抗内皮细胞黏附分子-1靶向微泡介导hAng-1基因转染治疗兔心肌缺血的实验研究

Targeted anti-ICAM-1 microbubbles mediated hAng-1 gene transfection in treatment of ischemic myocardium in rabbits

目的探讨应用抗心肌内皮细胞粘附分子-1(ICAM-1)靶向微泡能否提高血管生成素-1基因在缺血心肌中的转染效率。方法体外实验检测抗ICAM-1靶向微泡与经人白细胞介素-1β刺激后的ECV304细胞结合的程度。体内实验分三组:①对照组:hAng-1基因+超声照射;②非靶向微泡组:携hAng-1基因微泡+超声照射;③靶向微泡组:携hAng-1基因的抗ICAM-1靶向微泡+超声照射。制作兔急性心梗模型,分别用上述三种方法从静脉导入hAng-1基因。2周后心肌造影检测梗死心肌灌注状况聚合酶链反应(RT-PCR)检测hAng-1基因的mRNA表达以评价转染疗效。结果携抗ICAM-1抗体的微泡在体外能大量靶向结合到产生炎性反应的ECV304细胞周边。应用靶向微泡转染2周后,左室内径减小,射血分数升高,但与非靶向微泡比较,差异无统计学意义(P>0.05),而心肌造影显示心肌内造影剂填充量较非靶向微泡组增多,且RT-PCR定量灰度分析hAng-1mRNA(1.04±0.08)高于非靶向微泡组(0.53±0.04),差异有统计学意义(P<0.01)。结论微泡与抗ICAM-1抗体连接后形成靶向微泡载体,可明显提高hAng-1基因在体内的转染效率,增加心肌梗死后局部的血管新生效应。

Objective To investigate whether the anti - ICAM - 1 targeted microbubble can improve the transfection efficiency of hAng - 1 gene in myocardial infarction. Methods The combination degree of anti - ICAM - 1 targeted microbubbles with human interleukin - 1 βstimulated ECV304 cells was detected in vitro. And for animal study, myocardial infarction rabbits were divided into three groups： control group, non -targeted microbubbles delivery group and targeted microbubble delivery group. The hAng - 1 gene was transferred into vein by three methods under ultrasound irradiation. The condition of infarcted myocardial perfusion was detected by cardiomyography after two weeks, and mRNA expression of hAng - 1 gene was detected by RT - PCR, which was used to assess gene transfection efficacy. Results In vitro, a large number of anti - ICAM - 1 microbubbles could targeted combine with ECV304 ceils which responsed to inflammatory reaction. After two weeks of gene transfection, the diameter of left ventricular was decreased, and LVEF was increased, while there was no significant difference between targeted microbubble delivery group and non - targeted microbubbles delivery group （ P 〈 0. 05 ） . And cardiomyography showed that contrast agent filling in myocardium was more than that in non - targeted microbubbles delivery group, and the hAng - 1 mRNA by RT - PCR quantitative analysis was higher than that in non - targeted microbubbles delivery group（1.04±0.08 vs0.53 ± 0. 04 , P 〈 0. 01） . Conclusion Microbubblecarryinganti-ICAM-1 andhAng-1 geneformeda new type of targeted microbubble, it can significantly improve the transfeetion efficacy of hAng - 1 gene expression in vivo, and it also can improve the partial angiogenesis effects after myocardial infarction.