Related Experiment Video
Updated: Sep 24, 2025
A Macrophage Reporter Cell Assay to Examine Toll-Like Receptor-Mediated NF-kB/AP-1 Signaling on Adsorbed Protein Layers on Polymeric Surfaces
Published on: January 7, 2020
Prototypic P2X7 Receptor Agonist, BzATP, Induced the Expression of Unfolded Protein Response Genes in Human M1
Maryam Akhtari1, Seyed Jalal Zargar2, Ali Javinani3
1Department of Cell and Molecular Biology, School of Biology, College of Science, University of Tehran, Tehran, Iran AND Rheumatology Research Center, Tehran University of Medical Sciences, Tehran, Iran AND Inflammation Research Center, Tehran University of Medical Sciences, Tehran, Iran. m.akhtari@ut.ac.ir.
Abstract:
Purinergic receptors stimulation by adenosine triphosphate (ATP) contributes significantly to macrophage activation, and also macrophage cell death. Upon the macrophage activation, the protein load of the endoplasmic reticulum is increased which is resulted in the activation of unfolded protein response (UPR). In the current study, we aimed to evaluate the connection between prototypic P2X7 receptor agonist, extracellular 2'(3')-O-(4-Benzoylbenzoyl)-ATP (BzATP), and the UPR pathway in macrophages. The monocyte-derived macrophages from blood samples of 14 healthy volunteers were skewed toward M1 macrophages after incubation with LPS and IFN-γ. M1 macrophages were treated with 200 µM BzATP. The expression levels of UPR genes, including CHOP, HERP, GADD34, XBP1, and ATF6 in macrophages before and after treatment were measured using real-time polymerase chain reaction. The results demonstrated that the expression of CHOP, HERP, and ATF6 is significantly decreased and the expression level of GADD34 and XBP1 is significantly increased after M1 polarization. BzATP not only significantly increased the expression levels of CHOP, GADD34, ATF6, and HERP but also significantly decreases the XBP1 expression level in M1 macrophages. The present study showed that BzATP induces cellular stress in M1 macrophages by elevating the expression levels of UPR genes including CHOP, GADD34, ATF6, and reducing cell viability.
Insights
Extracellular ATP (BzATP) activates purinergic receptors, triggering unfolded protein response (UPR) gene expression and cellular stress in M1 macrophages. This study reveals BzATP
Area of Science:
- Immunology
- Cell Biology
- Molecular Biology
Background:
- Purinergic receptors, stimulated by adenosine triphosphate (ATP), are crucial for macrophage activation and cell death.
- Macrophage activation leads to increased endoplasmic reticulum protein load, initiating the unfolded protein response (UPR).
Purpose of the Study:
- To investigate the relationship between the P2X7 receptor agonist, 2'(3')-O-(4-Benzoylbenzoyl)-ATP (BzATP), and the UPR pathway in M1 macrophages.
Main Methods:
- Monocyte-derived macrophages were differentiated into M1 macrophages.
- M1 macrophages were treated with 200 µM BzATP.
- Real-time polymerase chain reaction was used to measure UPR gene expression (CHOP, HERP, GADD34, XBP1, ATF6).
Main Results:
- M1 polarization altered UPR gene expression, decreasing CHOP, HERP, and ATF6 while increasing GADD34 and XBP1.
- BzATP treatment significantly increased CHOP, GADD34, ATF6, and HERP expression.
- BzATP treatment significantly decreased XBP1 expression in M1 macrophages.
Conclusions:
- BzATP induces cellular stress in M1 macrophages by modulating UPR gene expression.
- BzATP elevates the expression of CHOP, GADD34, and ATF6, while decreasing XBP1.
- BzATP treatment leads to reduced cell viability in M1 macrophages.
Related Concept Videos
01:37The Unfolded Protein Response
01:31Regulation of the Unfolded Protein Response