丹酚酸B干预内皮祖细胞对骨髓间充质干细胞向心肌分化早期基因表.docVIP

丹酚酸B干预内皮祖细胞对骨髓间充质干细胞向心肌分化早期基因表 　 作者：李庆雯 谭俊珍 南亚昀 范英昌 【摘要】 目的 探讨中药丹酚酸B预处理的内皮祖细胞（EPCs）对急性心肌梗死(AMI)大鼠移植骨髓间充质干细胞（BMSCs）向心肌分化早期基因表达的影响。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AMI模型，42只大鼠随机分为假手术组，模型组，BMSCs组，EPCs+BMSCs组，其中治疗组用丹酚酸B最佳药物浓度处理的EPCs与BMSCs混合，在大鼠AMI区周边分五点注射。RTPCR检测心肌分化早期基因Nkx2.5、GATA4表达。结果 细胞移植4 w后，EPCs+BMSCs组心肌Nkx2.5、GATA4表达量分别为2.256±0.524和1.567±0.287，与对照组比较差异显著。结论 丹酚酸B预处理的EPCs联合BMSCs的移植上调了BMSCs向心肌分化过程中Nkx2.5、GATA4基因的表达，对心肌细胞的数量增加有重要影响，从而有可能改善心功能。 【关键词】 丹酚酸B；急性心肌梗死；内皮祖细胞；骨髓间充质干细胞；细胞移植 　　【Abstract】 Objective To investigate the effect of salvianolic acid B preconditioned endothelial progenitor cells combined with bone mesenchymal stem cells on gene expression in rats′ models of acute myocardial infarction(AMI). Methods Cultivation and purification of EPCs and BMSCs in vitro were by densitygradient centrifugation and plastic adherence method, and were identified by immunofluorescence and immunocytochemistry (CD34/CD133/CD44). The rat model of AMI was produced by ligation of the coronary artery. 42 rats were divided randomly by model, sham, BMSCs and EPCs combined with BMSCs groups, and EPCs preconditioned with salvianolic acid B combined with BMSCs were injected into the five points of heart ischemia area. After 4 weeks, the expression of Nkx2.5, GATA4 were examined by realtime PCR. Results The expression of NKx2.5 and GATA4mRNA were 1.700±0.382 and 1.135±0.257 respectively, which increased remarkably compared with control group. Conclusions The EPCs preconditioned with salvianolic acid B combined with BMSCs transplantation can increase the expression of NKx2.5 and GATA4, which improve the cardiac function. 　　【Key words】 Salvianolic acid B (SalB); Endothelial progenitor cells (EPCs); Bone mesenchymal stem Cells (BMSCs); Acute myocardialin infarction (AMI) 　　移植的干细胞能在受损心肌部位分化成为新的心肌样细胞〔1，2〕，而骨髓间充质干细胞（BMSCs）因自体来源，取材容易，扩增能力强，成为研究热点。目前研究主要集中在通过药物诱导来提高间充质干细胞（MSCs）移植后的存活和定向分化〔3，4〕。本文前期的研究结果表明丹酚酸B(SalB)可以减小心肌梗死的面积〔5〕,促进心脏功能的恢