Intracellular signaling pathway inhibitors suppress inflammatory response induced by synergistically acting bacterial substances
AbstractIntroduction. Combined activation of NOD-like and Toll-like receptors of innate immune cells is a potent pro-inflammatory stimulus that may underlie the systemic inflammatory response in bacterial sepsis.
Aim – to evaluate the possibility to suppress pro-inflammatory cytokine production by macrophages activated with NOD1 and TLR4 agonist combination using inhibitors of NF-κB-dependent and mitogen-activated protein kinase (MAPK)-dependent signaling pathways.
Material and methods. Macrophages generated from healthy donor monocytes were stimulated with NOD1 and TLR4 agonists separately or simultaneously. The activity of NF-κB-dependent and MAPK-dependent signaling pathways was suppressed using low-molecular-weight kinase inhibitors that were added before or after commencement of cell activation. As read-outs, we measured mRNA expression of pro-inflammatory cytokines TNF, IL6, IL12B and IL23A using RT-PCR as well as secretion of TNF, IL-6 and IL-23 using ELISA.
Results. Inhibition of NF-κB-dependent and MAPK-dependent signaling pathways allowed to prevent or interrupt expression and production of TNF, IL-6 and IL-23 by macrophages activated by a potent pro-inflammatory stimulus, a NOD1 + TLR4 agonist combination. Most effective was simultaneous inhibition of both signaling pathways. We also show contribution of NF-κB-dependent and MAPK-dependent signaling pathways to synergistic enhancement of IL12B gene expression.
Conclusions. These results obtained can be utilized when developing novel approaches to anti-inflammatory therapy of severe bacterial infections.
Keywords:macrophages; NOD1; TLR4; muramyl peptides; lipopolysaccharide; mitogen-activated protein kinases; nuclear factor-κB; inhibitors
For citation: Masyutina A.M., Murugin V.V., Pashenkov M.V. Intracellular signaling pathway inhibitors suppress inflammatory response induced by synergistically acting bacterial substances. Immunologiya. 2024; 45 (1): 21–32. DOI: https://doi.org/10.33029/1816-2134-2024-45-1-21-32 (in Russian)
Funding. This work was supported by the Russian Science Foundation grant No. 21-15-00211.
Conflict of interests. The authors declare no conflict of interests.
Authors’ contribution. Conducting experiments and analysing results – Masyutina A.M., Murugin V.V.; designing experiments, analysing results and writing the article – Pashenkov M.V.
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