Antisense MMP-9 oligonucleotide suppresses aneurysmal formation in vivo

本实验旨在探讨基质金属蛋白酶9（MMP-9）反义寡核苷酸对肿瘤形成的抑制作用。0.5在100 mmHg下将1 ml（25单位/ml）的胰弹性蛋白酶输注到大鼠肾下腹主动脉中30分钟。弹性蛋白酶灌注后主动脉平均直径显著增加（灌注前1.276±0.119 mm，灌注后2.143±0.145 mm，P<0.0001）。将浸渍有反义MMP-9寡核苷酸的Pluronic凝胶（n=8）应用于注入弹性蛋白酶的主动脉的外部。将具有随机寡核苷酸的Pluronic凝胶（n=8）或单独的Pluronic凝胶（n=7）也应用于主动脉作为对照。术后7天处死大鼠，测量主动脉直径。将部分酵母菌置于液氮中进行酶谱分析和蛋白质印迹分析。用反义MMP-9寡核苷酸处理的主动脉的直径在第7天时显著小于接受随机寡核苷酸或单独载体的主动脉的直径（反义寡核苷酸; 5.362±1.948 mm，随机寡核苷酸; 7.852±2.789 mm，普朗尼克凝胶; 7.524±2.074 mm，p<0.05）。反义寡核苷酸组的直径增加率也显著低于随机寡核苷酸组和赋形剂组（反义寡核苷酸组：2.510±0.956;随机寡核苷酸组：3.746±1.268;普朗尼克凝胶组：3.503±1.115，P<0.05）。通过使用酶谱法和蛋白质印迹法，我们现在进行MMP-9蛋白检测，以调查是否反义MMP-9寡核苷酸抑制MMP-9活性在主动脉和培养的大鼠腹腔巨噬细胞。结论：MMP-9反义寡核苷酸有望成为抑制主动脉瘤的一种可行的治疗方法。

The purpose of this experimental study was to explore the inhibitory effect of antisense matrix metallorpoteinase-9 (MMP-9) oligonucleotide on the suppression of aneurysmal formation in vivo. 0.5 ml (25 units/ml) of pancreatic elastase was infused into rat infrarenal abdominal aorta at 100 mmHg for 30 minutes. Mean diameter of the aorta was significantly increased after the elastase infusion (before; 1.276±0.119 mm, after; 2.143±0.145 mm, P<0.0001). Pluronic gel impregnated with antisense MMP-9 oligonucleotide (n=8) was applied on the exterior of elastase-infused aorta. Pluronic jel with randomized oligonucleotide (n=8) or pluronic gel alone (n=7) was also applied on the aorta as control. Rats were sacrificed 7 days after surgery, and diameter of the aorta was measured. Some aortas were put into liquid nitrogen for zymography and western blot analysis. Diameter of the aorta treated with antisense MMP-9 oligonucleotide was significantly less at 7 days compared with that received the randomized oligonucleotide or vehicle alone (antisense oligo; 5.362±1.948 mm, randomized oligo; 7.852±2.789 mm, pluronic jel; 7.524±2.074 mm, p<0.05). The increased ratio of diameter was also significantly less in the antisense oligonucleotide group than in the randomized oligonucleotide or vehicle (antisense oligo; 2.510±0.956, randomized oligo; 3.746±1.268, pluronic jel; 3.503±1.115, P<0.05). By using zymography and western blotting, we are now conducting MMP-9 protein assays for investigating whether antisense MMP-9 oligonucleotide inhibits MMP-9 activity in aorta and in cultured rat peritoneal macrophages. In conclusion, antisense MMP-9 oligonucleotide may promise a feasible treatment for inhibiting aortic aneurysm.