Effect of purinergic agonists and antagonists on insulin secretion from INS-1 cells (insulinoma cell line) and rat pancreatic islets

Can J Physiol Pharmacol. 2002 Jun;80(6):562-8. doi: 10.1139/y02-079.

Abstract

The effects of purinergic agonists on insulin release are controversial in the literature. In our studies (mainly using INS-1 cells, but also using rat pancreatic islets), ATP had a dual effect on insulin release depending on the ATP concentration: increasing insulin release (EC50 approximately/= 0.0032 microM) and inhibiting insulin release (EC50 approximately/= 0.32 microM) at both 5.6 and 8.3 mM glucose. This is compatible with the view that either two different receptors are involved, or the cells desensitize and (or) the effect of an inhibitory degradation product such as adenosine (ectonucleotidase effect) emerges. The same dual effects of ATP on insulin release were obtained using rat pancreatic islets instead of INS-1 cells. ADPbetaS, which is less degradable than ATP and rather specific for P2Y1 receptors, had a dual effect on insulin release at 8.3 mM glucose: stimulatory (EC50 approximately/= 0.02 microM) and inhibitory (EC50 approximately/= 0.32 microM). The effectiveness of this compound indicates the possible involvement of a P2Y1 receptor. 2-Methylthio-ATP exhibited an insulinotropic effect at very high concentrations (EC50 approximately/= 15 microM at 8.3 mM glucose). This indicated that distinct P2X or the P2Y1 receptor may be involved in these insulin-secreting cells. UTP increased insulin release (EC50 approximately/= 2 microM) very weakly, indicating that a P2U receptor (P2X3 or possibly a P2Y2 or P2Y4) are not likely to be involved. Suramin (50 microM) antagonized the insulinotropic effect of ATP (0.01 microM) and UTP (0.32 microM). Since suramin is not selective, the data indicated that various P2X and P2Y receptors may be involved. PPADS (100 microM), a P2X and P2Y1,4,6 receptor antagonist, was ineffective using either low or high concentrations of ATP and ADPbetaS, which combined with the suramin data hints at a P2Y receptor effect of the compounds. Adenosine inhibited insulin release in a concentration-dependent manner. DPCPX (100 microM), an adenosine (A1) receptor antagonist, inhibited the inhibitory effects of both adenosine and of high concentrations of ATP. Adenosine deaminase (1 U/mL) abolished the inhibitory effect of high ATP concentrations, indicating the involvement of the degradation product adenosine. Repetitive addition of ATP did not desensitize the stimulatory effect of ATP. U-73122 (2 microM), a PLC inhibitor, abolished the ATP effect at low concentrations. The data indicate that ATP at low concentrations is effective via P2Y receptors and the PLC-system and not via P2X receptors; it inhibits insulin release at high concentrations by being metabolized to adenosine.

MeSH terms

  • Adenosine Deaminase / pharmacology
  • Adenosine Diphosphate / analogs & derivatives*
  • Adenosine Diphosphate / pharmacology
  • Adenosine Triphosphate / analogs & derivatives*
  • Adenosine Triphosphate / antagonists & inhibitors
  • Adenosine Triphosphate / pharmacology
  • Animals
  • Cell Line
  • Estrenes / pharmacology
  • Female
  • Glucose / metabolism
  • Insulin / metabolism*
  • Insulinoma / metabolism*
  • Islets of Langerhans / drug effects
  • Islets of Langerhans / metabolism*
  • Male
  • Pancreatic Neoplasms / metabolism*
  • Phosphodiesterase Inhibitors / pharmacology
  • Purinergic Agonists
  • Purinergic Antagonists
  • Purines / agonists*
  • Purines / antagonists & inhibitors*
  • Pyrrolidinones / pharmacology
  • Rats
  • Rats, Wistar
  • Thionucleotides / pharmacology
  • Uridine Triphosphate / pharmacology
  • Xanthines / pharmacology

Substances

  • Estrenes
  • Insulin
  • Phosphodiesterase Inhibitors
  • Purinergic Agonists
  • Purinergic Antagonists
  • Purines
  • Pyrrolidinones
  • Thionucleotides
  • Xanthines
  • 1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione
  • adenosine 5'-O-(2-thiodiphosphate)
  • Adenosine Diphosphate
  • Adenosine Triphosphate
  • 1,3-dipropyl-8-cyclopentylxanthine
  • Adenosine Deaminase
  • Glucose
  • Uridine Triphosphate
  • 2-methylthio-ATP