Castier Y, Chemla E, Nierat J, Heudes D, Vasseur M A, Rajnoch C, Bruneval P, Carpentier A, Fabiani J N
Department of Cardiovascular Surgery, Broussais Hospital, Paris, France.
J Cardiovasc Surg (Torino). 1998 Feb;39(1):1-7.
We sought to determine the efficacy and specificity of a new c-myb antisense by inhibiting neointimal hyperplasia in a rat abdominal aorta injury model. Using c-myb antisense oligonucleotides, inhibition of vascular smooth muscle cell proliferation has been reported.
Sixty-six male Wistar rats had a de-endothelialization of the abdominal aorta. Following a double blind randomization protocol, F127 pluronic gel containing one of the five oligonucleotides or plain gel was applied around the aorta: 1) 18-mer c-myb antisense (AS18) with four contiguous guanosines (G-quartet); 2) 15-mer c-myb antisense (AS15) without G-quartet; 3) 1-bp mismatch AS15 without G-quartet (MM1); 4) an oligonucleotide with G-quartet (4G), whereas the other bases were chosen at random; 5) 1-bp mismatch 4G without G-quartet (MM2). After 21 days all rats were sacrificed and aortas harvested for histomorphometric evaluation. Four rats were given fluorescent-labeled oligonucleotides to study in vivo localization after local advential delivery.
Morphometric analysis showed significant suppression of neointimal hyperplasia in AS18 and 4G and MM2 groups compared with GEL, AS15 and MM1 groups (p<0.05). The oligonucleotide-labeled aortas showed penetration of the oligonucleotides into the media which increased with time.
Our findings pointed to the potential non specificity of the c-myb antisense oligonucleotide in vivo. Such results will minimize the importance of antisense strategy as a potential therapeutic for preventing neointimal hyperplasia. The two oligonucleotides with a G-quartet inhibited neointimal hyperplasia in our model. Exploring a non-antisense mechanism, G-quartet oligonucleotides as potential drugs to reduce neointimal hyperplasia is attractive.
我们试图通过抑制大鼠腹主动脉损伤模型中的内膜增生来确定一种新的c-myb反义核酸的疗效和特异性。已有报道称，使用c-myb反义寡核苷酸可抑制血管平滑肌细胞增殖。
66只雄性Wistar大鼠接受腹主动脉去内皮处理。按照双盲随机方案，将含有五种寡核苷酸之一的F127普朗尼克凝胶或普通凝胶应用于主动脉周围：1）含有四个连续鸟苷（G-四联体）的18聚体c-myb反义核酸（AS18）；2）不含G-四联体的15聚体c-myb反义核酸（AS15）；3）不含G-四联体的1个碱基错配的AS15（MM1）；4）含有G-四联体的寡核苷酸（4G），而其他碱基随机选择；5）不含G-四联体的1个碱基错配的4G（MM2）。21天后，处死所有大鼠并采集主动脉进行组织形态计量学评估。给4只大鼠注射荧光标记的寡核苷酸，以研究局部外膜给药后的体内定位。
形态计量学分析显示，与凝胶组、AS15组和MM1组相比，AS18组、4G组和MM2组的内膜增生受到显著抑制（p<0.05）。寡核苷酸标记的主动脉显示寡核苷酸渗透到中膜，且随时间增加。
我们的研究结果表明c-myb反义寡核苷酸在体内可能存在非特异性。这些结果将降低反义策略作为预防内膜增生潜在治疗方法的重要性。在我们的模型中，两种含有G-四联体的寡核苷酸抑制了内膜增生。探索一种非反义机制，将G-四联体寡核苷酸作为减少内膜增生的潜在药物具有吸引力。
