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Cardiovascular Research 1997 35(1):168-174; doi:10.1016/S0008-6363(97)00068-0
© 1997 by European Society of Cardiology
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Copyright © 1997, European Society of Cardiology

Adenosine stimulates nitric oxide synthesis in vascular smooth muscle cells

Uichi Ikeda*, Kenji Kurosaki, Ken-ichi Ohya and Kazuyuki Shimada

Department of Cardiology, Jichi Medical School, Minamikawachi-Machi, Tochigi 329-04, Japan

* Corresponding author. Tel.: +81 (285) 44-2111, ext. 3556; fax: +81 (285) 44-5317; e-mail: uikeda@jichi.ac.jp

Objective: The aim was to investigate the effects of adenosine on nitric oxide (NO) synthesis in vascular smooth muscle cells. Methods: NO and cAMP synthesis was measured in confluent rat vascular smooth muscle cells in culture at passage 5–10, using Griess reagent and an enzyme immunoassay kit, respectively. The expression of inducible NO synthase mRNA was assayed by Northern blotting. Results: Incubation of cultures with interleukin-1β (10 ng/ml) for 24 h caused a significant increase in nitrite production. The interleukin-1β-induced nitrite production by vascular smooth muscle cells was significantly increased by adenosine or its stable analogue, 2-chloroadenosine, in a dose-dependent manner. The adenosine A2a receptor antagonist, KF17837, but not the A1 receptor antagonist, DPCPX, significantly inhibited 2-chloroadenosine-mediated nitrite production. The 2-chloroadenosine-induced nitrite production by interleukin-1β-stimulated cells was accompanied by increased inducible NO synthase mRNA accumulation. In the presence of dibutyryl-cAMP (1 mM), interleukin-1β-induced nitrite accumulation was further increased, but the effect of 2-chloroadenosine was not additive or synergistic. Addition of 2-chloroadenosine dose-dependently increased intracellular cAMP levels of vascular smooth muscle cells. Conclusions: These results indicate that adenosine acts on A2 receptors and augments NO synthesis in interleukin-1β-stimulated vascular smooth muscle cells, at least partially through a cAMP-dependent pathway.

KEYWORDS Rat; Adenosine; Interleukin-1; Nitric oxide; cAMP; Vascular smooth muscle cell


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