三种消化酶分离胰岛对移植胰岛存活的影响-第三军医大学学报.doc

三种消化酶分离胰岛对移植胰岛存活的影响 刘定志 杜成友* 罗诗樵 肖衡 潘龙 (重庆医科大学附属第一医院肝胆外科, 重庆400016) 摘要 目的：比较三种消化酶分离胰岛对细胞活力的影响，为临床胰岛移植消化酶的选取提供参考。 方法：相同条件下用Liberase TL、Collagenase P和CollagenaseⅤ消化胰腺，Ficoll-400密度梯度纯化获得胰岛，同基因胰岛肾被膜下移植，监测术后血糖变化和适时切取移植物HE染色观察组织学变化。 结果：成功分离、纯化小鼠胰岛细胞；每只受体可获得900±30个胰岛，3组受体均行肾被膜下移植；移植前各组小鼠血糖值比较无统计学意义（P0.05），术后A组动物血糖平均值水平较B、C组低，C组血糖平均值最高，各组间血糖值比较有统计学意义（P0.05）。HE染色：A组：胰岛形态完整，胞浆胞核区分明显，存活状态良好，炎症浸润不明显；B组：呈圆形或椭圆形，胞浆淡染，多核，细胞浸润较A组多；C组：炎症细胞浸润明显，伴胰岛破坏，形态尚规则。 结论：与Collagenase P和CollagenaseⅤ比较，Liberase TL消化小鼠胰腺可提高胰岛细胞团收获量、活性和功能。 关键词：胰岛移植 ；同基因 ；释放酶； 胶原酶；小鼠 The influence of the three digestive enzymes isolated from islet survival by transplanted islet LiuDingzhi,DuCheng you *, LuoShiqiao , XiaoHeng , PanLong (Department of Hepatobiliary surgery; the First Affiliated Hospital of Chongqing Medical University; Chongqing 400016; China) Abstract Objective：To compare three digestive enzymes isolated from islet and their effects on activity and provide the reference to Clinical islet transplantation for the selection of the digestive enzymes. Methods：Under the same conditions of pancreatic digestive enzymes, Liberase TL、Collagenase P and Collagenase Ⅴ. Ficoll-400 density gradient centrifugation was used for the extraction of islet. With syngenetic islet transplantation under renal capsule，monitoring the changes of blood sugar and timely cut out of graft Postoperatively, HE dyeing was used for the observation in histological changes.— Results: Successful isolation and purification of islets from pancreas in mice. Each receptor can obtain 900±30 islets. Three groups of receptors were adoption of renal capsule islet transplantation；Comparing the glucose levels for each mouse before transplantation is not significant（P0.05）. animal’s average blood sugar level in group A is low compared to group B and group C postoperatively，Group C is the highest. Comparison of each group’s blood sugar is statistically significant in value (P 0.05）. HE stain: Group A: Islet appearance is complete. Cytoplasm is c