α-1抗胰蛋白酶减轻小鼠胰腺外分泌细胞对移植胰岛的损伤  

A1AT alleviates pancreas exocrine cells damage to transplanted islets in mice

作　　者：韩小乐[1] 黎沙 夏小林[1] 糜亮亮 周乐亮[1] 田磊[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530021 [2]广西医科大学附属肿瘤医院

出　　处：《中华器官移植杂志》2015年第2期102-107,共6页Chinese Journal of Organ Transplantation

基　　金：教育部留学回国人员科研启动基金，广西自然科学基金，广西医学科学实验中心开放基金专项，广西研究生创新计划

摘　　要：目的探讨α-1抗胰蛋白酶（A1AT）减轻胰腺外分泌细胞对移植胰岛的损伤及促进胰岛口细胞增殖。方法（1）体外实验：选10只BALB／c小鼠，挑取所有的胰岛并收集胰腺外分泌细胞。纯化胰岛组（n=6），每孔100个胰岛，常规培养；实验组（n=6），每孔100个胰岛和等体积外分泌细胞同时培养，并加入A1AT（0．5mg／m1）；对照组（n=6），每孔100个胰岛和等体积外分泌细胞同时培养。48h后，收集各孔上清液，分别检测胰岛素含量及胰蛋白酶浓度；然后分别换低糖、高糖RPMI-1640培养液，行胰岛素释放实验。（2）体内实验：链脲佐菌素腹腔注射诱导BALB／c小鼠成为糖尿病小鼠。实验组（n=10）和对照组（n=10）分别于小鼠左肾被膜上下级同时移植胰岛细胞250个和等体积的胰腺外分泌细胞，术后实验组按腹腔注射A1AT83mg／kg（28d），对照组注射等量生理盐水（28d），同时两组小鼠分别按0．5btg／g腹腔注射EDU（28d）。1个月后切除左肾，免疫组织化学染色法检测左肾包膜下抗淀粉酶抗体的表达，免疫荧光染色法检测移植胰岛B细胞增殖情况，并继续检测血糖。结果（1）纯化胰岛组胰岛素含量为（1．53±0．33）μg／孔，实验组为（1．26±0．13）μg／孔，对照组为（0．82±0．24）μg／孔，各组间的差异均有统计学意义（P〈0．01）。纯化胰岛组胰蛋白酶浓度为（1．05±0．22）ng／ml，实验组为（2．42±0．44）ng／ml，对照组为（5．73±1．12）ng／ml，各组间的差异均有统计学意义（P〈0．01）。纯化胰岛组胰岛素刺激指数为4．11±1．45，实验组为2．53±1．52，对照组为1．32±1．12，各组间的差异均有统计学意义（P〈0．01）。（2）胰岛移植后，实验组和对照组血糖均降至正常，对照组较实验组血糖恢复正常时间延迟，组间差异有统计学意义（P〈0．05）。切除两组小鼠左肾3Objective To investigate the effect of alpha 1-antitrypsin （A1AT） concerning in reducing the injury of transplanted islets by pancreas exocrine cells and promoting proliferation of the pancreas B cells. Method The pancreases of mice were digested with eollagenase, islets were isolated artificially, and pancreatic exocrine cells were collected. In purified islet group （n = 6）, 100 islets were seeded into a 6-well culture plate. In experimental group（n = 6）, 100 islets were co-cultured with equal volume of pancreas exocrine cells, and 0. 5 mg/mL A1AT was added into a 6-well culture plate. In control group（n = 6）, 100 islets were co-cultured with equal volume of pancreas exocrine cells. After 48 h, insulin content of islets in each well and trypsin concentration in the supernatant of each well were measured. The islets were cultured in low sugar and high sugar 1640 medium, then glucose stimulated insulin secretion （GSIS） test was carried out. In vivo, 8-9-week old male BALB/C mice were induced with STZ （190 mg/kg body weight, i. p） to establish the diabetic model and randomly divided into two groups. In experimental（n = 10） and control（n = 10） groups, 250 islets and the equal volume of pancreatic exocrine cells were transplanted into different regions of left kidney subcapsule, resepctively. The experimental group was injected with A1AT （83 mg/kg, qd, i. p） for 28 days after operation, and the control group was injected with the same amount of normal saline （qd, i. p） for 28 days. Both two groups were given EDU （5μg/g, qd, i. p） for 28 days. The blood glucose level was monitored continually. Nephrectomies were performed after 28 days. The expression of anti-amylase antibodies in the renal subcapsule was detected by immunohistochemical staining, and the proliferation of islet beta cells was examined using immunofluorescence staining. Result Insulin levels and insulin stimulation index in the control group were decreased as compared with those in the purified islet group; t

关 键 词：胰岛移植 胰腺外分泌细胞 胰蛋白酶 

分 类 号：R657.5[医药卫生—外科学]