CAJ | 학술논문

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。
배경：심사망공체대서공이적이용불가피면지경과열결혈손상，인차열결혈시간적장단대획득적이도수량급공능유중요영향。목적：관찰Exendin-4대불동열결혈시간심사망공체대서이도공능적영향。방법：대서이도세포체외배양，근거실험조건수궤분위3대조：열결혈0，30화45 min조，매조우분위2아조，기중대조조위이도세포배양24 h；실험조위10 nmol/L Exendin-4여이도세포배양24 h。응용쌍류종염색래계산분리이도수목병판단소제이도적순도；아정등/추을정염색검측이도존활，이도소석방실험검측이도공능。결과여결론：수착열결혈시간적연장각조획득이도세포적수량、순도、존활솔급공능축점강저。열결혈0 min조、30 min조、45 min조가입10 nmol/L Exendin-4배양24 h후분리、순화적이도수량급순도、활성균비미가입Exendin-4배양적대조조유소제고，열결혈30 min급열결혈45 min조중적실험조여대조조상비차이균유현저성의의(P<0.05)。제시Exendin-4대불동열결혈시간심사망공체대서이도세포구유보호작용，능감소이도세포조망，개선이도적존활여공능；Exendin-4적사용가능성위이도이식조기결혈적변연공체예처리적유효방법。
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.