丹酚酸B干预EPCs对BMSCs移植的AMI大鼠心肌VEGF、bFGF蛋白表达的影响

目的]探讨中药丹酚酸B预处理的内皮祖细胞(EPCs)对骨髓间充质干细胞(BMSCs)移植后,急性心肌梗死(AMI)大鼠心肌血管新生的影响.(方法]密度梯度离心法和贴壁筛选法培养、纯化EPCs与BMSCs;免疫细胞化学法(CD34/CD133/CD44)分别鉴定两种细胞.结扎大鼠左冠状动脉,制作大鼠急性心肌梗死模型;丹酚酸B最佳药物浓度干预的EPCs,与BMSCs混合,在大鼠心肌梗死区周边分5点注射.免疫组织化学法检测心肌蛋白的表达.结果]细胞移植4周后,EPCs与BMSCs共移植组血管内皮生长因子(VEGF)、碱性成纤维细胞生长因子(bFGF)的积分光密度(IOD)分别13.179±3.053、23.634±4.705,与对照组差异具有统计学意义.结论]丹酚酸干预EPCs提高BMSCs移植后大鼠心肌VEGF、bFGF蛋白表达,有效改善了BMSCs向心肌分化的血管微环境.

Effects of EPCs preconditioned with salvianolic acid B combined with BMSC transplantation into infracted myocardium on expression of cardiac muscle VEGF and bFGF protein in rats

Objective] To investigate the effect of Salvianolic acid B-preconditioned endothelial progenitor cells (EPCs) combined withbone mesenchymal stem cells (BMSCs) on angiogenesis in rats with acute myocardial infarction (AMI).Methods] EPCs and BMSc werecultivated in vitro, purified by density-gradient centrifugation and plastic adherence method and identified by immunofluorescence andimmunocytochemistry (CD34/CD133/CD44).Acute myocardial infarction (AMI) was produced in rats by ligation of the coronary artery.EPCs preconditioned with Salvianolic acid B combined with BMSCs was injected into the five points of heart ischemic area.Four weekslater, we examined rat by ultrasonocardiograph, and the expression of Nkx2.5, GATA-4 by real-time PCR.Results] The IOD of VEGFand bFGF in EPCs and BMSCs group were 13.179±3.053, 23.634±4.705 respectively.It was increased remarkably comparing with thecontrol group.Conclusion] The EPCs preconditioned with Salvianolic acid B can improve the vascular microenvironment of BMSCstransplantation.