Department of Biochemistry and Biophysics and Institute for Diabetes, Obesity, and Metabolism, University of Pennsylvania School of Medicine, Philadelphia, 19104-6140, USA.
Am J Physiol Endocrinol Metab. 2010 Sep;299(3):E475-85. doi: 10.1152/ajpendo.00072.2010. Epub 2010 Jul 6.
Fatty acids, acetylcholine, and GLP-1 enhance insulin secretion in a glucose-dependent manner. However, the interplay between glucose, fatty acids, and the neuroendocrine regulators of insulin secretion is not well understood. Therefore, we studied the acute effects of PA (alone or in combination with glucose, acetylcholine, or GLP-1) on isolated cultured mouse islets. Two different sets of experiments were designed. In one, a fixed concentration of 0.5 mM of PA bound to 0.15 mM BSA was used; in the other, a PA ramp from 0 to 0.5 mM was applied at a fixed albumin concentration of 0.15 mM so that the molar PA/BSA ratio changed within the physiological range. At a fixed concentration of 0.5 mM, PA markedly inhibited acetylcholine-stimulated insulin release, the rise of intracellular Ca(2+), and enhancement of cAMP production but did not influence the effects of GLP-1 on these parameters of islet cell function. 2-ADB, an IP(3) receptor inhibitor, reduced the effect of acetylcholine on insulin secretion and reversed the effect of PA on acetylcholine-stimulated insulin release. Islet perfusion for 35-40 min with 0.5 mM PA significantly reduced the calcium storage capacity of ER measured by the thapsigargin-induced Ca(2+) release. Oxygen consumption due to low but not high glucose was reduced by PA. When a PA ramp from 0 to 0.5 mM was applied in the presence of 8 mM glucose, PA at concentrations as low as 50 microM significantly augmented glucose-stimulated insulin release and markedly reduced acetylcholine's effects on hormone secretion. We thus demonstrate that PA acutely reduces the total oxygen consumption response to glucose, glucose-dependent acetylcholine stimulation of insulin release, Ca(2+), and cAMP metabolism, whereas GLP-1's actions on these parameters remain unaffected or potentiated. We speculate that acute emptying of the ER calcium by PA results in decreased glucose stimulation of respiration and acetylcholine potentiation of insulin secretion.
脂肪酸、乙酰胆碱和 GLP-1 以葡萄糖依赖的方式增强胰岛素分泌。然而,葡萄糖、脂肪酸和胰岛素分泌的神经内分泌调节剂之间的相互作用还不是很清楚。因此,我们研究了 PA(单独或与葡萄糖、乙酰胆碱或 GLP-1 联合)对分离培养的小鼠胰岛的急性作用。设计了两组不同的实验。一组使用固定浓度为 0.5 mM 的与 0.15 mM BSA 结合的 PA;另一组在固定的白蛋白浓度 0.15 mM 下应用从 0 到 0.5 mM 的 PA 斜坡,使摩尔 PA/BSA 比在生理范围内变化。在固定浓度为 0.5 mM 时,PA 明显抑制乙酰胆碱刺激的胰岛素释放、细胞内 Ca(2+)的增加和 cAMP 产生的增强,但不影响 GLP-1 对胰岛细胞功能这些参数的影响。IP(3)受体抑制剂 2-ADB 降低了乙酰胆碱对胰岛素分泌的作用,并逆转了 PA 对乙酰胆碱刺激的胰岛素释放的作用。用 0.5 mM PA 对胰岛进行 35-40 分钟的灌注,显著降低了内质网钙储存能力,这是通过 thapsigargin 诱导的 Ca(2+)释放来测量的。低但不是高葡萄糖引起的耗氧量因 PA 而减少。当在 8 mM 葡萄糖存在下应用从 0 到 0.5 mM 的 PA 斜坡时,浓度低至 50 microM 的 PA 显著增强了葡萄糖刺激的胰岛素释放,并显著降低了乙酰胆碱对激素分泌的作用。因此,我们证明 PA 急性降低了对葡萄糖的总耗氧量反应、葡萄糖依赖的乙酰胆碱刺激的胰岛素释放、Ca(2+)和 cAMP 代谢,而 GLP-1 对这些参数的作用保持不变或增强。我们推测,PA 急性排空内质网钙导致葡萄糖刺激呼吸和乙酰胆碱增强胰岛素分泌减少。