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Antisense MMP-9 oligonucleotide suppresses aneurysmal formation in vivo

反义 MMP-9 寡核苷酸抑制体内动脉瘤形成

基本信息

批准号：
12671160
负责人：
ZEMPO Nobuya
金额：
$ 2.18万
依托单位：
Yamaguchi University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

项目摘要

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.

本实验旨在探讨反义基质金属蛋白酶-9 （MMP-9）寡核苷酸在体内对动脉瘤形成的抑制作用。胰弹性蛋白酶0.5 ml（25单位/ml）以100 mmHg滴入大鼠肾下腹主动脉30分钟。注射弹性蛋白酶后主动脉平均直径明显增加（注射前1.276±0.119 mm，注射后2.143±0.145 mm， P<0.0001）。将含有反义MMP-9寡核苷酸的Pluronic凝胶（n=8）涂于注入弹性酶的主动脉外壁。随机寡核苷酸Pluronic凝胶（n=8）或单独Pluronic凝胶（n=7）也应用于主动脉作为对照。术后7 d处死大鼠，测量主动脉直径。将部分主动脉置于液氮中进行酶谱分析和western blot分析。7 d时，反义MMP-9寡核苷酸组主动脉直径明显小于单纯随机寡核苷酸组或单纯载体组（反义寡核苷酸组；5.362±1.948 mm，随机寡核苷酸组；7.852±2.789 mm， pluronic凝胶组；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或载体组（反义寡核苷酸组；随机寡核苷酸组；3.746±1.268组；3.503±1.115组，P<0.05）。通过酶谱法和western blotting，我们正在进行MMP-9蛋白检测，以研究反义MMP-9寡核苷酸是否抑制主动脉和培养的大鼠腹腔巨噬细胞中MMP-9的活性。总之，反义MMP-9寡核苷酸可能是抑制主动脉瘤的一种可行的治疗方法。