Adenosine and its receptor agonists regulate nitric oxide production and RAW 264.7 macrophages via both receptor binding and its downstream metabolites-inosine.
Adenosine and its receptor agonists enhanced the production of nitric oxide (NO) in lipopolysaccharide (LPS)-treated RAW 264.7 cells. The enhancement of LPS-induced NO production by adenosine, as represented by the amount of its oxidation products, nitrite and nitrate, was inhibited by adenosine uptake inhibitors, such as dipyridamole, S(4-nitrobenzyl)-6-thioinosine (NBTI) and S(4-nitrobenzyl)-6-thioguanosine (NBTG). These indicate that the uptake of adenosine by macrophages is a prerequisite for the enhancement effects observed. A downstream metabolite of adenosine, inosine, also potentiated the LPS-induced NO production in a dose-dependent manner while its enhancement effect was also inhibited by dipyridamole. However, the degree of enhancement by inosine on NO production and nitric oxide synthase (NOS) activity in LPS-treated RAW 264.7 was weaker than the effect of adenosine. Furthermore, adenosine agonists also enhanced the NO production in a dose-dependent manner, but were not specific for A1, A2 nor A3 adenosine receptor. Adenosine uptake inhibitors had no effects on the enhancement activity of the adenosine receptor agonists. Thus, extracellular receptor/s may also play an important role in the observed enhancement responses. The results of this study indicate that the enhancement effects of adenosine on NO production in macrophages could be mediated by the extracellular adenosine receptors as well as the downstream metabolites of adenosine.
Min
H W
HW
Department of Medicine, National University of Singapore and Defence Medical Research Institute, Singapore. mdchonwm@nus.edu.sg
Moochhala
S
S
Eng
K H
KH
D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000241:Adenosine / Q000494:pharmacology / Q000502:physiology*; D001427:Bacterial Toxins / Q000494:pharmacology; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004398:Dyneins / Q000037:antagonists & inhibitors; D004768:Enterotoxins / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins; D006801:Humans; D019271:Hypoxanthine / Q000494:pharmacology; D007288:Inosine / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D058906:Purinergic P1 Receptor Agonists
D001427:Bacterial Toxins; D004768:Enterotoxins; D004791:Enzyme Inhibitors; D029968:Escherichia coli Proteins; D008070:Lipopolysaccharides; D058906:Purinergic P1 Receptor Agonists; D019271:Hypoxanthine; D009569:Nitric Oxide; C010192:9-(2-hydroxy-3-nonyl)adenine; D007288:Inosine; C027538:heat-labile enterotoxin, E coli; D019001:Nitric Oxide Synthase; D004398:Dyneins; D000225:Adenine; D000241:Adenosine