α-1抗胰蛋白酶减轻人胰腺外分泌细胞对移植胰岛的损伤  

Protective effects of α-1 antitrypsin on human islets injury after transplantation by pancreas exocrine cells

作　　者：田磊[1] 周燕[2] 贾新干[1] 顾永耀[3] 吴向华[1] 李杰华[1] 

机构地区：[1]广西医科大学第一附属医院胃肠腺体外科,南宁530030 [2]广西医科大学药学院 [3]广西医科大学第一附属医院病理科,南宁530030

出　　处：《中华器官移植杂志》2010年第12期728-732,共5页Chinese Journal of Organ Transplantation

基　　金：广西科技厅青年基金（桂科青0640046、桂科青0728051）

摘　　要：目的 探讨胰腺外分泌细胞对胰岛移植物的损伤作用及α-1抗胰蛋白酶（A1AT）对胰岛β细胞的保护作用.方法 （1）体内实验：消化成人尸体部分胰腺,人工挑选胰岛,并收集胰腺外分泌细胞.链脲霉素腹腔注射诱导Balb/c-Nu裸小鼠成为糖尿病小鼠.对照组（n=6）小鼠每只于左肾包膜下移植成人胰岛250个;共同移植组（n=7）小鼠每只分别于左肾包膜上、下极同时植入胰岛250个和等体积的胰腺外分泌细胞.持续检测受鼠血糖,28d后切取左肾,以免疫组织化学染色（SP法）观察左肾包膜下抗淀粉酶抗体的表达,并继续检测血糖.（2）体外实验.纯化胰岛组（n=6）,每孔250个胰岛进行培养;非纯化胰岛组（n=6）,每孔250个胰岛和等体积外分泌细胞同时培养;非纯化胰岛＋A1AT组（n=6）,每孔250个胰岛和等体积外分泌细胞同时培养,并加入A1AT0.5 mg/ml.48 h后,测定各孔胰岛中胰岛素含量和上清液中胰蛋白酶浓度.结果 （1）胰岛移植后,对照组和共同移植组受鼠血糖均逐步恢复正常.共同移植组较对照组血糖恢复正常时间延迟,组间比较差异有统计学意义（P＜0.01）.切取受鼠左肾后5 d（移植后33 d）,两组受鼠血糖均＞21 mmol/L.共同移植组可见肾包膜下有大量抗淀粉酶抗体阳性细胞,对照组少见.（2）纯化胰岛组胰岛素含量为（2.02±0.33）μg/孔,非纯化胰岛组为（1.13±0.27）μg/孔,非纯化胰岛＋A1AT组为（1.68±0.17）μg/孔,各组间差异均有统计学意义（P＜0.01）.纯化胰岛组胰蛋白酶浓度为（1.03±0.25）ng/ml,非纯化胰岛组为（6.92±1.21）ng/ml,非纯化胰岛＋A1AT组为（3.23±0.55）ng/ml,各组间差异均有统计学意义（P＜0.01）.结论 胰腺外分泌细胞与胰岛同时移植会延迟植入胰岛功能恢复正常的时间,这与腺泡细胞内胰蛋白酶被激活、释放有关;在胰岛、胰腺外分泌细胞共同培养时加入A1AT,可以缓解腺泡细胞分�Objective To investigate the protective effects of α-1 antitrypsin on human islets injured by protease released from pancreas exocrine cells. Methods （ 1 ） in vivo experiment. Parts of the cadaveric pancreas was digested with collagenase, islets were selected artificially, and pancreatic exocrine cells were collected. 8-9 weeks olds male BALB/c-Nu nude mice were induced into diabetic mice with STZ （240 mg/kg body weight, i. p） and randomly divided into two groups： the control group （n = 6）, 250 islets were transplanted into left kidney subcapsule of diabetic nude mice; cotransplant group （n = 7）, 250 islets and the equal volume of pancreatic exocrine cells were transplanted into different regions of left kidney subcapsule. Blood glucose level was monitored. Nephrectomies were performed after 28 days. The expression of anti-amylase antibodies in subcapsule was detected by using immunohistochemical staining. （2） Islets culture： Three groups were randomly set up. Group 1： purified islet group, 250 islets were incubated into a 6-well culture plate; Group 2： non-purified islet group, 250 purified islets and equal volume of exocrine cells were incubated; Group 3： nonpurified islet ＋ Al AT group, 250 purified islets and equal volume of exocrine cells were incubated with α-1 antitrypsin added （0. 5 mg/ml）. After 48 h, insulin content of islets in each well and trypsin concentration in the supernatant of each well were measured. Results 10000 islets were collected.After islets transplantation, the blood glucose levels in control and co-transplant groups were normal,but a delayed islet function in reversing diabetes was in the co-transplant group, and ehe mice in both groups became hyperglycemic after nephrectomy. A large number of anti-amylase antibody-positive cells were found in renal subcapsule in the co-transplant group while little seen in the control group.Insulin levels in the non-purified islet group were decreased as compared with purified islet group,those in the non-purifie

关 键 词：胰岛移植 外分泌细胞 淀粉酶 α-1抗胰蛋白酶 

分 类 号：R686[医药卫生—骨科学]