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目的观察粒细胞集落刺激因子(granulocyto-colony stimulating factor,G-CSF)对骨髓中内皮前体细胞(endothelial progenitor cells,EPCs)的动员作用,以及对心肌梗死交界区组织VEGF及其受体Flk-1mRNA表达的影响,并观察梗死交界区毛细血管密度的改善程度。方法雄性Wistar大鼠36只,体重250～280g,随机分为对照组、心肌梗死组(心梗组)和G-CSF组。心梗组和G-CSF组采用冠状动脉左前降支结扎法制作心肌梗死模型后,分别腹腔注射生理盐水(0.3mL/d)和G-CSF[30μg/(kg?d)],连续5d;对照组开胸后直接关胸,不行任何处理。给药后第7天球后静脉采血,测定EPCs水平和VEGF、C反应蛋白(C-reaction protein,CRP)浓度;RT-PCR检测心肌梗死交界区VEGF及其受体Flk-1mRNA的丰度。第4周采用Ⅷ因子免疫组织化学染色检测心肌梗死交界区毛细血管密度,行HE染色和Masson染色观察组织形态学变化。结果心梗组外周血白细胞数、EPCs水平以及VEGF和CRP浓度均较对照组增加(P<0.05),G-CSF组外周血白细胞数、EPCs水平以及VEGF浓度较心梗组进一步增高(P<0.05),但CRP浓度较心梗组有所降低(P<0.05)。心梗组VEGF及其受体Flk-1mRNA表达丰度均较对照组增强(P<0.05),G-CSF组较心梗组进一步增强(P<0.05)。G-CSF组较心梗组及对照组有更多的淋巴细胞浸润及新生毛细血管形成。结论 G-CSF能够动员骨髓中的EPCs进入外周血并上调梗死交界区VEGF及其受体Flk-1mRNA的表达丰度,增加心肌梗死交界区毛细血管密度,具有促进心肌梗死大鼠血管新生的作用。
Objective To observe the mobilization effect of granulocyto-colony stimulating factor (G-CSF) on endothelial progenitor cells (EPCs) in bone marrow and to investigate the effect of G-CSF on the expression of VEGF and its receptor Flk- 1mRNA expression, and to observe the improvement of capillary density in infarct border zone. Methods Thirty-six male Wistar rats weighing 250-280 g were randomly divided into control group, myocardial infarction group (MI group) and G-CSF group. The myocardial infarction group and the G-CSF group were injected with saline (0.3mL / d) and G-CSF [30μg / (kg? D)] intraperitoneally for 5 days after the left anterior descending coronary artery was ligated. Control group directly closed chest after thoracotomy, without any treatment. Blood samples were collected from the posterior vena cava on the 7th day after administration to determine the levels of EPCs and the levels of VEGF and C-reactive protein (CRP). The abundance of VEGF and its receptor Flk-1 mRNA in the border zone of myocardial infarction was detected by RT-PCR. At week 4, Ⅷ-factor immunohistochemical staining was used to detect the capillary density in the junctional area of myocardial infarction. HE staining and Masson staining were used to observe the histological changes. Results The numbers of leukocytes, EPCs and the levels of VEGF and CRP in myocardial infarction group were significantly higher than those in control group (P <0.05). The numbers of leukocytes, EPCs and VEGF in G-CSF group were higher than those in myocardial infarction group (P < 0.05), but CRP concentration was lower than the myocardial infarction group (P <0.05). The abundance of VEGF and its receptor Flk-1mRNA in myocardial infarction group were significantly higher than those in control group (P <0.05), and increased in G-CSF group compared with myocardial infarction group (P <0.05). The G-CSF group had more lymphocyte infiltration and neovascularization than the myocardial infarction group and the control group. Conclusion G-CSF can mobilize EPCs in bone marrow into the peripheral blood and upregulate the expression of VEGF and its receptor Flk-1 mRNA in infarct border zone, increase the density of capillary in the junctional area of myocardial infarction and promote angiogenesis in rats with myocardial infarction .