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Updated: Jun 30, 2026

Homogeneous Time-resolved Förster Resonance Energy Transfer-based Assay for Detection of Insulin Secretion
Published on: May 10, 2018
Dual effect of ATP on glucose-induced insulin secretion in HIT-T15 cells
Dong Hyeon Lee1, Kyu-Sang Park, Dae-Ran Kim
1Department of Physiology, College of Medicine, Pochon CHA University, Seongnam, Gyeonggi-Do, Republic of Korea.
Objectives:
: The study investigated the dual effect of purinergic nucleotides on the secretion of insulin from pancreatic beta cells.
Methods:
: The level of insulin secretion in HIT-T15 cells of static incubation was measured using a radioimmunoassay.
Results:
: The adenine nucleotides reduced the level of glucose-induced insulin secretion in a concentration-dependent manner, and the relative potency order (IC50; muM) was BzATP (6.9) > ATP (20.4) >/= alpha, beta-methylene ATP (23.3) >/= 2-methylthio-ATP (24.9). Suramin and PPADS (200 muM), which are blockers of the purinergic receptors, had a little influence on the activity of ATP. However, the inhibitory effect of ATP was reversed by preincubation with oxidized ATP (200 muM), which is a P2X7 antagonist. The level of insulin secretion in these preincubated cells exposed to the purinergic nucleotides increased in the following order: ATP > alpha, beta-methylene ATP >/= 2-methylthio-ATP. A pretreatment with foskolin and PDBu (100 nM) potentiated the increasing effect of ATP on insulin secretion. The Western blotting showed the expression of P2X7 and P2Y11 receptors.
Conclusions:
: Purinergic stimulation has inhibitory activity on glucose-dependent insulin secretion through the activation of the P2X7 receptor, whereas it has enhancing effect through the activation of the P2Y11 receptor in HIT-T15 cells.
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