兔自体骨髓间充质干细胞移植对冷缺血再灌注损伤肾脏细胞增殖与凋亡和肾功能的影响  

Effects of autologous bone marrow mesenchymal stem cells on the renal cell proliferation and apoptosis and renal function in a rabbit of renal ischemia-reperfusion injury

作　　者：方针强[1] 叶钢[1] 刘永亮[1] 周韬[1] 易善红[1] 邹旭东 

机构地区：[1]第三军医大学附属新桥医院泌尿外科、全军肾脏病中心,重庆400037 [2]贵州省习水县人民医院外一科,贵州564300

出　　处：《重庆医科大学学报》2009年第9期1166-1170,共5页Journal of Chongqing Medical University

基　　金：重庆市科委自然科学基金计划资助项目(CSTC,2009BB5154)

摘　　要：目的:建立兔肾脏冷缺血再灌注损伤(Ischemia-reperfusion injury,IRI)模型,观察经肾动脉灌注移植的自体骨髓间充质干细胞(BM-MSCs)对IRI肾脏细胞增殖与凋亡的影响,探讨其保护作用机制。方法:45只兔随机均分为假手术组(Ⅰ组)、对照组(Ⅱ组)和移植组(Ⅲ组)(n=15)。分离移植组兔BM-MSCs,建立肾脏IRI模型后,移植组于肾血流恢复后经肾动脉推注自体BM-MSCs,对照组和假手术组同法推注等量生理盐水。分别于术后3、7、14d3组各随机取5只经耳缘静脉采血,检测血清肌酐(Crea)和尿素(Urea),观察肾功能情况;同时取肾组织进行增殖细胞核抗原(PCNA)、Bcl-2、Bax免疫组化染色,TUNEL法检测肾脏凋亡细胞,计算Bcl-2、Bax含量和肾小管上皮细胞增殖指数(Proliferating cell nuclearantigen index,PI)和凋亡指数(Apoptosisindex,AI)。结果:IRI后第3d移植组和对照组兔Crea和Urea均达到最高水平[移植组:(297.9±18.6)μmol/L,(19.2±1.5)mmol/L;对照组:(326.6±20.0)μmol/L,(22.6±2.6)mmol/L],以后逐渐下降,到第14d,移植组Crea和Urea分别为(224.1±19.2)μmol/L,(13.9±1.6)mmol/L;对照组Crea和Urea分别为(278.6±16.9)μmol/L,(17.4±1.0)mmol/L,各时间点移植组Crea和Urea明显低于对照组(P<0.05)。各时间点移植组Bcl-2含量和肾小管上皮细胞PI高于对照组,Bax含量和肾小管上皮细胞AI低于对照组,两组间差异有统计学意义(P<0.05)。结论:BM-MSCs移植能促进肾脏I/R损伤后肾脏细胞增殖,抑制肾脏细胞凋亡,从而降低血清Crea和Urea,在一定程度上促进IRI后肾功能的恢复。Objective：To observe the protective effects of autologous bone marrow mesenchymal stem cells（BM-MSCs） on renal cell proliferation apoptosis during cold ischemia-reperfusion injury（IRI） in rabbit and to probe the mechanism of protective action on kidney. Methods：45 rabbits were divided randomly into three groups（n=15）： Sham operation group（ group Ⅰ）, control group of IRI（ group Ⅱ and BM-MSCs transplantation group（group Ⅲ）. After isolating the BM-MSCs of rabbits in group Ⅲ, to establish the rabbit models of renal IRI. Autologous BM-MSCs were infuded into rabbit in group Ⅲ, and the equivalent physiological saline in rabbit in group Ⅰ and Ⅱ. The detection was taken at the timepoints of 3,7 and 14 days after renal ischemia-reperfusion respectively. The serum levels of Crea and Urea were detected to evaluate the renal function, and Bax, Bcl-2 and PCNA in kidney were dectected with immunohistochemical staining. The apoptosis of cells was detected with TUNEL method. Results：The serum levels of Crea and Urea in group Ⅲ at different timepoint were lower than those in group Ⅱ significantly（P〈0.05）. The expressions of Bcl-2 protein and PI of renal tubular epithelia cells in group Ⅲat different timepoint were more than those in group Ⅱsignificantly（P〈0.05）and the expressions of Bax protein and AI of renal tubular epithelia cells in group Ⅲless than those in group Ⅱ significantly（P〈0.05）. Conclusions：BM-MSCs can promote cell proliferation and suppress apoptosis in kindey through up-regulating the expression of Bcl-2 protein and down-regulating the expression of Bax protein after renal IRI, thereby, decrease the serum levels of Crea and Urea, and promote the renal functional repair.

关 键 词：骨髓间充质干细胞 肾脏 缺血再灌注损伤 移植 

分 类 号：R365[医药卫生—病理学] R692[医药卫生—基础医学]