血管内皮生长因子促进大鼠的移植胰岛再血管化并提高其存活率  

Influence of VEGF_(165) on graft islet revascularization and function

作　　者：程颖[1] 刘永锋[1] 张佳林[1] 李铁民[1] 赵宁[1] 

机构地区：[1]中国医科大学附属第一医院器官移植科,沈阳110001

出　　处：《中华器官移植杂志》2007年第6期332-335,共4页Chinese Journal of Organ Transplantation

基　　金：高等学校博士学科点专项科研基金(20050159010)

摘　　要：目的探讨血管内皮生长因子（VEGF）对大鼠的移植胰岛再血管化及对其存活率和功能的影响。方法构建质粒pIRES2-EGFP／VEGF165，以脂质体法在体外转染大鼠（供者）的血管内皮细胞。流式细胞术检测转染效率；免疫组织化学染色检测VEGF的表达。将糖尿病大鼠（受者）随机分为3组，每组10只。对照组：于肾被膜下单纯移植供者的300当量胰岛；实验组和空白转染组除移植供者的300当量胰岛外，分别加入供者的1×10^6个转染了质粒pIRES2-EGFP／vEGF165的血管内皮细胞和未经转染的正常血管内皮细胞。移植后监测受者的血糖及血清胰岛素水平。术后第14天，取受者肾脏，行HE染色及Insulin-6、VEGF和CD34免疫组织化学染色。结果血管内皮细胞中VEGFm的转染效率为13．06％。实验组供者的血管内皮细胞胞核和胞浆中均有VEGF表达。实验组受者于移植术后第3天血糖及胰岛素水平恢复正常；对照组和空白转染组虽有所改善，但未恢复到正常水平，与实验组相比，差异有统计学意义。实验组受者肾被膜下可见成团胰岛，Insulin-6免疫组织化学染色呈阳性，周围及内部有大量内皮细胞；VEGFm及CD34免疫组织化学染色呈阳性。对照组和空白转染组肾被膜下的细胞团中心细胞较少，部分被纤维组织代替，内部仅有少量CD34染色阳性的内皮细胞；Insulin-6免疫组织化学染色仅有少量细胞染成棕黄色；VEGF165免疫组织化学染色呈阴性。结论供者的胰岛与转染了VEGF165的血管内皮细胞共同移植给受者可以促进移植胰岛的再血管化，提高其存活率，并使其功能恢复正常。Objective To observe whether the elevated VEGF expression produced by the transfected vascular endothelial cells （VECs） could stimulate the angiogenesis of the grafts and its effect on the graft function. Methods The VECs were transfected by the pIRES2-EGFP plasmid, and then transplanted together with graft islets under the capsule of kidney. Flurocytometery was used to calculate the transfection efficiency. Immunohistochemical stain of human VEGF165 was used to detect the expression of VEGF in the VECs. The diabetic recipients were divided into three groups （n = 10 in each group） ： control group, 300 IEQ islets which considered as marginal grafts were transplanted under the capsule of the kidney; VEC group, the VECs were transplanted together with the islets; experirnental group, the VECs transfected by plRES2-EGFP/VEGF165 plasmid and the islets were transplanted. Blood glucose and insulin were determined every other day after operation. HE and immunohistochemical stain of Insulin-6, VEGF and CD34 for kidney was used to evaluate the histology of islet grafts. Results The transfected ratio was 13. 06% determined by flurocytometery. The positive cells of immunohistochemical stain of VEGF were observed in the experimental group. The blood glucose level and the plasma insulin level in the experimental group restored to the normal 3 days after transplantation. In contrast, diabetic rats receiving the same islets without or with normal VECs displayed moderate hyperglycemia, but there was no significant difference between the two groups. HE and immunohistochemical stain showed that large amount of islet grafts were seen under the capsule of the kidney, which were positively stained by insulin-6Ab and VEGF Ab. In the mass of the cells, the VECs positively stained by CD34 were observed. The similar mass was also seen in the two other groups, but with few positive cells stained by insulin-6 Ab and CD34 Ab. No positive cells stained by VEGF appeared in all groups. Conclusions Elevating the VEGF production by tran

关 键 词：基因转移技术 胰岛 移植 内皮细胞 

分 类 号：R657.5[医药卫生—外科学] R587.1[医药卫生—临床医学]