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Dimethyl Pent-2-Enedioate inhibits LPS-induced inflammatory response in macrophages
Zhi-Ying Zhou1, Zhi-Peng Zhou2, Ying-Xing Yue3
1Affiliated Zhejiang Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China; Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China.
Background:
Endogenous metabolite itaconate and its derivative Dimethyl itaconate (DMI) exhibit significant anti-inflammatory effects. Dimethyl Pent-2-Enedioate (DMP), an isomer of DMI, may possess similar properties. This study investigates the anti-inflammatory effects of DMP in LPS-induced macrophages and explores its potential regulatory mechanisms.
Methods:
Inflammatory marker levels were assessed at both the mRNA and protein levels using ELISA and qRT-PCR. The activation status of macrophages was evaluated by flow cytometry, quantifying the number of CD40-positive cells. RNA sequencing was conducted to investigate the transcriptomic changes following DMP treatment. Subsequent GO and KEGG enrichment analyses were performed to identify potential mechanisms underlying DMP's effects. Western blot analysis was employed to assess the expression of p-p65, while immunofluorescence analysis was used to examine p65 nuclear translocation, providing insight into the regulatory effects of DMP on the NF-κB signaling pathway.
Results:
DMP inhibited the expression of inflammatory markers TNF-α, IL-6, and MCP-1 at both mRNA and protein levels. Flow cytometry analysis revealed a decrease in CD40-positive cells. RNA sequencing identified DEGs enriched in inflammation-related pathways. Western blotting and immunofluorescence confirmed that DMP reduced p-p65 expression and inhibited p65 nuclear translocation, suggesting a potential regulatory effect on the NF-κB signaling pathway.
Conclusion:
DMP significantly inhibits LPS-induced inflammation in macrophages, with its underlying mechanisms being complex. Our data demonstrate that DMP exerts its anti-inflammatory effects at least in part through the downregulation of the NF-κB pathway, offering potential applications in the prevention and treatment of inflammation-related diseases.
Insights
Dimethyl Pent-2-Enedioate (DMP) effectively reduces inflammation in macrophages by inhibiting key inflammatory markers. This study reveals DMP’s anti-inflammatory action is partly mediated by downregulating the NF-κB pathway.
Area of Science:
- Immunology
- Molecular Biology
- Pharmacology
Background:
- Itaconate and Dimethyl itaconate (DMI) are known for anti-inflammatory properties.
- Dimethyl Pent-2-Enedioate (DMP), a DMI isomer, is investigated for similar effects.
- LPS-induced macrophages serve as a model to study DMP's anti-inflammatory potential.
Purpose of the Study:
- To investigate the anti-inflammatory effects of DMP in LPS-induced macrophages.
- To explore the molecular mechanisms underlying DMP's anti-inflammatory actions.
- To assess DMP's impact on inflammatory markers and signaling pathways.
Main Methods:
- Assessed inflammatory markers (TNF-α, IL-6, MCP-1) via ELISA and qRT-PCR.
- Evaluated macrophage activation using flow cytometry (CD40 marker).
- Utilized RNA sequencing, GO/KEGG analysis, Western blot (p-p65), and immunofluorescence (p65 translocation) to elucidate mechanisms, focusing on the NF-κB pathway.
Main Results:
- DMP significantly inhibited TNF-α, IL-6, and MCP-1 expression at mRNA and protein levels.
- Reduced CD40-positive cells and identified inflammation-related differentially expressed genes (DEGs).
- Confirmed DMP's downregulation of p-p65 expression and p65 nuclear translocation, indicating NF-κB pathway inhibition.
Conclusions:
- DMP demonstrates significant anti-inflammatory effects in LPS-induced macrophages.
- The anti-inflammatory mechanism involves, at least partly, the downregulation of the NF-κB pathway.
- DMP shows promise for treating inflammation-related diseases.
