转化生长因子β1趋化骨髓间充质干细胞修复缺血再灌注肾损伤的机制研究  

Mechanism of Transforming Growth Factor-β1 on Migration of Bone Marrow Mesenchymal Stem Cells for Repairing Renal Ischemia Reperfusion Injury

作　　者：徐申 司晓芸[1] 毕晓红[1] 吴小燕[1] XU Shen;SI Xiaoyun;BI Xiaohong;WU Xiaoyan(Department of Nephrology,Zhongnan Hospital of Wuhan University,Wuhan Hubei 430071,China)

机构地区：[1]武汉大学中南医院肾内科,湖北武汉430071

出　　处：《华南国防医学杂志》2018年第7期440-445,共6页Military Medical Journal of South China

基　　金：湖北省自然科学基金资助项目(2009CDB428;2015CFB407)

摘　　要：目的探讨转化生长因子β1(transforming growth factor-β1,TGF-β1)趋化骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)修复大鼠肾缺血再灌注损伤(ischemia reperfusion injury,IRI)的可能机制。方法制作大鼠缺血再灌注肾损伤匀浆,与BMSCs共培养。分为4组:BMSCs组、匀浆组、TGF-β1中和抗体组和CXC趋化因子受体4(CXC chemokine receptor 4,CXCR4)中和抗体组。免疫荧光染色与激光共聚焦显微镜检测BMSCs表面CXCR4受体表达;体外跨膜趋化实验观察BMSCs向基质细胞衍生因子1(stromal cells derived factor-1,SDF-1)的迁移能力;实时荧光定量聚合酶链式反应(quantitative real-time-polymerase chain reaction,q PCR)法检测SDF-1α、CXCR4、TGF-β1 mRNA表达。结果BMSCs表面CXCR4受体低水平表达,匀浆组CXCR4表达明显增强,增强的表达可被TGF-β1抗体和CXCR4抗体所抑制。匀浆组BMSCs迁移细胞数[(38.92±6.79)个/cm^2]明显增加,显著高于BMSCs组[(18.47±5.02)个/cm^2]、TGF-β1中和抗体组[(13.26±4.03)个/cm^2]、CXCR4中和抗体组[(7.03±2.14)个/cm^2],3组分别与匀浆组比较差异具有统计学意义(t分别为12.94,15.33,21.86,P<0.01)。匀浆组SDF-1αmRNA表达为(1.92±0.36),TGF-β1中和抗体组(1.71±0.28),CXCR4中和抗体组(1.66±0.29),3组间SDF-1αmRNA表达比较差异无统计学意义(F=3.16,P>0.05)。匀浆组CXCR4 mRNA表达(3.44±0.62)显著上调,显著高于BMSCs组(1.36±0.57)、TGF-β1中和抗体组(1.82±0.48)、CXCR4中和抗体组(1.15±0.51),3组分别与匀浆组比较差异具有统计学意义(t分别为8.72,10.93,13.27,P<0.01)。匀浆组TGF-β1 mRNA(2.91±0.72)表达显著上调,明显高于TGF-β1中和抗体组(0.52±0.21),组间比较差异具有统计学意义(t=18.35,P<0.01),但CXCR4中和抗体组(2.77±0.83)与匀浆组比较差异无统计学意义(t=0.52,P>0.05)。结论缺血再灌注肾损伤促进TGF-β1分泌,高表达的TGF-β1通过提高BMSCs表面CXCR4受体表达诱导BMSCs迁移修复大鼠肾IRI。Objective To explore the possible mechanism of transforming growth factor-β1（ TGF-β1） on migration of bone marrow mesenchymal stem cells（ BMSCs） for repairing renal ischemia reperfusion injury（ IRI） in rats. Methods The BMSCs were co-cultured with IRI renal homogenate supernatant. Cultured BMSCs were divided into four groups： BMSCs group,homogenate group,TGF-β1 neutralize antibody group and CXC chemokine receptor 4（ CXCR4） neutralize antibody group. The expression of CXCR4 receptor on BMSCs surface was observed by immuno-fluorescence staining and laser focus microscope. The migration ability of BMSCs to stromal cells derived factor-1（ SDF-1） was investigated by Transwell test in vitro,and SDF-1 α,CXCR4 and TGF-β1 mRNA expressions were detected by quantitative real-time-polymerase chain reaction（ q PCR）. Results CXCR4 receptor was expressed at a low level on BMSCs surface and significantly increased in homogenate group,meanwhile,the increased expression could be inhibited by TGF-β1 and CXCR4 antibodies.The cells numbers on migration of BMSCs in homogenate group were significantly higher than those in BMSCs group,TGF-β1 neutralize antibody group and CXCR4 neutralize antibody group [（ 38. 92 ± 6. 79） cells/cm^2 vs.（ 18. 47 ± 5. 02）cells/cm^2,（ 13. 26 ± 4. 03） cells/cm^2,（ 7. 03 ± 2. 14） cells/cm^2,t = 12. 94,15. 33,21. 86,respectively,P〈0. 01]. SDF-1 α mRNA expressions in homogenate group,TGF-β1 neutralize antibody group and CXCR4 neutralize antibody group were 1. 92± 0. 36,1. 71 ± 0. 28,1. 66 ± 0. 29,respectively（ F = 3. 16,P〈0. 05）. CXCR4 mRNA expressions in homogenate group were significantly up-regulated and higher than those in BMSCs group,TGF-β1 neutralize antibody group and CXCR4 neutralize antibody group（ 3. 44 ± 0. 62 vs. 1. 36 ± 0. 57,1. 82 ± 0. 48,1. 15 ± 0. 51,t = 8. 72,10. 93,13. 27,respectively,P〈0. 01）. TGF-β1 mRNA expressions in homogenate group were markedly up-regulated and higher than those in TGF-β1 neutralize antibody

关 键 词：转化生长因子Β1 缺血再灌注损伤 骨髓间充质干细胞 CXC趋化因子受体4 

分 类 号：R332[医药卫生—人体生理学] R692[医药卫生—基础医学]