Application of real-time RT-PCR to quantifying gene expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in human abdominal aortic aneurysm
Application of real-time RT-PCR to quantifying gene expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in human abdominal aortic aneurysm
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DOI：
10.1016/j.atherosclerosis.2004.07.013
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发表时间：
2004-12-01
期刊：
影响因子：
5.3
通讯作者：
Sakamoto, A
中科院分区：
文献类型：
--
作者：
Higashikata, T;Yamagishi, M;Sakamoto, A
背景：基质金属蛋白酶（MMPs）和金属蛋白酶组织抑制剂（TIMPs）是细胞外基质重塑的关键调节因子，其相对表达水平在腹主动脉瘤（AAA）的发生发展中起着关键作用。然而，很少有数据存在定量评估其在临床环境中的表达。方法：22例接受人工血管置换术的AAA患者，获得AAA和未扩张主动脉的组织标本。使用实时RT-PCR方法，使小组织样本中的mRNA水平的定量测量，我们确定了MMPs和TIMPs的基因表达水平相对于戊二醛3-磷酸脱氢酶在每个样品中。结果如下：MMP-1和MMP-3基因的表达水平在AAA中显著高于非扩张区（分别为4.48 ± 2.01和0.26 ± 0.12，P < 0.01; 1.89 ± 1.00和5.01 ± 0.97，P < 0.05）。尽管TIMP-1、TIMP-2和TIMP-3的基因在AAA中有上调的趋势，但MMP-1与TIMP-1、MMP-1与TIMP-2、MMP-1与TIMP-3和MMP-3与TIMP-2的相对表达水平在AAA中仍高于非扩张区（分别为1.12 +/- 0.63 vs 0.10 +/- 0.03，4.13 +/- 1.12 vs 0.43 +/- 0.11，1.61 +/- 0.59 vs 0.14 +/- 0.03，7.81 +/- 1.60 vs 2.56 +/- 0.76，P < 0.05）。结论：这些结果表明，目前的实时RT-PCR方法是可靠的，用于测定血管组织的小样本中的mRNA水平，MMP-1和MMP-3相对于TIMP的不成比例的表达与AAA的病理演变有关。(C)2004爱思唯尔爱尔兰有限公司保留所有权利。
Backgound: The relative expression levels of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs), key regulators in remodeling of extracellular matrix, are considered to play a pivotal role in the development of abdominal aortic aneurysm (AAA). However, few data exist regarding quantitative assessment of their expression in clinical settings. Methods: In 22 patients with AAA who underwent graft replacement, tissue samples of the AAA and non-dilated aorta were obtained. Using a real-time RT-PCR method that enabled quantitative measurement of mRNA levels in small tissue samples, we determined gene expression levels of MMPs and TIMPs relative to that of glutaraldehyde 3-phosphate dehydrogenase in each sample. Results: The expression levels of the MMP-1 and -3 genes were significantly augmented in AAA compared with non-dilated regions (4.48 +/- 2.01 versus 0.26 +/- 0.12, P < 0.01 and 1.89 +/- 1.00 versus 5.01 +/- 0.97, P < 0.05, respectively). Although genes for TIMP-1, -2 and -3 tended to be upregulated in AAA, relative expression levels of MMP-1 to TIMP-1, MMP-1 to TIMP-2, MMP-1 to TIMP-3, and MMP-3 to TIMP-2 were still higher in AAA than in non-dilated regions (1.12 +/- 0.63 versus 0.10 +/- 0.03, 4.13 +/- 1.12 versus 0.43 +/- 0.11, 1.61 +/- 0.59 versus 0.14 +/- 0.03, and 7.81 +/- 1.60 versus 2.56 +/- 0.76, respectively, P < 0.05). Conclusion: These results demonstrate that the present real-time RT-PCR method is reliable for the determination of mRNA levels in small samples of vascular tissue and that disproportional expression of both MMP-1 and MMP-3 relative to TIMPs relates pathologically to the evolution of AAA. (C) 2004 Elsevier Ireland Ltd. All rights reserved.