血红素氧化酶1过表达对急性损伤肾脏中骨髓间充质干细胞的影响  

Effect of HO-1 overexpression on the survival of bone marrow-derived mesenchymal stem cells in the acute injured kidney

作　　者：刘楠梅[1] 王会玲[1] 韩国锋[1] 于秀峙[1] 田军[1] 胡伟锋[1] 张金元[1] 

机构地区：[1]中国人民解放军第455医院肾脏科南京军区肾脏病研究所,上海200052

出　　处：《肾脏病与透析肾移植杂志》2015年第5期441-446,共6页Chinese Journal of Nephrology,Dialysis & Transplantation

基　　金：上海市基础研究重大项目(12DJ1400203);国家自然科学基金青年项目(81300568);全军医学科技青年培育基金(13QNP050);上海市优秀青年医学人才培养计划基金(XYQ2013088);上海市青年科技启明星计划项目(12QA1405000)

摘　　要：目的:构建可高效表达血红素氧化酶1(HO-1)的大鼠骨髓间充质干细胞(BMSCs),观察HO-1过表达对移植BMSCs在缺血再灌注(I/R)诱导的急性肾损伤(AKI)肾脏中存活的影响,并探讨其机制。方法:基因转染技术获得HO-1-BMSCs和e GFP-BMSCs(空载体对照)。构建I/R诱导的AKI大鼠模型(I/R-AKI)并分别行BMSCs、e GFP-BMSCs和HO-1-BMSCs移植,移植3d后处死,检测大鼠肾功能及移植细胞在肾组织的分布。制作AKI的肾脏匀浆上清(AKI-KHS),体外干预培养的BMSCs、e GFP-BMSCs和HO-1-BMSCs,检测培养BMSCs的细胞凋亡、HO-1蛋白水平、氧化应激相关酶的活力及核因子κB(NF-κB)p65水平。结果:HO-1-BMSCs移植后,AKI大鼠肾脏中DAPI+细胞(BMSCs)比例最高,肾功能改善显著。经AKI-KHS干预后,培养BMSCs的TUNEL+细胞比例增加,HO-1过表达可显著逆转这一现象。AKI-KHS可诱导BMSCs细胞的HO-1表达增加,以HO-1-BMSCs/AKIKHS组升高最为显著,伴随着该组细胞内超氧化物歧化酶(SOD)和谷胱甘肽过氧化酶(GSH-Px)水平显著升高,丙二醛(MDA)和黄嘌呤氧化酶(XOD)活力降低,细胞内NF-κB p65核移位细胞比例也显著降低。结论:HO-1过表达可增强BMSCs在AKI肾脏微环境中的存活,HO-1的抗氧化作用及其下游NF-κB活化抑制为其可能的机制,期待可以解决移植干细胞在损伤靶器官中存活率低下的问题。Objective： HO-1-overexpressing bone marrow-derived mesenchymal stem cells（ HO-1-BMSCs） were constructed. Effect of HO-1 overexpression on the survival of BMSCs in the ischemia / reperfusion（ I / R）-induced injured kidney was observed,and the possible mechanism was also discussed. Methodology： HO-1-BMSCs and e GFP-BMSCs were obtained by the gene transfection technique. I / R-induced AKI rats were implanted with BMSCs. 3 days after implantation,the rats were sacrificed and the renal function and distribution of BMSCs in the nephridial tissues were measured. I / R-AKI kidney homogenate supernatant（ KHS） was harvested and used to culture BMSCs,e GFP-BMSCs and HO-1-BMSCs,respectively. The cell apoptosis,HO-1 expression,and level of NF-κB p65 were all tested respectively.Results： Proportion of DAPI＋BMSCs in the injured kidney was the highest in the HO-1-BMSCs implantation group together with the improved renal function of the AKI rats. The proportion of TUNEL＋cells were increased by AKI-KHS treatment in the cultured BMSCs,while HO-1 overexpression significantly inversed this phenomenon. AKI-KHS induced the increased expression of HO-1 in BMSCs,especially the highest in the HO-1-BMSCs. Furthermore,the activities of SOD and GSH-Px increased and the levels of MDA and XOD decreased significantly in the HO-1-BMSCs / AKI-KHS group. In addition,proportion of the NF-κB p65 nuclear translocation cells decreased significantly in this group. Conclusion： HO-1overexpression could enhance survival of BMSCs in the AKI microenvironment,the anti-oxidant effect of HO-1 with the inhibition of the activation of the downstream NF-κB is the possible mechanism. This finding is expected to solve the problem of the low living efficiency of the implanted cells in the injured organs.

关 键 词：血红素氧化酶1 骨髓间充质干细胞 急性肾损伤 氧化应激 核因子ΚB 

分 类 号：R692[医药卫生—泌尿科学]