摘要
目的 观察与探讨生长抑素减轻小鼠胰岛移植过程中胰腺外分泌细胞对移植胰岛的损伤及其机制.方法 （1）体外实验：选取20只雄性BALB/C小鼠,采用随机数字表法分为生长抑素组和生理盐水组,每组10只.在获取胰岛手术麻醉前30 min,生长抑素组小鼠腹腔注射生长抑素10μg/g,生理盐水组注射等体积的生理盐水,然后分别收集两组小鼠的胰腺外分泌细胞及胰岛细胞,流式细胞仪检测两种细胞凋亡情况.（2）体内实验：获取小鼠胰岛及胰腺外分泌细胞,并对胰岛进行纯度及活性检测.应用链脲佐菌素建立小鼠糖尿病模型,然后进行胰岛移植,分别于小鼠左肾被膜上、下级同时移植胰岛250个及等体积的胰腺外分泌细胞.采用随机数字表法将成功建立胰岛移植模型的40只小鼠平均分为2组,实验组术后腹腔注射生长抑素,连续28 d,对照组术后注射等体积生理盐水,连续28 d.同时两组小鼠分别腹腔注射5乙炔基-2＆#39;-脱氧尿苷（EDU）5 μg/g,连续28 d.术后连续监测小鼠血糖;术后第8天,两组小鼠均进行葡萄糖耐量实验（IPGTT）;术后10和28 d两组各随机选取10只小鼠分别切除左肾,采用免疫组织化学法检测左肾包膜下抗淀粉酶抗体的表达,采用免疫荧光染色法检测移植胰岛β细胞增殖情况.结果 （1）流式细胞检测结果显示,生长抑素组小鼠胰腺外分泌细胞凋亡率明显高于对照组（P＜0.05）,两组间胰岛细胞凋亡率的差异无统计学意义（P＞0.05）;（2）获取的小鼠胰岛纯度及活性均高于95％.胰岛移植后,两组小鼠血糖均恢复正常,对照组较实验组血糖恢复正常时间延迟（P＜0.05）,对照组的血糖水平明显高于实验组;IPGTT实验结果显示移植物血糖调节功能正常;术后10d实验组和对照组移植物周围均可见抗淀粉酶抗体阳性颗粒,但实验组阳性颗粒数较对照组减少（P＜0.05）;免疫荧光检测显示,实验组�
Objective To investigate the protective effects of somatostatin （SS） on mice islets injury after transplantation by pancreas exocrine cells and its mechanism.Method （1） In vitro, 20 male BALB/C mice were randomly divided into the SS group （n =10） and the control group （n = 10）.The animals in SS group were injected with SS （10 g/g） by intraperitoneal injection （i.p） before 30 min, and those in the control group were given the same amount of normal saline （i.p）.The pancreas exocrine cells and islet cells in two groups were extracted respectively, and the apoptosis was detected by flow cytometric.（2） The pancreases of mice were digested with collagenase, islets and pancreatic exocrine cells were collected, and the purity and activity of islet was detected.In vivo, 8-9-week old male BALB/C mice were induced into diabetic mice with Streptozocin （STZ） （190 mg/kg body weight, i.p）.250 islets and the equal volume of pancreatic exocrine cells were transplanted into different regions of left kidney subcapsule.Forty mice were divided into two groups randomly.The experimental group was injected with SS （10 g/g, 3 times every day, i.p） for 28 days after operation, and the control group was injected with the same amount of normal saline （3 times every day, i.p） for 28 days.Then mice in two groups were injected with 5-Ethynyl-2＆#39;-deoxyuridine （EDU） （5 g/g, once every day, i.p） for 28 days.Blood glucose 1evel was monitored continually.Glucose tolerance test was performed after 8 days, and the left kidney was removed respectively after 10 days and 28 days.The expression of anti-amylase antibodies in subcapsule was detected by irnmunohistochemieal staining.The proliferation of islet beta cells was measured by immunofluorescence staining.Result （1） The apoptosis rate of pancreas exocrine cells in the experimental group was significantly higher than in the control group （P〈0.05）.There was no significant difference in the apoptosis rate of islet cells between the
出处
《中华器官移植杂志》
CAS
CSCD
2015年第7期417-423,共7页
Chinese Journal of Organ Transplantation
基金
教育部留学回国人员科研启动基金（教外司留[201131568号）
广西自然科学基金（2011GXNSFC018023）
广西医学科学实验中心开放基金专项项目（KFJJ2011-28）
广西卫生厅自筹经费科研项目（Z2010348）
广西高校科技研究立项项目（桂教201106LXl09）
关键词
胰岛移植
生长抑素
胰腺外分泌细胞
凋亡
Islet transplantation
Somatostatin
Pancreas exocrine cell
Apoptosis
