Glucose- and time-dependence of islet amyloid formation in vitro.
Glucose- and time-dependence of islet amyloid formation in vitro.
复制标题
体外胰岛淀粉样蛋白形成的葡萄糖和时间依赖性。
DOI：
10.1016/j.bbrc.2006.12.187
复制
发表时间：
2007
影响因子：
3.1
通讯作者：
Kahn,StevenE
中科院分区：
文献类型：
--
作者：
Zraika,Sakeneh;Hull,RebeccaL;Udayasankar,Jayalakshmi;Utzschneider,KristinaM;Tong,Jenny;Gerchman,Fernando;Kahn,StevenE
胰岛淀粉样蛋白会导致 2 型糖尿病中 β 细胞质量的损失。为了研究葡萄糖和时间对淀粉样蛋白形成的作用，我们使用从人胰岛淀粉样蛋白多肽（hIAPP）转基因小鼠中分离的胰岛开发了一种快速体外模型。来自 hIAPP 转基因和非转基因小鼠的胰岛在 5.5、11.1、16.7 或 33.3mmol/l 葡萄糖的条件下培养长达 7 天。在整个培养期间的不同时间点，收获胰岛以测定淀粉样蛋白和β细胞面积，并测量细胞活力、胰岛素含量和分泌。 hIAPP转基因胰岛在16.7或33.3mmol/l葡萄糖中培养后，与5.5或11.1mmol/l葡萄糖培养物相比，淀粉样蛋白形成显着增加。淀粉样蛋白早在第 2 天就被检测到，并以时间依赖性方式增加，因此到第 7 天，hIAPP 转基因胰岛中 β 细胞面积的比例明显下降。在16.7mmol/l葡萄糖中培养7天后，与非转基因胰岛相比，hIAPP转基因胰岛的活力降低了24%，β细胞面积和胰岛素含量减少，但胰岛素分泌没有变化。因此，我们开发了一种光学显微镜可见的胰岛淀粉样蛋白形成的快速体外模型，该模型既依赖于葡萄糖又依赖于时间。该模型中淀粉样蛋白的形成与细胞活力降低和β细胞损失有关，但与足够的功能适应有关。因此，它使得研究 2 型糖尿病中与淀粉样蛋白相关的 β 细胞质量损失的机制成为可能。
Islet amyloid contributes to the loss of β-cell mass in type 2 diabetes. To examine the roles of glucose and time on amyloid formation, we developed a rapid in vitro model using isolated islets from human islet amyloid polypeptide (hIAPP) transgenic mice. Islets from hIAPP transgenic and non-transgenic mice were cultured for up to 7 days with either 5.5, 11.1, 16.7 or 33.3mmol/l glucose. At various time-points throughout the culture period, islets were harvested for determination of amyloid and β-cell areas, and for measures of cell viability, insulin content, and secretion. Following culture of hIAPP transgenic islets in 16.7 or 33.3mmol/l glucose, amyloid formation was significantly increased compared to 5.5 or 11.1mmol/l glucose culture. Amyloid was detected as early as day 2 and increased in a time-dependent manner so that by day 7, a decrease in the proportion of β-cell area in hIAPP transgenic islets was evident. When compared to non-transgenic islets after 7-day culture in 16.7mmol/l glucose, hIAPP transgenic islets were 24% less viable, had decreased β-cell area and insulin content, but displayed no change in insulin secretion. Thus, we have developed a rapid in vitro model of light microscopy-visible islet amyloid formation that is both glucose- and time-dependent. Formation of amyloid in this model is associated with reduced cell viability and β-cell loss but adequate functional adaptation. It thus enables studies investigating the mechanism(s) underlying the amyloid-associated loss of β-cell mass in type 2 diabetes.