Pipeleers D G, Schuit F C, in't Veld P A, Maes E, Hooghe-Peters E L, Van de Winkel M, Gepts W
Endocrinology. 1985 Sep;117(3):824-33. doi: 10.1210/endo-117-3-824.
Single pancreatic B cells are purified by autofluorescence-activated cell sorting, and their secretory activity is measured after overnight culture. Compared to intact islets, the isolated cells release 2-fold more insulin under basal conditions and 5-fold less during nutrient stimulation. Their secretory activity can be induced by glucose, leucine, or arginine, but only 0.3-1.7% of their hormone content is liberated at 20 mM nutrient concentrations. This poor nutrient-induced insulin release from purified B cells is attributed to their low cAMP levels and is markedly increased after addition of (Bu)2cAMP, of glucagon, or of pancreatic A cells. These results strongly support the concept that the potent in vivo insulin-releasing action of glucose and leucine is not only dependent on their fuel capacity in pancreatic B cells but also on the concurrent cAMP levels in these cells. In isolated islets, endogenously released glucagon apparently determines the cAMP production in B cells and thus participates in the nutrient-induced secretory process. Somatostatin and epinephrine were shown to exert their suppressive effects via the glucagon-dependent messenger system. It is concluded that nutrients and hormones interact with two different messenger systems which amplify each others' stimulatory effect upon insulin release. cAMP might represent the hormone-induced messenger which sets the B cell's sensitivity and secretory capacity for nutrient stimuli such as glucose. The higher insulin secretory response observed after reaggregation of single B cells could not be attributed to an altered activity in the nutrient or hormonal regulatory units, raising the possibility that the aggregated state of the cells is rather responsible for a better organization or cooperation of the secretory effector unit.
通过自发荧光激活细胞分选法纯化单个胰腺β细胞,并在过夜培养后测量其分泌活性。与完整胰岛相比,分离出的细胞在基础条件下释放的胰岛素多2倍,而在营养刺激下则少5倍。它们的分泌活性可由葡萄糖、亮氨酸或精氨酸诱导,但在20 mM营养物浓度下,仅0.3 - 1.7%的激素含量被释放。纯化的β细胞对营养物诱导的胰岛素释放较差归因于其低cAMP水平,在添加(Bu)2cAMP、胰高血糖素或胰腺α细胞后显著增加。这些结果有力地支持了这样的概念,即葡萄糖和亮氨酸在体内强大的胰岛素释放作用不仅取决于它们在胰腺β细胞中的供能能力,还取决于这些细胞中同时存在的cAMP水平。在分离的胰岛中,内源性释放的胰高血糖素显然决定了β细胞中cAMP的产生,从而参与了营养物诱导的分泌过程。已表明生长抑素和肾上腺素通过依赖胰高血糖素的信使系统发挥其抑制作用。结论是营养物和激素与两种不同的信使系统相互作用,这两种系统相互放大对胰岛素释放的刺激作用。cAMP可能代表激素诱导的信使,它设定了β细胞对葡萄糖等营养刺激的敏感性和分泌能力。单个β细胞重新聚集后观察到的较高胰岛素分泌反应不能归因于营养或激素调节单位活性的改变,这增加了细胞聚集状态更负责分泌效应单位更好的组织或协作的可能性。