CAJ | 학술논문

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.
목적 탐토중약단분산B예처리적내피조세포(EPCs)대급성심기경사(AMI)대서이식골수간충질간세포(BMSCs)향심기분화조기기인표체적영향.방법 밀도제도리심법화첩벽사선법배양、순화EPCs여BMSCs;면역세포화학법(CD34/CD133/CD44)분별감정량충세포.결찰대서좌관상동맥,제작대서AMI모형,42지대서수궤분위가수술조,모형조,BMSCs조,EPCs+BMSCs조,기중치료조용단분산B최가약물농도처리적EPCs여BMSCs혼합,재대서AMI구주변분오점주사.RT-PCR검측심기분화조기기인Nkx2.5、GATA-4표체.결과 세포이식4 w후,EPCs+BMSCs조심기Nkx2.5、GATA-4표체량분별위2.256±0.524화1.567±0.287,여대조조비교차이현저.결론 단분산B예처리적EPCs연합BMSCs적이식상조료BMSCs향심기분화과정중Nkx2.5、GATA-4기인적표체,대심기세포적수량증가유중요영향,종이유가능개선심공능.
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.