ADP mediates inhibition of insulin secretion by activation of P2Y13 receptors in mice
ADP mediates inhibition of insulin secretion by activation of P2Y13 receptors in mice
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DOI：
10.1007/s00125-010-1807-8
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发表时间：
2010-09-01
期刊：
影响因子：
8.2
通讯作者：
Erlinge, D.
中科院分区：
文献类型：
--
作者：
Amisten, S.;Meidute-Abaraviciene, S.;Erlinge, D.
目的/假设研究细胞外嘌呤对小鼠胰岛分泌胰岛素的影响方法分离小鼠胰岛和β细胞，采用mRNA实时定量、cAMP定量、胰岛素和胰高血糖素分泌等方法进行检测。在MIN 6c 4细胞中测量ATP释放。胰岛素和胰高血糖素的分泌测定在体内葡萄糖injection.Results酶去除细胞外ATP在低葡萄糖水平增加胰岛素和胰高血糖素的分泌，而在高葡萄糖水平胰岛素分泌减少，胰高血糖素分泌刺激，表明嘌呤的自分泌效应。在MIN 6c 4细胞中，显示葡萄糖确实诱导ATP释放到细胞外空间中。实时定量PCR证实了ADP受体P2 Y（1）和P2 Y（13）在完整小鼠胰岛和分离的β细胞中的表达。稳定的ADP类似物2-MeSADP对胰岛素分泌无影响。然而，与P2 Y（1）拮抗剂MRS 2179共孵育抑制胰岛素分泌，而与P2 Y（13）拮抗剂MRS 2211共孵育刺激胰岛素分泌，表明ADP通过P2 Y（1）作用刺激胰岛素分泌，而通过P2 Y（13）信号传导抑制胰岛素分泌。通过MRS 2211本身的P2 Y13拮抗作用增加了中等（8.3 mmol/l）和高（20 mmol/l）葡萄糖水平下胰岛素和胰高血糖素的分泌，证实了ADP的自分泌作用。在体内葡萄糖注射期间施用MRS 2211导致胰岛素分泌增加和葡萄糖水平降低。结论/解释总之，ADP作用于P2 Y（13）受体抑制胰岛素释放。P2 Y拮抗剂（13）增加胰岛素释放，可用于治疗糖尿病。
Aims/hypotheses To investigate the effects of extracellular purines on insulin secretion from mouse pancreatic islets.Methods Mouse islets and beta cells were isolated and examined with mRNA real-time quantification, cAMP quantification and insulin and glucagon secretion. ATP release was measured in MIN6c4 cells. Insulin and glucagon secretion were measured in vivo after glucose injection.Results Enzymatic removal of extracellular ATP at low glucose levels increased the secretion of both insulin and glucagon, while at high glucose levels insulin secretion was reduced and glucagon secretion was stimulated, indicating an autocrine effect of purines. In MIN6c4 cells it was shown that glucose does induce release of ATP into the extracellular space. Quantitative real-time PCR demonstrated the expression of the ADP receptors P2Y(1) and P2Y(13) in both intact mouse pancreatic islets and isolated beta cells. The stable ADP analogue 2-MeSADP had no effect on insulin secretion. However, co-incubation with the P2Y(1) antagonist MRS2179 inhibited insulin secretion, while co-incubation with the P2Y(13) antagonist MRS2211 stimulated insulin secretion, indicating that ADP acting via P2Y(1) stimulates insulin secretion, while signalling via P2Y(13) inhibits the secretion of insulin. P2Y13 antagonism through MRS2211 per se increased the secretion of both insulin and glucagon at intermediate (8.3 mmol/l) and high (20 mmol/l) glucose levels, confirming an autocrine role for ADP. Administration of MRS2211 during glucose injection in vivo resulted in both increased secretion of insulin and reduced glucose levels.Conclusions/interpretation In conclusion, ADP acting on the P2Y(13) receptors inhibits insulin release. An antagonist to P2Y(13) increases insulin release and could be evaluated for the treatment of diabetes.