骨髓间充质干细胞在肾缺血-再灌注损伤早期的抗凋亡作用

目的 观察骨髓间充质干细胞(mesenchymal stem cells,MSCs)自体移植对兔肾缺血-再灌注(ischemia reperfusion,I/R)损伤早期的治疗作用及其对肾小管上皮细胞凋亡的影响.方法 骨穿刺抽取新西兰大耳白兔股骨骨髓,分离、培养、扩增MSCs.30只新西兰大耳白兔随机分为移植组、对照组和假手术组,每组10只.移植组夹闭双肾动脉90 min,再灌注后经颈静脉移植BrdU标记的自体MSC8.对照组夹闭双肾动脉90 min,再灌注后经颈静脉注入等量生理盐水.假手术组假手术后经颈静脉移植Brdu标记的自体MSCs.肾I/R后48 h处死兔,比较兔肾功能和肾组织损伤,免疫组化法检测肾组织MSCs来源细胞,TUNEL法检测肾小管上皮细胞凋亡.用SPSS 13.0统计软件进行统计学分析,肾功能和肾小管细胞凋亡比较用ANOVA单因素方差分析;肾损伤指数比较用非参数的Kruskalwallis检验,以P<0.05为差异有统计学意义.结果 移植组较对照组肾功能明显改善(24 h,BUN:F=7.483,Scr:F=15.091;48 h,BUN:F=17.741,Scr:F=61.865;P<0.05),肾损伤明显减轻(F=12.658,P<0.01).移植组肾小管上皮细胞凋亡明显减少(F=135.495,P<0.01),且差异均具有统计学意义(P<0.05).I/R后48 h,移植组肾组织内可见少量BrdU阳性细胞.结论 MSCs自体移植对急性肾I/R损伤有治疗作用,其可能机制是通过减少肾小管上皮细胞凋亡来减轻肾I/R损伤早期的肾损害.

Anti-apoptosis role of bone marrow mesenchymal stem cells in the early phase of acute renal ischemia-reperfusion injury

Objective To evaluate the therapeutic effects of autolngous bone marrow mesenchymal stem cells transplantation in the early phase of acute renal ischemia-reperfusion(I/R)injury in rabbits and their impact on tubular epithelial cell apoptosis.Method Bone marrow mesenchymal stem cells(MSCs)were isolated and cultured in vitro.Thirty rabbits were divided randomly into transplanted group,control group and sham-operated group,10 in each group.Rabbits of transplanted group were induced acute renal I/R injury by clamping bilateral renal arteries for 90 minutes,then transplanted MSCs labeled with BrdU immediately after the resumption of circulation in kidney.The rabbits of control group were induced acute renal I/R injury and then infused with saline instead.The sharn operated rabbits were transplanted autologous MSCs labeled with BrdU after sham operation.Fortyeight hours after transplantation,all rabbits were sacrificed.Renal functional and structural damage were evaluated.MSCs derived BrdU positive cells were determined by immunostalning.Tubular epithelial cell apoptosis was arialyzed by usign TUNEL.The SPSS version 11.3 Software was used for statistical analysis.ANOVA were used to ana lyze the data of renal function and tubular epithelial cell apoptesis.Data of renal structure darnage were analyzed by using the nonparametric Kruskal-Wallis test.Results Compared with control group,the renal functional and struc tural darnage were significantly ameliorated(24 h,BUN:F=7.483,Ser:F:15.091;48 h,BUN:F:17.741.Scr:F=61.865;P<0.05),tubular epithelial cell apoptods Wsa reduced significantly in transplanted group(F=135.495,P<0.01).MSCs derived BrdU positive cell were detected in the renal tissues of transplanted rabbits.Conelusiu Autologous bone marrow mesenchymal stern cells transplantation is curative for acute renal I/R injury in rabbits.Bone marrow MSCs might ameliorate renal I/R injury by reducing tubular epithelial cell apoptosis.