cDNA microarray analysis reveals a nuclear factor-kappaB-independent regulation of macrophage function by adenosine.
Adenosine is released into the extracellular space from nerve terminals and cells subjected to ischemic stress. This nucleoside modulates a plethora of cellular functions via occupancy of specific receptors. Adenosine is also an important endogenous regulator of macrophage function, because it suppresses the production of a number of proinflammatory cytokines by these cells. However, the mechanisms of this anti-inflammatory effect have not been well characterized. We hypothesized that adenosine may exert some of its anti-inflammatory effects by decreasing activation of the transcription factor nuclear factor-kappaB (NF-kappaB), because gene expression of most of the proinflammatory cytokines inhibited by adenosine is dependent on NF-kappaB activation. Using bacterial lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages, we found that adenosine as well as adenosine receptor agonists decreased the production of tumor necrosis factor (TNF)-alpha, a typical NF-kappaB-regulated cytokine. This effect of adenosine was not due to an action on the process of TNF-alpha release, because adenosine suppressed also the intracellular levels of TNF-alpha. However, cDNA microarray analysis revealed that mRNA levels of neither TNF-alpha nor other cytokines were altered by adenosine in either LPS-activated or quiescent macrophages. In addition, although LPS induced expression of a number of other, noncytokine genes, including the adenosine A2b receptor, adenosine did not affect the expression of these genes. Furthermore, adenosine as well as adenosine receptor agonists failed to decrease LPS-induced NF-kappaB DNA binding, NF-kappaB promoter activity, p65 nuclear translocation, and inhibitory kappaB degradation. Together, our results suggest that the anti-inflammatory effects of adenosine are independent of NF-kappaB.
Németh
Zoltán H
ZH
Department of Surgery, University of Medicine and Dentistry-New Jersey Medical School, Newark, NJ 07103, USA.
Leibovich
S Joseph
SJ
Deitch
Edwin A
EA
Vizi
E Sylvester
ES
Szabó
Csaba
C
Hasko
Gyorgy
G
D000241:Adenosine / Q000494:pharmacology*; D000818:Animals; D002460:Cell Line; D004347:Drug Interactions; D015870:Gene Expression / Q000187:drug effects; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects* / Q000502:physiology; D051379:Mice; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D043704:Receptor, Adenosine A2B; D018047:Receptors, Purinergic P1 / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
D008070:Lipopolysaccharides; D016328:NF-kappa B; D043704:Receptor, Adenosine A2B; D018047:Receptors, Purinergic P1; D014409:Tumor Necrosis Factor-alpha; D000241:Adenosine