Abstract
The purpose of this investigation was to test the hypothesis that A1 receptors modulate extracellular levels of adenosine in cardiovascular tissues. Rat cardiac fibroblasts and human aortic vascular smooth muscle cells were cultured to confluence and various pharmacological agents were applied to the cultures. The extracellular fluid was extracted and adenosine concentrations were measured by HPLC. Three selective A1 receptor antagonists, namely 8-cyclopentyl-1,3-dipropylxanthine, xanthine amine congener, and N-0840, at a concentration of 10 nM significantly increased extracellular levels of adenosine in both rat cardiac fibroblasts and human aortic vascular smooth muscle cells. Further studies in rat cardiac fibroblasts revealed that the effects of A1receptor blockade on extracellular adenosine levels were concentration dependent and prevented by inhibition of Gi proteins with pertussis toxin or blockade of ecto-5′-nucleotidase with α,β-methyleneadenosine-5′-diphosphate. In cardiac fibroblasts in which the extracellular levels of endogenous adenosine were increased, the ability of A1 receptor blockade to augment extracellular adenosine was attenuated. A time-course study revealed a time lag of several hours between blockade of A1 receptors and increases in extracellular adenosine levels. These data suggest that A1 receptors function to detect the long-term levels of extracellular adenosine, and appropriately adjust extracellular adenosine levels by a slow-onset mechanism involving Giproteins and ecto-5′nucleotidase.
Footnotes
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Send reprint requests to: Dr. Edwin K. Jackson, Center for Clinical Pharmacology, University of Pittsburgh Medical Center, 623 Scaife Hall, 200 Lothrop St., Pittsburgh, PA 15213-2582. E-mail:edj+{at}pitt.edu
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↵1 This work was supported by National Institutes of Health Grants HL55314 and HL35909.
- Abbreviations:
- AMPCP
- α,β-methyleneadenosine-5′-diphosphate
- DPCPX
- 8-cyclopentyl-1,3-dipropylxanthine
- XAC
- xanthine amine congener
- Received March 25, 1999.
- Accepted June 1, 1999.
- The American Society for Pharmacology and Experimental Therapeutics
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