肾被膜下胰岛移植治疗1型糖尿病模型小鼠的研究  

Treatment of Type 1 Diabetes Model Mice with Islet Transplantation under Renal Capsule

作　　者：刘森峰 冯樾 王圣禹 冯逸飞 莫庆辉 田磊 LIU Sen-feng;FENG Yue;WANG Sheng-yu;FENG Yi-fei;MO Qing-hui;TIAN Lei(Gastrointestinal Gland Surgery,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,Guangxi,China)

机构地区：[1]广医科大学第一附属医院胃肠腺体外科,广西南宁530021

出　　处：《医学信息》2021年第4期71-74,共4页Journal of Medical Information

基　　金：国家自然科学基金地区科学基金项目(编号:81660134);广西自然科学基金面上项目(编号:2017GXNSFAA198051、2016GXNSFAA380162)。

摘　　要：目的观察经肾被膜下小鼠胰岛移植模型改良方法的移植效果。方法采用胆总管内逆行灌注联合胰腺离体多点灌注法消化胰腺,镜下手工挑选胰岛细胞,分别用台盼蓝染色检测活性,双硫腙(DTZ)染色检测纯度,葡萄糖刺激胰岛素释放实验检测分离提取胰岛的功能。链脲佐菌素170 mg/kg腹腔内注射,建立1型糖尿病小鼠模型;空白对照组(n=13),实验组(n=13)为1型糖尿病模型小鼠并给予同种异体胰岛细胞肾被膜下移植,移植术后监测随机血糖,移植后第10天行葡萄糖耐量检测,移植后第29天切除实验组左肾行HE染色观察移植物情况。结果每只小鼠可以获得(200.23±30.18)个胰岛细胞团,且纯度和活性都>90%;高糖组每个胰岛素释放量为(1.98±0.20)ng/胰岛,低糖组检测每个胰岛素释放量为(0.69±0.11)ng/胰岛,高糖组为低糖组的2.87倍(P<0.05);移植术后实验组血糖第1、2天逐步下降,并在第3天全部降至正常,移植术后第4、6、7、14、21、28天两组血糖水平比较,差异无统计学意义(P>0.05);实验组摘除左肾术后血糖比较,差异有统计学意义(P<0.05);两组糖耐量比较,差异无统计学意义(P>0.05);实验组切除左肾后血糖持续升高,HE染色切片显示肾被膜下胰岛移植物表面和内部已有新生血管生成存活良好。结论采用改进后分离和移植胰岛方法可以保证获得高质量的胰岛细胞,提高胰岛产量及移植模型的成功率,是一种高效稳定的技术方法,为下一步实验药物干预筛选奠定了基础。Objective To Observe the transplantation effect of the modified method of mouse pancreatic islet transplantation under the renal capsule.Methods The pancreas was digested by retrograde perfusion in the common bile duct combined with multi-point perfusion of the pancreas in vitro,the islet cells were manually selected under the microscope,and the activity was detected by trypan blue staining,the purity by dithizone(DTZ)staining,and the glucose-stimulated insulin release testfunction of separation and extraction of islets.Streptozotocin 170 mg/kg was injected intraperitoneally to establish a mouse model of type 1 diabetes;blank control group(n=13),the experimental group(n=13)was type 1 diabetes model mice and received allogeneic islet cell transplantation under the renal capsule.Random blood glucose was monitored after transplantation,glucose tolerance was tested on the 10th day after transplantation,and the left kidney of the experimental group was removed on the 29th day after transplantation to observe the graft status.Results Each mouse could obtain(200.23±30.18)islet cell clusters,and the purity and activity were both greater than 90%;The amount of insulin released per insulin in the high glucose group was(1.98±0.20)ng/islet,and the amount of insulin released in the low glucose group was(0.69±0.11)ng/islet,and the high glucose group was 2.87 times that of the low glucose group(P<0.05);the blood glucose of the experimental group decreased gradually on the first and second days after transplantation,and all fell to normal on the third day.The blood glucose levels of the two groups were compared on the 4,6,7,14,21,and 28 days after transplantation,the difference was not statistically significant(P>0.05);The blood glucose after removal of the left kidney in the experimental group was significantly different(P<0.05);There was no statistically significant difference in glucose tolerance between the two groups(P>0.05);In the experimental group,blood glucose continued to rise after the left kidney was removed,and H

关 键 词：胆总管逆行灌注 胰腺离体多点灌注 胰岛移植 动物模型 低位肾被膜开口 

分 类 号：R335[医药卫生—人体生理学]