摘要
目的探讨注射加压转染的方法对兔动脉壁进行人凝血酶调节蛋白(human thrombomodulin,hTM)基因转染的可行性。方法健康新西兰大白兔84只,应用抽签法随机分为3组:pcDNA3.1/hTM质粒组(n=28),pcDNA3.1(+)/neo质粒组(n=28)和未转染组(n=28),通过基因转染并建立动脉损伤-阻滞模型,于术后3、7、14和28 d分别检测hTM mRNA和蛋白在动脉壁的表达。结果17只实验动物在手术当天至术后3 d内意外死亡。pcDNA3.1/hTM质粒组各时间点的hTM mRNA表达都明显高于pcDNA3.1(+)/neo质粒组和未转染组(P<0.01);pcDNA3.1(+)/neo质粒组和未转染组组内和组间每个时间点的hTM mRNA表达比较,差异均无统计学意义(P>0.05)。hTM蛋白在各组中均有表达,主要位于动脉内腔面;pcDNA3.1/hTM质粒组各时间点的hTM蛋白表达明显高于pcDNA3.1(+)/neo质粒组和未转染组(P<0.05);pcDNA3.1(+)/neo质粒组和未转染组的hTM蛋白表达在各个时间点都相对恒定,组内和组间比较差异均无统计学意义(P>0.05)。结论用动脉腔内加压法可以有效地对活体动物的动脉壁进行基因转染。
Objective To explore the feasibility of high-pressure injection to transfer human thrombomodulin （hTM） gene into arterial wall of rabbits. Methods Eighty-four healthy New Zealand rabbits were randomly divided into three groups： pcDNA3.1/hTM plasmid group （n=28）, peDNA3, 1（＋）/neo plasmid group （n=28） and untransfected group （n= 28）. After gene transfection, the model of arterial injury-blocking was established. Then, the expressions of hTM mRNA and protein in arterial wall were examined by RT-PCR and immunohistochemistry at 3 d, 7 d, 14 d and 28 d after operation. Results Seventeen rabbits died accidentally from the day of operation to 3 d after operation. The expressions of hTM mRNA of different time points in pcDNA3. 1/hTM plasmid group were significantly higher than that in pcDNA3, 1（＋）/neo plasmid group and untransfected group （P〈0.01）. For the expressions of hTM mRNA at different time points in pcDNA3.1 （＋）/neo plasmid group and untransfeeted group, the difference of inter-group and intra-group was not significant （P〉0.05）. hTM protein was expressed in every group and mainly localized in the inner lining of arterial wall. The expressions of hTM protein at different time points in pcDNA3.1/hTM plasmid group were significantly higher than that in pcDNA3. 1 （＋）/neo plasmid group and untransfeeted group （P〈0.05）. The expression of hTM protein at different time points in pcDNA3.1（＋）/neo plasmid group and untransfected group kept relative constancy, the difference of inter-group and intra-group was also not significant （P〉0.05）. Conclusion High-pressure injection is feasible to transfer pcDNA3.1/hTM plasmid into arterial wall of live animals.
出处
《中国普外基础与临床杂志》
CAS
2009年第8期628-632,共5页
Chinese Journal of Bases and Clinics In General Surgery
关键词
人凝血酶调节蛋白
基因转染
动脉
兔
Human thrombomodulin
Gene transfection
Artery
Rabbit
