Altered expression balance of matrix metalloproteinases and their inhibitors in human carotid plaque disruption: Results of quantitative tissue analysis using real-time RT-PCR method
Altered expression balance of matrix metalloproteinases and their inhibitors in human carotid plaque disruption: Results of quantitative tissue analysis using real-time RT-PCR method
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DOI：
10.1016/j.atherosclerosis.2005.05.039
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发表时间：
2006-03-01
期刊：
影响因子：
5.3
通讯作者：
Akamoto, A
中科院分区：
文献类型：
--
作者：
Higashikata, T;Yamagishi, M;Akamoto, A
背景：动脉粥样硬化组织中细胞外基质的降解与合成的平衡决定了其含量。为探讨基质金属蛋白酶（MMPs）及其抑制剂、金属蛋白酶组织抑制剂（TIMPs）和组织因子途径抑制剂-2（TFPI-2）表达平衡在动脉粥样硬化斑块形成和破坏中的作用，采用实时荧光定量逆转录-聚合酶链反应（RT-PCR）方法检测了人颈动脉斑块中MMPs及其抑制剂、TIMPs和TFPI-2的表达。从24例颈动脉内膜切除术患者的组织中提取用于cDNA合成的总RNA。结果：在斑块中，MMP-1（1.53 ± 0.25，mean +/-S.E.M.），TIMP-1（1.53 ± 0.25，mean +/-S.E.M.），TIMP-2（1.53 ± 0.25，mean +/-S.E.M.），MMP-3（1.53 ± 0.25，mean +/-S.E.M.），TIMP-1（1.53 ± 0.25，mean +/-S.E.M.），TIMP-3（1.53 ± 0.25，mean +/-S.E.M.），MMP-3（1.53 ± 0.25，mean +/-S.E.M.），TIMP-1（1.53 ± 0.25，mean +/-S.E.M.），TIMP-1（1.53 ± 0.25，mean +/-S.E.M.），MMP MMP-3（1.99 +/-0.59）和MMP-9（2.00 +/-0.51）与相邻对照区相比增加（分别为0.60 +/-0.16、0.46 +/-0.18和0.58 +/-0.21，p <0.05）。斑块中TFPI-2的表达（0.32 ± 0.08）低于对照组（0.94 ± 0.23，p <0.01）。尽管斑块中TIMP-1的表达（1.28 ± 0.23）高于对照组（0.81 ± 0.10，p <0.05），但斑块中MMP-1/TIMP-1、MMP-3/TIMP-3和MMP-9/TIMP-1的指数仍显著高于对照组。有趣的是，MMP-9和由此产生的MMP-9/TIMP-1平衡在斑块破裂中显著升高，（3.36 +/-1.52和1.66 +/-0.12，n = 11）（1.11 +/-0.52和0.76 +/-0.12，n = 13，p <0.05）。随着TFPI-2表达的降低，动脉粥样硬化斑块中MMPs的上调与TIMPs的上调不成比例，这表明在晚期病变中，特别是在破裂的斑块中，细胞外基质的不平衡降解和合成持续存在。(c)2005爱思唯尔爱尔兰有限公司保留所有权利。
Background: The balance between degradation and synthesis of extracellular matrix determines its content in atherosclerotic tissue. To examine the role of expression balance of matrix metalloproteinases (MMPs) to their inhibitors, tissue inhibitors of metalloproteina ses (TIMPs) and tissue factor pathway inhibitor-2 (TFPI-2) in the development and disruption of atherosclerotic plaque, these gene expressions in human carotid plaque were quantitatively determined by real-time reverse transcription (RT)-polymerase chain reaction (PCR) method.Methods: Total RNA for cDNA synthesis was extracted from tissues in 24 patients with carotid endarterectomy. The amounts of cDNAs for MMP-1, -2, -3 and -9, TFPI-2 and TIMP-1, -2 and -3 were determined by real-time RT-PCR method, and normalized with glutaraldehyde 3-dehydrogenase.Results: In plaques, the expression MMP-1 (1.53 +/- 0.25, mean +/- S.E.M.), MMP-3 (1.99 +/- 0.59) and MMP-9 (2.00 +/- 0.51) was augmented compared to those in the adjacent control regions (0.60 +/- 0.16, 0.46 +/- 0.18 and 0.58 +/- 0.21, respectively, p < 0.05). The expression of TFPI-2 was lower in plaques (0.32 +/- 0.08) than in controls (0.94 +/- 0.23, p < 0.01). Although the expression of TIMP-1 was higher in plaques (1.28 +/- 0.23) than in controls (0.81 +/- 0.10, p < 0.05), the indices of MMP-1/TIMP-1, MMP-3/TIMP-3 and MMP-9/TIMP-1 were still significantly higher in plaques. Interestingly, MMP-9 and the resulting MMP-9/TIMP-1 balance in plaques with disruption were significantly higher (3.36 +/- 1.52 and 1.66 +/- 0.12, n = 11) than those in non-disrupted plaques (1.11 +/- 0.52 and 0.76 +/- 0.12, n = 13, p < 0.05).Conclusion: With the decreased expression of TFPI-2, upregulation of MMPs in atherosclerotic plaque was disproportional to that of TIMPs, suggesting that imbalanced degradation and synthesis of extracellular matrix persists in advanced lesions, particularly in plaques with disruption. (c) 2005 Elsevier Ireland Ltd. All rights reserved.