目的:探讨人趋化素样因子1(chemokine-like factor 1,CKLF1)基因转移动员大鼠骨髓CD34^+细胞的作用和在心肌梗死情况下对外周血CD34^+细胞数量变化的影响。方法:肌肉电转不同剂量CKLF1质粒,构建大鼠急性心肌梗死模型,检测基因转移后不同时间外周血CD34^+细胞的数量,分析CD34^+细胞数与CKLF1质粒的量效关系和时间动力学的变化。结果:逆转录聚合酶链反应和免疫荧光染色均能检测到CKLF1的表达。CKLF1基因转移引起大鼠外周血CD34^+细胞的数量增加,在基因转移第5至7天达高峰,其中CKLF1质粒100μg组的峰值最高,分别为基因转移前的3.88倍和空质粒组的3.34倍(P〈0.01)。对心肌梗死大鼠CKLF1基因转移增加了心肌梗死后第1天外周血CD34^+细胞数的升高,其中100μg组最明显(14.61×10^6/L vs 7.85×10^6/L,P〈0.01)。结论:CKLF1能动员骨髓CD34^+细胞及在心肌梗死情况下增加外周血CD34^+细胞的数量,其中CKLF1质粒100μg效果最明显。
Objective: To assess the influence of different doses of CKLF1 plasmid on the dynamics and magnitude of the mobilization of the mobilization bone of marrow stem cells in a rat AMI model. Methods: Different doses of plasmid DNA encoding CKLF1 gene, empty plasmid or saline were injected into male SD rats intramuscularly with in vivo electroporation. Rats were subjected to left coronary artery ligation 6 days after gene transfer. Peripheral blood samples were drawn and CD34^+ cells were assayed by FACS calibur flow-cytometer. The changes in absolute number of CD34^+ cells were evaluated. Results: Expressions of CKLF1 mRNA and protein were detected in the injection site 7 days after gene transfer. Five days after gene transfer, the CD34^+ cells numbers in CKLF1 groups were significantly higher than those in empty plasmid group, especially in CKLF1100μg group (16.63 ×10^6/L vs 4.98 ×10^6/L, P 〈 0.01 ). On the 5 -7 days, the CD34^+ cell numbers in CKLF1 groups reached the peak and the peak number was 3.88 times that of baseline in CKLF1100 μg group ( P 〈 0.01 ). After AMI, the cell numbers of 1 day to 7 days were significantly higher than those of the baseline in empty plasmid group and saline group. In comparison to empty plasmid group, CKLF1 groups were associated with still higher numbers of cells 1 day after AMI ( P 〈0.05 ) , especially in CKLF1 100μg group ( 14.61×10^6/L vs 7.85 ×10^6/L, P 〈0.01 ). Conclusion: CKLF1 gene transfer significantly increases the mobilization of CD34^+ stem cells in acute myocardial infarction rats.