芪丹液促进大鼠急性心肌梗死后心肌血管新生的机理研究

目的 观察益气活血复方芪丹液对大鼠急性心肌梗死后心肌血管新生的影响，并探讨其可能的分子机制。方法 采用大、中、小剂量芪丹液对Wistar大鼠冠状动脉左前降支结扎致急性心梗（acutemyocardial infarction，AMI）模型进行干预，并设正常组、假手术组、模型组和消心痛组。通过RT-PCR及免疫组化方法检测血管内皮生长因子（vascular endothelial growth factor，VEGF）及碱性成纤维细胞生长因子（basicfibroblastgrowthfactor，bFGF）基因及蛋白表达情况，利用免疫组化法测定Ⅷ因子，并计算缺血心肌微血管密度（microvascular density，MVD）。结果 模型组MVD较正常组及假手术组显著增加（P〈0、01）；消心痛及芪丹大、中剂量组结扎后7天、14天，其MVD均明显高于模型组（P〈0.05）。AMI后7天，模型组VEGF蛋白表达除较正常组升高外（P〈0．05），与其他组比较差异无显著性，14天时芪丹大、中剂量组及消心痛组VEGF表达均高于模型组（P〈0．05或P〈0、01）。AMI后7天、14天时模型组bFGF蛋白表达均上调，与正常组、假手术组比较差异有显著性（P〈0．01），且两时间点随时间延长表达下降（P〈0．05）；消心痛组及芪丹大剂量组7天及14天时bFGF表达均高于模型组（P〈0、01）。AMI后7天与14天模型组VEGFmRNA、bFGFmRNA均高于正常组、假手术组，消心痛及芪丹大、中、小剂量组高于模型组。各组7天与14天条带相比较，治疗组bFGFmRNA表达随时间的延长表达不变，而模型组缺血心肌中bFGFmRNA表达呈逐渐下降趋势。7天及14天VEGFmRNA的表达与时段变化没有明确的趋势。结论 芪丹液能够持续上调促血管新生因子VEGF、bFGF基因与蛋白的表达，提高缺血心肌毛细血管密度，促进心肌侧支循环的形成。

Acting Mechanism of Qidan Liquid in Promoting Myocardial Angiogenesis in Rats after Acute Myocardial Infarction

OBJECTIVE: To investigate the effect of Qidan Liquid (QL) on myocardial angiogenesis in rats after acute myocardial infarction (AMI) and explore its possible molecular mechanism. METHODS: AMI model was established by ligating the left anterior descending (LAD) coronary artery in Wistar rats, and intervened with QL in high, medium and low doses (QLh, QLm and QL1 group), and isosorbide dinitrate (ID group) respectively. Meanwhile, the model group, the sham group and the normal group were also set up. On the 7th (d7) and 14th day (d14) after modeling, mRNA and protein expressions of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) were detected with RT-PCR and immunohistochemistry, factor VIII expression in endothelial cells was examined, and microvascular density (MVD) in ischemic myocardium was calculated. RESULTS: MVD in the model rats increased significantly, higher than that in the normal and the sham group (P < 0.01), on d7 and d14, it was lower in the model group than that in the high and medium dose of QL groups and the ID group (P < 0.05). On d7, VEGF protein expression in the model group was higher than that in the normal group (P< 0.05), equivalent to that in the other groups, on d14, it was lower than that in the high and medium dose of QL groups and the ID group (P < 0.05 or P < 0.01). On d7 and d14, the protein expression of bFGF in the model rats increased significantly, higher than that in the normal and the sham group (P < 0.01), it decreased along the time progress between the two time points (P < 0.05), and lower than that in the high dose of QL group and ID group (P < 0.01). The mRNA expressions of VEGF and bFGF were higher in the model group than those in the normal and sham group, but lower than those in all the treated groups on d7 and d14. Comparison of the strips on d7 and d14 among various groups showed that the bFGF mRNA expression in the model group showed a descending trend as the time goes on, but in all the treated groups it was unchanged, while the change of VEGF mRNA expression didn't show any definite tendency. CONCLUSION: QL could up-regulate the protein and gene expressions of VEGF and bFGF persistently and increase MVD in ischemic myocardium to promote the development of collateral circulation in AMI rats.