糖尿病小鼠左肾被膜下胰岛细胞与胰腺外分泌细胞共同移植动物模型的建立

目的：建立糖尿病小鼠左肾被膜下同种异体胰岛细胞与胰腺外分泌细胞共同移植动物模型及探讨胰腺外分泌细胞对胰岛移植物的损伤作用.方法：（1）体内实验：采用胆总管内逆行灌注胶原酶联合淋巴细胞分离液的方法来分离纯化胰岛,人工挑取胰岛细胞并收集胰腺外分泌细胞.链脲佐菌素腹腔注射诱导BALB/C小鼠成为糖尿病小鼠.单纯移植组（n=10）每只小鼠于左肾被膜上极移植胰岛细胞250个,共同移植组（n =10）每只小鼠于左肾被膜上下极同时移植胰岛细胞250个和等体积的胰腺外分泌细胞,持续观测血糖及生命体征变化,1个月后切除左肾并继续检测血糖.（2）体外实验：利用双硫腙对胰岛进行特异性染色来计算胰岛产量及纯度,利用台盼蓝染色鉴定胰岛细胞的活性,以及用葡萄糖刺激胰岛素释放实验来检测胰岛功能.结果：（1）胰岛移植后,单纯移植组及共同移植组血糖均逐步降至正常,共同移植组较单纯移植组血糖恢复正常时间延迟,移植术后第2,3,4,5天,单纯移植组受鼠血糖低于共同移植组,差异有统计学意义（P＜0.05）.切除两组受鼠左肾3d后,两组受鼠血糖均＞21 mmol/L.（2）每只小鼠可获得150～200个高质量胰岛,纯度及活性均高于90％,葡萄糖刺激后胰岛素释放量明显增加（SI=2.90）.结论：（1）成功建立糖尿病小鼠左肾被膜下同种异体胰岛细胞与胰腺外分泌细胞共同移植动物模型.（2）胰腺外分泌细胞与胰岛细胞同时移植会延迟植入胰岛功能恢复正常的时间.

ESTABLISH OF ISLET CELL TRANSPLANTATION ANIMAL MODEL TOGETHER WITH THE PANCREATIC EXOCRINE CELLS UNDER THE LEFT RENAL CAPSULE IN DIABETIC MICE

Objective：To establish an animal model through islets and pancreatic exocrine cells transplantation under the left renal capsule of mice with allogeneic diabetes,and to investigate the damage of pancreatic exocrine cells of islets.Methods：（1） In vivo experiment：pancreas islet was isolated by perfusing the collagenase retrograde into the digestive pancreas in common bile duct,and purified with single density gradient centrifugation of lymphocyte separating medium.The islets and pancreatic exocrine cells were collected.Simple transplantation group （n =10）,250 islets were transplanted into left kidney subcapsule of diabetic mice,common transplantation group （n =10）,250 islets and the equaI 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.（2） In vitriol experiment：specific staining was conducted for the pancreas with dithizone to calculate the output and purification of pancreas islet.The activity of islet cells was identified with trypan blue staining,and the functions of pancreas islet were detected through stimulating the insulin release with glucose.Results：（1） After islet transplantation,the blood glucose levels in simple transplantation group and common transplantation group were normal,but a delayed islet function in reversing diabetes was in the common transplantation group,and each mice in both groups became hyperglycemic after nephrectomy.（2） High-quality pancreas islets （150 to 200） could be obtained from each mouse,and the purity and activity were above 90%,the insulin release amount after the stimulation from glucose increased obviously （SI=2.9）.Conclusion：（1） The animal model of islet and pancreatic exocrine cells transplantation under the left renal capsule in diabetic mice can be established successfully.（2） The pancreatic exocrine cells have detrimental effect on islet function after transplantation.