Tet-On调控NICD稳定表达的骨髓间充质干细胞治疗肾缺血再灌注损伤  

Bone marrow mesenchymal stem cells stably expressing NICD which is controlled by Tet-On system in the treatment of renal ischemia-reperfusion injury

作　　者：习小庆[1] 胡红林[1] 邹丛[2] 方周青 匡仁锐[1] 叶真逢[1] 黄雅为[1] Xi Xiao-qing;Hu Hong-lin;Zou Cong;Fang Zhou-qing;Kuang Ren-rui;Ye Zhen-feng;Huang Ya-wei(Department of Urology,the Second Affiliated Hospital of Nanchang University,Nanchang 330006,Jiangxi Province,China;Department of Endocrinology,the Fourth Affiliated Hospital of Nanchang University,Nanchang 330003,Jiangxi Province,China;Department of Surgery,the People’s Hospital of Pengze County,Jiujiang 332700,Jiangxi Province,China)

机构地区：[1]南昌大学第二附属医院泌尿外科,江西省南昌市330006 [2]南昌大学第四附属医院内分泌科,江西省南昌市330003 [3]江西省彭泽县人民医院外科,江西省九江市332700

出　　处：《中国组织工程研究》2018年第25期4035-4040,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81360110)~~

摘　　要：背景:肾缺血再灌注损伤是诱发急性肾损伤的主要原因。骨髓间充质干细胞治疗肾缺血再灌注损伤一直是研究的热点。目的:探讨Tet-On系统调控NICD稳定表达的骨髓间充质干细胞(NICD-Tet-BMSCs)对肾缺血再灌注损伤的修复作用。方法:采用密度梯度离心法分离培养大鼠骨髓间充质干细胞,体外构建Tet-On系统强力霉素调控NICD稳定表达的骨髓间充质干细胞,然后将此细胞移植至肾缺血再灌注损伤大鼠体内,术后检测血清肌酐水平、肾组织中骨髓间充质干细胞分布以及血管内皮生长因子和肿瘤坏死因子α表达变化。结果与结论:(1)NICD蛋白在NICD-Tet-BMSCs中稳定表达并受强力霉素调控;(2)与肾缺血再灌注损伤组相比,NICD-Tet-BMSCs组大鼠术后血清肌酐水平明显降低(P<0.05);(3)外源性骨髓间充质干细胞在NICD-Tet-BMSCs组大鼠肾组织中增殖持续时间更长;(4)NICD-Tet-BMSCs组肾组织中血管内皮生长因子蛋白表达升高,肿瘤坏死因子α蛋白表达降低,差异有显著性意义(P<0.05);(5)体外能够成功构建Tet-On系统调控NICD稳定表达的大鼠骨髓间充质干细胞,此细胞可促进大鼠肾缺血再灌注损伤的修复。BACKGROUND:Renal ischemia-reperfusion injury(IRI)is the main cause of acute kidney injury(AKI).The treatment of renal IRI by bone marrow mesenchymal stem cells(MSC)has always been an issue of concern.OBJECTIVE:To construct BMSCs stably expressing NICD controlled by Tet-On system in vitro(NICD-Tet-BMSCs),and to explore its therapeutic effect on renal IRI.METHODS:We separated and cultured rat BMSCs by density gradient centrifugation,and constructed NICD-Tet-BMSCs in vitro.The cells were then transplanted to renal IRI rats.Serum creatinine level,the distribution of BMSCs in renal tissue and the expression of vascular endothelial growth factor and tumor necrosis factorαin the kidney were detected thereafter.RESULTS AND CONCLUSION:NICD protein stably expressed in the NICD-Tet-BMSCs which was regulated by the doxycycline.Compared with the control group,the renal IRI rats undergoing NICD-Tet-BMSCs transplantation had significantly lower serum creatinine levels(P<0.05),sustained cell proliferation,increased vascular endothelial growth factor protein expression and decreased tumor necrosis factorαprotein expression in the kidney(P<0.05).Therefore,NICD-Tet-BMSCs can be successfully established in vitro,and promote the reparation of renal IRI in rats.

关 键 词：骨髓间充质干细胞 NICD TET-ON 强力霉素 肾缺血再灌注损伤 肌酐 血管内皮生长因子 肿瘤坏死因子α 国家自然科学基金 骨髓 间质干细胞 肾 再灌注损伤 组织工程 

分 类 号：R394.2[医药卫生—医学遗传学]