Detimary P, Xiao C, Henquin J C
Unit of Endocrinology and Metabolism, University of Louvain Faculty of Medicine, UCL 55.30, Avenue Hippocrate 55, B-1200 Brussels, Belgium.
Biochem J. 1997 Jun 1;324 ( Pt 2)(Pt 2):467-71. doi: 10.1042/bj3240467.
Glucose metabolism in pancreatic B-cells leads to an increase in the ATP/ADP ratio that might participate in the regulation of insulin secretion. Good correlations have also been observed between guanine nucleotide levels in isolated pancreatic islets and insulin secretion. To assess whether guanine nucleotides have a specific role in stimulus-secretion coupling, their concentration should be modified selectively. This was attempted by culturing mouse islets overnight in the presence of mycophenolic acid (MPA), an inhibitor of GMP synthesis at the level of IMP dehydrogenase. The drug (25-50 microg/ml) did not affect the insulin content but decreased the GTP content of the islets and inhibited insulin secretion during subsequent incubation in the presence of 15 mM glucose. However, MPA also decreased the ATP/ADP ratio in the islets. The addition of guanine to the culture medium (to stimulate the salvage pathway of GTP synthesis) restored normal GTP levels, corrected the ATP/ADP ratio and partly prevented the inhibition of insulin release. In contrast, attempts to stimulate ATP synthesis specifically (by provision of adenine or adenosine) failed to reverse any of the effects of MPA. It is concluded that guanine and adenine nucleotide pools are tightly linked and cannot be specifically affected by MPA in pancreatic islet cells, probably because of the activity of nucleoside diphosphate kinase and because of the role of GTP in several reactions leading to adenine nucleotide generation. Contrary to previous claims, MPA is not an adequate tool for evaluating a specific role of guanine nucleotides in the control of insulin secretion.
胰腺β细胞中的葡萄糖代谢会导致ATP/ADP比值升高,这可能参与胰岛素分泌的调节。在分离的胰岛中,鸟嘌呤核苷酸水平与胰岛素分泌之间也存在良好的相关性。为了评估鸟嘌呤核苷酸在刺激-分泌偶联中是否具有特定作用,应选择性地改变其浓度。为此,将小鼠胰岛在霉酚酸(MPA)存在下培养过夜,MPA是一种在次黄嘌呤核苷酸脱氢酶水平抑制GMP合成的抑制剂。该药物(25 - 50微克/毫升)不影响胰岛素含量,但降低了胰岛的GTP含量,并在随后15毫摩尔葡萄糖存在的孵育过程中抑制胰岛素分泌。然而,MPA也降低了胰岛中的ATP/ADP比值。向培养基中添加鸟嘌呤(以刺激GTP合成的补救途径)可恢复正常的GTP水平,纠正ATP/ADP比值,并部分防止胰岛素释放的抑制。相反,特异性刺激ATP合成的尝试(通过提供腺嘌呤或腺苷)未能逆转MPA的任何作用。得出的结论是,鸟嘌呤和腺嘌呤核苷酸池紧密相连,在胰岛细胞中MPA不能特异性地影响它们,这可能是由于核苷二磷酸激酶的活性以及GTP在导致腺嘌呤核苷酸生成的几个反应中的作用。与先前的说法相反,MPA不是评估鸟嘌呤核苷酸在胰岛素分泌控制中特定作用的合适工具。