骨髓间充质干细胞表面CXCR4过表达影响缺血-再灌注大鼠肾损伤修复的机制研究  

Effects of mesenchymal stem cells surface CXC chemokine receptor 4 over-expression on repair of ischemia reperfusion injured kidney

作　　者：司晓芸[1] 李静静[1] 毕晓红[1] 吴小燕[1] 

机构地区：[1]武汉大学中南医院肾内科,武汉430071

出　　处：《中华急诊医学杂志》2015年第12期1390-1395,共6页Chinese Journal of Emergency Medicine

基　　金：湖北省自然科学基金（2009CDB428,2015CFB407）

摘　　要：目的 观察骨髓间充质干细胞（MSCs）表面CXCR4受体过表达对大鼠缺血-再灌注（I/R）肾损伤修复的影响.方法 制备I/R肾损伤匀浆,与MSCs共培养,Western blot检测细胞基质细胞衍生因子-1 （SDF-1α）和CXCR4膜蛋白表达,Transwell实验观测匀浆处理后MSCs向SDF-1迁移的能力.建立I/R肾损伤模型,观察正常对照组、I/R肾损伤组、MSCs输注组和CXCR4中和抗体干预组病理改变;免疫荧光组织化学法检测肾组织CXCR4蛋白表达;Real-time定量PCR法检测SDF-1α、CXCR4、肝细胞生长因子（HGF）和表皮生长因子（EGF） mRNA表达.多组间比较采用单因素方差分析,两组间比较采用独立样本t检验.结果 SDF-1α蛋白表达在I/R肾损伤匀浆组明显升高,但与CXCR4抗体组间比较差异无统计学意义（t=0.862,P=0.403）,L/R肾损伤匀浆能够显著上调MSCs CXCR4蛋白表达（F=95.957,t=10.166,P＜0.01）,并增强其向SDF-1迁移的能力（F=82.459,t=6.826,P＜0.01）,CXCR4中和抗体能够显著下调CXCR4蛋白表达（t=13.657,P＜0.01）和抑制迁移（t=12.662,P＜0.01）.I/R肾损伤组大鼠肾皮质肾小管结构破坏,MSCs移植能够改善损伤的肾小管,但修复作用可被CXCR4抗体所抑制.I/R肾损伤组SDF-1α蛋白和mRNA表达升高,但组间差异无统计学意义（F=1.909,P=0.173）.MSCs移植能够显著上调CXCR4蛋白和mRNA表达（F=6.663,P=0.006）,随CXCR4表达增加,HGF（F=11.898,P＜0.01）和EGF mRNA表达逐渐升高（F=5.309,P＜0.05）,CXCR4中和抗体能够下调该表达（t=5.312,=4.310,P＜0.01）.结论 I/R肾损伤时,MSCs表面CXCR4受体表达升高,能够促进MSCs迁移,并分泌肾保护性细胞因子促进肾损伤修复.Objective To observe the effects of mesenchymal stem cells （MSCs） surface CXC chemokine receptor 4 over-expression on the repair of kidney after ischemia reperfusion （I/R） injury.Methods The MSCs were co-cultured with I/R injured renal cell homogenate supernatant.The MSCs surface CXCR4 and stromal cells derived factor-1 （SDF-1α） protein levels were detected by Western blot, chemotactic ability of MSCs to SDF-1 was investigated by transwell test.The I/R injured renal model was made and pathological changes were observed in control group, I/R group, MSCs injection group and CXCR4 neutralize antibody group.Renal CXCR4 protein expression was measured by immunofluorescence histochemistry, SDF-1α、 CXCR4、 hepatocyte growth factor （HGF） and epidermal growth factor （EGF） mRNA were detected by Real-time quantitative polymerase chain reaction （RT-PCR）.Comparisons among multiple groups were performed using One-way analysis of variance, and comparisons between groups were carried out using independent-sample t-test.Results In vitro, the SDF-1α protein expression markedly increased in I/R injured renal tissue homogenate, but the difference was not significant between I/R group and CXCR4 antibody group （t =0.862, P =0.403）.MSCs surface CXCR4 protein expression increased significantly after co-cultured with I/R injured renal tissue homogenate （F =95.957, t =10.166, P 〈 0.01）, and the chemotactic ability of MSCs to SDF-1 increased at the same time （F =82.459, t =6.826, P 〈 0.01）, the CXCR4 protein expression （t =13.657, P 〈 0.01） and the chemotactic ability （t =12.662, P 〈0.01） could be decreased by CXCR4 neutralize antibody.In vivo, renal tubular structure was destroyed in I/R group.After MSCs injection, the renal pathological injury improved rapidly, but the improvement could be inhibited by CXCR4 antibody.The expression of SDF-1α mRNA and level of SDF-1a protein increased in I/R group, but there was no significant difference among different groups （F =1.909,P =0.173

关 键 词：间充质干细胞 CXCR4受体 缺血-再灌注 大鼠 肾损伤 迁移 旁分泌 修复 

分 类 号：R692[医药卫生—泌尿科学]