天津医药

• 实验研究 •    

SDF-1α/CXCR4信号通路在缺氧培养EPCs 治疗缺血性心脏病中的作用

简锴陶1,王联群1,王强1,杨东艳2,江力3,刘建实3   

  1. 1. 天津市胸科医院
    2. 天津胸科医院心外科
    3. 天津市胸科医院心外科
  • 收稿日期:2012-10-24 修回日期:2013-01-13 出版日期:2013-07-15 发布日期:2013-07-15
  • 通讯作者: 简锴陶

The Role of SDF-1 α/CXCR4 Signal Pathway in the Therapeutic Effects of Hypoxic Preconditioning of Endothelial Progenitor Cell Transplantation on Ischemia Heart Disease

JIAN Kai tao1,WANG Lian qun1,WANG Qiang 1,YANG Dongyan 1,JIANG Li 1,LIU Jianshi   

  • Received:2012-10-24 Revised:2013-01-13 Published:2013-07-15 Online:2013-07-15
  • Contact: JIAN Kai tao

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摘要：

【摘要】 目的  探讨基质细胞衍生因子-1α(SDF-1α)/CXCR4信号通路在缺氧培养骨髓内皮祖细胞(HEPCs)心肌内移植治疗缺血性心脏病中的生物学效应。方法  从同种系成年雄性Wistar大鼠长骨中获取骨髓内皮祖细胞(EPCs),常规培养4d后,1%O2+5%CO2+94%N2培养3d。观察HEPCs对细胞因子100μg/LSDF-1α的迁移能力变化,蛋白印迹法观察HEPCs表面SDF-1α受体CXCR4表达。将同种系成年雄性Wistar大鼠26只,随机分为对照组(n=8),常规组(n=9)和缺氧组(n=9),建立急性心肌梗死模型,在心肌梗死边缘5个不同区域心肌内分别注射PBS溶液200μL,EPCs2×106个,HEPCs2×106个,4周后用心脏超声分析血流动力学和心脏功能变化。结果  HEPCs与EPCs相比,细胞对SDF-1α迁移能力明显增强,细胞表面SDF-1α受体CXCR4表达增加。HEPCs心肌内移植4周后心脏超声示,与常规组和对照组比较,缺氧组左室收缩末期内径和射血分数得到明显改善(均P<0.05);与对照组比较,缺氧组和常规组左室舒张末期内径明显改善(均P<0.05);而缺氧组和常规组左室舒张末期内径比较,改善不明显(P>0.05)。结论 HEPCs上调细胞表面CXCR4,通过SDF-1α/CXCR4信号通路介导EPCs增强发挥细胞生物学功效。调节SDF-1α/CXCR4信号通路是优化EPCs移植治疗缺血性心脏病的有效方法。

关键词: 受体, CXCR4, 干细胞移植, 内皮细胞, 骨髓细胞, 心肌梗死, 缺氧, 趋化因子 CXCL12

Abstract: Abstract  Objective   We aimed at SDF-1α /CXCR4 signal pathway on hypoxic preconditioning endothelial progenitor cells(HEPCs), investigating the therapeutic effectiveness of transplantation HEPCs into acute infarcted myocardium Method    Bone marrow endothelial progenitor cells(EPCs) were islolated from syngeneic adult male Wistar rats, normoxic culture on EPCs for 4 days and 1%O2+5%CO2+94%N2 culture for 3 days. Using Boyden Chamber assay to compare migration ability between EPCs and HEPCs towards 100ng/ml bone marrow derived factor -1α(SDF-1α). Using western blot assay to detect expression of CXCR4, which is the solo receptor of SDF-1αon cells surface. Then, 26 syngeneic adult male Wistar rats were randomized into 3 groups: Control(n=8)，EPCs(n=9) and HEPCs(n=9),set up acute myocardium infarction animal model. At infarction, the rats received 5-points peri-infarct intramyocardial injections of PBS 200ul, 2×106 EPCs and 2×106 HEPCs. After 4 weeks, using ECHO to analyze improvement of cardiac function by haemodynamics parameters. Result    Compare with EPCs, the migration ability of HEPCs towards 100ng/ml SDF-1α was increased significant(549±78 cells/HPF vs 118±22 cells/HPF, p=0.009). The result of western blot of CXCR4 revealed that hypoxic preconditioning on EPCs upregulated CXCR4 expression on cell surface. After 4 weeks of transplantation, echocardiography of rats showed that compare HEPCs with EPCs and Control，systolic left ventricular internal diameter(LVIDs), ejection fraction(EF%) improved significantly. Compare HEPCs and EPCs with Control, diastolic left ventricular internal diameter(LVIDd) improved significantly, but there is no difference of LVIDd between HEPCs and EPCs. Conclusion    Regulating SDF-1α/CXCR4 pathway increased treatment effectiveness of cell transplantation and optimized therapeutic strategy on ischemia heart disease. Hypoxic preconditioning of EPCs upregulate CXCR4 expression on cell surface, improve migration ability of EPCs towards SDF-1α through SDF-1α/CXCR4 signal pathway. Transplantation of HEPCs after acute myocardium infarction limited left ventricular remodeling, improved cardiac function entirely.

Key words: receptors, CXCR4, Stem cell transplantation, endothelial cells, bone marrow cells, acute myocardiac infarction, anoxia, 趋化因子 CXCL12