骨髓间充质干细胞干预对急性肾损伤鼠肾脏中细胞因子的影响  

Effect of Mesenchymal Stem Cells on Cytokines in Kidneys of Acute Renal Injury Mice

作　　者：刘楠梅[1] 田军[1] 程劲[1] 胡大勇[1] 张金元[1] 

机构地区：[1]解放军第455医院肾内科,南京军区肾脏病专科中心,上海200052

出　　处：《中国中西医结合肾病杂志》2010年第4期310-313,I0006,共5页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：解放军南京军区医药卫生基金资助项目(No.07M023);南京军区122工程学科带头人培养基金资助项目;上海市卫生局科研计划资助项目(No.2009Y119)

摘　　要：目的:建立缺血再灌注(I/R)诱导的急性肾损伤小鼠模型,探讨外源性地给予骨髓间充质干细胞(MSCs)干预对模型小鼠肾损伤微环境中细胞因子水平的影响,并探讨其在肾损害修复中的作用。方法:采用Percoll密度梯度离心法分离出C57BL/6小鼠的骨髓间充质干细胞(mMSCs)。雄性C57BL/6小鼠45只,分为正常对照组(15只)、I/R组(15只、夹闭双侧肾蒂30min开放)、I/R+mMSCs组(15只、夹闭双侧肾蒂30min开放的同时尾静脉注射mMSCs)。于建模后1d、2d、3d、7d、14d分别处死部分小鼠(每次每组均处死3只),留取动脉血及肾组织,检测血尿素氮(BUN)及肌酐(Scr)水平,制作肾组织切片行HE染色,观察肾组织病理变化,行肾小管坏死程度评分(ATN评分)。ELISA法检测肾组织匀浆肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、单核细胞趋化蛋白-1(MCP-1)、白细胞介素-10(IL-10)、肝细胞生长因子(HGF)、骨形态发生蛋白-7(BMP-7)的水平。结果:I/R组小鼠的BUN及Scr均显著高于正常对照组,肾小管损伤严重;而I/R+mMSCs组小鼠的BUN及Scr水平较I/R组为低,以术后第7天差异最为显著(P<0.01),肾小管损伤病理明显减轻,ATN评分也有着显著降低。ELISA结果显示单纯I/R组各时间点肾组织匀浆中TNF-α、IL-1β、MCP-1的水平显著升高(P<0.01或P<0.05),而IL-10、HGF、BMP-7的水平却有着显著降低(P<0.01或P<0.05)。但同时给予MSCs干预的I/R小鼠肾组织匀浆中前述细胞因子的水平却向着相反方向改变。结论:MSCs对I/R肾组织中细胞因子的分泌具有调控作用,而这些调控了的细胞因子进而可通过旁分泌作用发挥促进肾损害的修复作用。Objective：Intervened by,To observe the effect of exogenous mesenchymal stem cells（MSCs）on cytokines among renal injury microenvironment in established mice model of acute renal ischemia/reperfusion injury.Methods：C57BL/6 mice＇s mMSCs had been successfully isolated by Percoll density gradient centrifugation.Total 45 healthy male C57BL/6 mice were divided into the control group（15）,the I/R group（15,clamping bilateral renal pedicles and then reopening after 30 minutes）,the I/R＋mMSCs group（15,clamping bilateral renal pedicles and then reopening after 30 minutes,meanwhile,mMSCs were injected through caudal vein into the body of model mice）.On 1 day,2 days,3 days,7 days,14 days after the model established,3 rats in each group were killed respectively,whose blood were sampled for detection of serum creatinine（Scr）and urea nitrogen（BUN）and kidneys for observation of the pathological changes with HE staining and evaluation of acute tubular necrosis score（ATN score）.The levels of tumor necrosis factor-α（TNF-α）,interleukin-1β（IL-1β）,monocyte chemoattractant protein-1（MCP-1）,interleukin-10（IL-10）,hepatocyte growth factor（HGF）,bone morphogenetic protein-7（BMP-7）in renal tissue homogenate were detected by enzyme-linked immunosorbentassay（ELISA）.Results：The levels of BUN and Scr in I/R group and I/R＋mMSCs group were much higher than those in the control group.While comparing with those in the I/R group,the levels of BUN and Scr in I/R＋mMSCs were much lower.At the same time,renal tubule＇s pathological changes in this group mitigated significantly and ATN score was also significantly lower.ELISA result showed that the levels of TNF-α,IL-1βand MCP-1 in I/R group＇s renal tissue homogenate were increased significantly（P〈0.01 or P〈0.05）,while the levels of IL-10,HGF and BMP-7 decreased notably.While mMSCs＇ intervention could change these cytokines levels to the opposite direction.Conclusion：MSCs can regulate cytokines secretion in renal tissue

关 键 词：骨髓间充质干细胞 缺血再灌注 细胞因子 

分 类 号：R692[医药卫生—泌尿科学]