGS1 knockdown) and pharmacological inhibitors (Baf-A1, Mdivi-1) to validate mechanisms.
Main Results:
- RGS1 is significantly upregulated in ATMs of obese mice and promotes M1 macrophage polarization and lipid accumulation.
- Metabolic stress (PA) induces RGS1, leading to mitochondrial dysfunction (impaired cristae, reduced membrane potential, ROS, ATP depletion) and suppressed mitophagy.
- RGS1 knockdown ameliorates lipid accumulation, M1 polarization, and mitochondrial dysfunction, restoring mitophagic flux in a mitophagy-dependent manner.
Conclusions:
- RGS1 acts as a critical mediator linking metabolic stress to macrophage dysfunction in obesity.
- Targeting RGS1 presents a potential therapeutic strategy for mitigating obesity-induced inflammation.
