db/db小鼠胰岛的分离与特征分析

目的 分离db/db小鼠胰岛，并对其特征进行检测和分析。方法 采用胶原酶V对10只db/db小鼠胰岛分离进行逆行胰管灌注和消化，用Ficoll-1077和Ficoll-1119进行胰岛的不连续密度梯度纯化，对分离的胰岛进一步进行手工挑选，并用DTZ进行胰岛和纯度鉴定，用透射电镜观察胰岛内部分泌颗粒等情况。结果 经本方法分离可得到db/db小鼠胰岛数量为（122.4±6.6）个/只，当量为（483.6±82.3）IEQ/只，与ICR小鼠胰岛分离结果差异有统计学意义（P<0.05）;db/db小鼠胰岛DTZ染色后显淡红色；胰岛大小指数为（3.96±0.64），显著大于ICR小鼠胰岛（P<0.05）；透射电镜下显示β细胞中的胰岛素分泌颗粒减少，分泌颗粒颜色较浅。结论 采用本方法可分离得到db/db小鼠胰岛，为后续开展基于胰岛功能和特征变化的2型糖尿病患者治疗研究提供一种参考。

Isolation and characterization of pancreatic islets in db/db mice

Objective To isolate and identify the characteristics of islets in db/db mice. Methods The db/ db mice pancreatic islets were digested with injecting collagenase V into pancreatic ductal, and were purified by discontinuous density gradient purification and Ficoll, followed by further hand-selected. The identification of islets was determined by dithizone (DTZ) staining. The secreted particles inside the islets were observed by transmission electron microscopy (TEM). Results Each db/db mice could yield (122.4±6.6) islets and (483.6±82.3) islet equivalents, which was significantly different from the results of ICR mice (P<0.05). The islets from db/db mice showed reddish after DTZ staining. The islet size index from db/db mice was (3.96±0.64), which was significantly higher than that of ICR mice (P<0.05). The insulin secretory granules in β cells from db/db mice were reduced and lighter in color via the TEM observation. Conclusion Based on the function and characteristics of the islets, the method of yielding db/db mice pancreatic islets, can provide a good foundation for the subsequent study on treatment of type 2 diabetes.