db/db小鼠胰岛的分离与特征分析  

Isolation and characterization of pancreatic islets in db/db mice

作　　者：叶泽岚 吴莹 宋益康 林悦 李蓝齐 蔡娟娟[3] 杨超 傅红兴 YE Zelan;WU Ying;SONG Yikang;LIN Yue;LI Lanqi;CAI Juanjuan;YANG Chao;FU Hongxing(Shulan International Medical College,Zhejiang Shuren University,Hangzhou 310015,China;Department of Endocrinology,Sir Run Run Shaw Hospital,Zhejiang University School of Medicine,Hangzhou 310020,China;Department of Pharmacy,the Affiliated Hospital of Medical School,Ningbo University,Ningbo,Zhejiang 315020,China)

机构地区：[1]浙江树人学院树兰国际医学院,浙江杭州312028 [2]浙江大学医学院附属邵逸夫医院内分泌科,浙江杭州310020 [3]宁波大学医学院附属医院药学部,浙江宁波315020

出　　处：《肝胆胰外科杂志》2023年第4期230-232,251,共4页Journal of Hepatopancreatobiliary Surgery

摘　　要：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。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 inβcells 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.

关 键 词：DB/DB小鼠 胰岛 分离 鉴定 

分 类 号：R335[医药卫生—人体生理学] R587[医药卫生—基础医学]