转染Angiopoietin-1基因对急性心梗后左室功能的影响  

The Effect of Transferring Angiopoietin-1 Gene Into the Left Ventricular After Acute Myocardial Infarction In Rabbit

作　　者：陈仕林[1] 刘玉清[1] 朱成楚[2] 叶加洪[2] 叶中瑞[2] 郭剑波[2] 叶敏华[2] 马德华[2] 

机构地区：[1]中国医学科学院中国协和医科大学阜外心血管病医院中德分子医学实验室,北京市100037 [2]浙江省台州医院胸心外科,317000

出　　处：《中国分子心脏病学杂志》2005年第2期457-462,F0003,共7页Molecular Cardiology of China

基　　金：国家人事部博士后基金资助项目(编号:2002032188)

摘　　要：目的探讨重组腺病毒载体介导的血管形成素1(Ang1)基因转染对兔急性心梗后左室功能影响。方法构建含有Ang1的腺病毒粘粒载体,通过转染293细胞进行同源重组制备重组腺病毒颗粒。64只雄性新西兰大白兔行高位冠状动脉结扎,4只用于急性心梗后循环中VEGF含量的检测,其余60只随机均分为实验组、单纯培养基组(DMEM)与LacZ重组腺病毒组,分别于冠状动脉结扎后心肌内直接注射Ang1重组腺病毒,DMEM与LacZ重组腺病毒。并于转染后第3、14、28d应用超声心动图观察心功能情况,第3、7、14、28d应用RT PCR方法测定重组腺病毒基因在转染心肌中的表达,第14、28d用免疫组化方法观察缺血心肌内血管生长情况。结果获得的Ang1重组腺病毒滴度为5.6×1011pfu/L。重组腺病毒直接注射转染兔缺血心肌后能有效表达目的基因,RT PCR测定提示Ang1基因转染后第3d心肌中即有表达,7、14d仍呈阳性表达,28d未测出。心肌梗死后Ang1组心功能从术后第3d至第28d得到明显改善,其幅度显著好于对照组。心肌梗死后左室心肌收缩期截面积与舒张期截面积均明显增加,尤以术后第3d为著,随时间延长均有所缩小,至28d后Ang1组缩小幅度明显高于其他两组。转染14、28d后Ang1组新生毛细血管密度及αSMA阳性的小动脉性血管密度均明显高于对照组(P<0.01)。结论腺病毒介导的Ang1基因在兔缺血心肌中能有效地促进新生血管形成,并能改善缺血心肌的功能。Objective To explore the effect of human angiopoietin-1 （Ang1） gene mediated by recombinant adenovirus on the left ventricular function changes in ischemic rabbit myocardium . Methods A adenovirus cosmid vector containing human Angl gene was engineered and transferred into 293 host cells by homolegous recombination to produce adenovirus. Sikty-fourth male New Zealand white rabbits underwent high positioned double-ligation of the anterior descending left coronary artery. Four rabbits were used to determine the level of VEGF in the circulation of acute myocardial infarction , and the rest divided into three groups： Ang1 group （n = 20） received direct myocardium injection of angiopoietin-1 recombinant adenovims; DMEM group （n = 20） and LacZ group （n = 20） received only DMEM and LacZ recombinant adenovims as control respectively. The left ventricular function was evaluated by echocardiography on the 3^th, 14^th, 28^th postoperative day and the expression of Angl was determined by RT-PCR on the 3^th ,7^th, 14^th ,28^th postoperative day . The degree of angiogenesis in the ischemic myocardium was calculated on the 14^th ,28^th postoperative day by use of immunohistochemical analysis . Results The results showed that the replication-deficient recombinant adenovirus of Angl was generated by homologous recombination, with the titers of 5.6 × 10^11 pfu/L. In our rabbit acute myocardial infarction model, serum VEGF level increased within 12h and peaked at 24h after left anterior descending coronary artery ligation. This high concentration of VEGF lasted for 72 h. Animals treated with Adenoviral Angl demonstrated a significant increase in all measures of left ventricular function between day 3 and 28 post-infarction, these improvements were obviously greater than those in control animals. The results from RT-PCR showed that the Angl gene could be transcripted in vivo on 3th jth ,14th postoperative day but not on 28^th postoperative day and control groups. Immunohistochemical analysis revealed that the capi

关 键 词：血管形成素-1 心肌梗死 基因治疗 心功能 

分 类 号：R542.22[医药卫生—心血管疾病]