联合转染反义PDGF寡核苷酸和tPA基因抑制血管内膜增生的实验研究  

Effects of platelet derived growth factor antisense oligodeoxynucleotides and tissue-type plasminogen activator gene transfection on inhibition of intimal proliferation

作　　者：吴忠均[1] 时德[2] 李德卫[2] 郑树森[1] 

机构地区：[1]浙江大学医学院附属第一医院普通外科,浙江杭州310003 [2]重庆医科大学附属第一医院血管外科,重庆400016

出　　处：《浙江大学学报（医学版）》2005年第4期320-325,338,共7页Journal of Zhejiang University(Medical Sciences)

基　　金：国家重点基础研究发展规划(973)基金资助项目(2003CB515501);国家自然科学基金资助项目(30270514)

摘　　要：目的:观察血管局部联合转染血小板源性生长因子(PDGF)反义寡核苷酸(AODN)和组织型纤溶酶原激活物(tPA)基因对自体移植动脉内膜增生的影响,并探讨其可能机制。方法:选100只新西兰雄性白兔,随机分为4组(每组25只):A组(对照组)、B组(PDGF-AODN转染组)、C组(tPA基因转染组)、D组(PDGF-AODN和tPA基因联合转染组)。将同一只兔的左、右髂外动脉(各1.0cm)对换移植,移植血管分别用PDGF-AODN和pBudCE4.1/tPA液浸泡,血管吻合口用上述液体浸泡过的缝线吻合。每组按实验终点(术后3d、1w、2w、4w和8w)分为5个亚组,术后各实验终点取移植血管标本,用于病理学检测、电镜观察、发色底物检测tPA活性实验3、H-TdR掺入实验和免疫组织化学染色检测。结果:术后各时间点D组移植血管的内膜面积、管腔狭窄程度、3H-TdR掺入量和PDGF阳性细胞数均显著低于A组(P<0.01),而且亦明显低于B和C组(P<0.05)。扫描电镜观察显示C组和D组血管壁只见少量血小板附着,未见血栓形成,而A组和B组血管壁可见大量血小板附着,且有血栓形成。术后3d、1w、2w、4w时C组与D组的tPA基因表达产物活性明显高于A组(P<0.01)。结论:血管局部联合转染PDGF-AODN和tPA基因能有效抑制VSMC增殖,阻止内膜增生,防止移植血管狭窄。Objective： To observe the effects of co-transfection of platelet derived growth factor antisense oligodeoxynucleotides （PDGF-AODN） and tissue-type plasminogen activator （tPA） gene on inhibition of intimal proliferation of auto-transplantion artery, Methods ： One hundred male New Zealand rabbits were randomly divided into four groups （25 in each）,,Group A （control group）,Group B （PDC-F-AODN transfection group）,Group C （tPA gene transfection group） and Group D（PDGF-AODN and tPA co-transfection group）. The left and right external iliac arteries were transplanted reciprocally. The transplanted arteries were respectively soaked in PDGFAODN, pBudCE4. 1/tPA and PDGF-AODN plus pBudCE4. 1/tPA solution about 15 minute before transplantation, The rabbits were sacrificed at 3d, 1w, 2w, 4w and 8w after operation. The specimens were harvested for pathologic examination,electron microscopy,chromogenic substrate test,^3H-TdR incorporation test and immunohistochemical staining, Results： The scan electron microscopy showed that there were a few thrombocytes on vas-wall of Group C and D without thrombus,whereas there were abundant thrombocytes and thrombus forming on vas-wall of Group A and B, The intimal area,stenosis ratio of transplanted artery,^3H-TdR incorporation,the number of PDGF positive cell in Group D were significantly less than those in Group A （P〈0.01） ,Group B and Group C （both P〈0.05）. The activity of tPA gene products in transplanted vas-wall of Group D was significantly higher than that of Group A （P〈0.01）. Conclusion.. Local co-transfection of PDGF-AODN and tPA gene can effectively inhibit the proliferation of vascular smooth muscle cells,hyperplasia of intima and restenosis of transplanted artery.

关 键 词：寡核苷酸类 反义/分析 血小板源生长因子/分析 组织型纤溶酶原激活物 基因表达 

分 类 号：R543[医药卫生—心血管疾病]