Exendin-4对不同热缺血时间心死亡供体大鼠胰岛功能的影响  

Exendin-4 effect on pancreatic islet function of donor rats with cardiac death at different heat ischemia time

作　　者：车彦鹏[1] 刘永锋[2] 宋文卿[1] 梁世博[1] 

机构地区：[1]大庆龙南医院普外科,黑龙江省大庆市163453 [2]中国医科大学附属第一医院普通外科教研室肝胆外科,辽宁省沈阳市110001

出　　处：《中国组织工程研究》2014年第36期5747-5751,共5页Chinese Journal of Tissue Engineering Research

摘　　要：背景:心死亡供体大鼠供胰的利用不可避免地经过热缺血损伤,因此热缺血时间的长短对获得的胰岛数量及功能有重要影响。目的:观察Exendin-4对不同热缺血时间心死亡供体大鼠胰岛功能的影响。方法:大鼠胰岛细胞体外培养,根据实验条件随机分为3大组:热缺血0,30和45 min组,每组又分为2亚组,其中对照组为胰岛细胞培养24 h;实验组为10 nmol/L Exendin-4与胰岛细胞培养24 h。应用双硫踪染色来计算分离胰岛数目并判断所提胰岛的纯度;丫啶橙/溴乙啶染色检测胰岛存活,胰岛素释放实验检测胰岛功能。结果与结论:随着热缺血时间的延长各组获得胰岛细胞的数量、纯度、存活率及功能逐渐降低。热缺血0 min组、30 min组、45 min组加入10 nmol/L Exendin-4培养24 h后分离、纯化的胰岛数量及纯度、活性均比未加入Exendin-4培养的对照组有所提高,热缺血30 min及热缺血45 min组中的实验组与对照组相比差异均有显著性意义(P<0.05)。提示Exendin-4对不同热缺血时间心死亡供体大鼠胰岛细胞具有保护作用,能减少胰岛细胞凋亡,改善胰岛的存活与功能;Exendin-4的使用可能成为胰岛移植早期缺血的边缘供体预处理的有效方法。BACKGROUND：The use of donor rat of cardiac death inevitably experiences warm ischemia injury, so the length of warm ischemia time plays a significant role on the number and function of pancreatic islet obtained. OBJECTIVE：To investigate the effect of Exendin-4 on pancreatic islet function of donor rats with cardiac death at different heat ischemia time. METHODS：Islet cells from Wistar rats were cultured in vitro and randomly divided into three groups according to the experimental conditions：0, 30, 45 min heat ischemia groups. Each group was further assigned into two subgroups, control group was cultured for 24 hours while experimental group wad cultured with 10 nmol/L Exendin-4 for 24 hours. The number of isolated pancreatic islets was calculated with diphenylthiocarbazone staining, and the purity of the extracted islets was adjusted. The viability of the islets was examined by AO/EB staining, and insulin secretion index assay was used to detect the function of the islets. RESULTS AND CONCLUSION：With the time of heat ischemia increasing, the number, purity, viability and function of islet cells obtained were decreased. After the cells in heat ischemia 0, 30, 45 min groups were cultured with 10 nmol/L Exendin-4 for 24 hours, the number, purity and viability of isolated and purified islets were increased compared to the group without added Exendin-4. There was significant difference between experimental group and control group in 30-minute and 45-minute ischemia groups （P〈0.05）. Exendin-4 can protect pancreatic islet cells in donor rats with cardiac death at different heat ischemia times, reduce the apoptosis, and improve islet survival and functions. The use of Exendin-4 can be an effective pretreatment method at early ischemia phase of islet transplantation.

关 键 词：实验动物 组织构建 胰岛细胞 热缺血时间 EXENDIN-4 胰岛移植 心死亡供体 

分 类 号：R318[医药卫生—生物医学工程]