Preoperative treatment with doxycycline reduces aortic wall expression and activation of matrix metalloproteinases in patients with abdominal aortic aneurysms
Preoperative treatment with doxycycline reduces aortic wall expression and activation of matrix metalloproteinases in patients with abdominal aortic aneurysms
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DOI：
10.1016/s0741-5214(00)90163-0
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发表时间：
2000-02-01
影响因子：
4.3
通讯作者：
Thompson, RW
中科院分区：
文献类型：
--
作者：
Curci, JA;Mao, DL;Thompson, RW
目的：基质金属蛋白酶（MMPs）被认为在腹主动脉瘤（AAA）的发病机制中起核心作用。多西环素（Dox）在体外具有直接的MMP抑制特性，并且它有效地抑制啮齿动物中弹性蛋白酶诱导的AAA的发展。本研究的目的是确定是否与Dox治疗抑制基质金属蛋白酶在人类动脉瘤tissue.Methods：动脉瘤组织的分子机制，阐明这种影响的基础上，从15例AAA，其中8人已与Dox治疗手术前（100毫克口服7天，每天两次）。通过明胶酶谱和免疫印迹分析检测蛋白质提取物，通过逆转录-聚合酶链反应（RT-PCR）检测RNA。结果：用底物酶谱法测定MMP总活性，两组间无明显差异，但Dox处理组MMP活性与对照组相比有轻微差异，而Dox处理组MMP活性与对照组相比无显著差异。72-kDa明胶酶的活化部分减少（24.4%）（MMP-2; P <0.05），相反，免疫印迹分析显示，在Dox治疗的患者中，可提取的92-kDa明胶酶（MMP-9）蛋白的量减少了2.5倍（P <0.05）。此外，通过定量竞争性RT-PCR证实了Dox治疗的患者中MMP-9信使RNA（mRNA）减少5.5倍（81.9%）（平均值+/- SE，mol MMP-9/mol β-肌动蛋白：1.3 +/- 0.5 vs 7.2 +/- 3.1; p <0.04）。各组间MMP-2蛋白或mRNA的相对表达无显著差异。体外培养的THP-1单核细胞经佛波酯刺激后，MMP-9蛋白和mRNA的表达均受到相应浓度Dox的抑制。除了其作为直接MMP拮抗剂的公认作用外，Dox可能通过降低单核细胞/巨噬细胞MMP-9 mRNA的表达和抑制翻译后加工来影响人类动脉瘤组织内的结缔组织降解。（活化）proMMP-2。通过这种机制的互补组合，Dox治疗可能是AAA患者实现MMP抑制的特别有效的策略。
Purpose: Matrix metalloproteinases (MMPs) are considered to play a central role in the pathogenesis of abdominal aortic aneurysms (AAAs). Doxycycline (Dox) has direct MMP-inhibiting properties in vitro, and it effectively suppresses the development of elastase-induced AAAs in rodents. The purpose of this study was to determine if treatment with Dox suppresses MMPs within human aneurysm tissue and to elucidate the molecular mechanisms underlying this effect.Methods: Aneurysm tissues were obtained from 15 patients with an AAA, eight of whom had been treated with Dox before surgery (100 mg orally twice a day for 7 days). Protein extracts were examined by means of gelatin zymography and immunoblot analysis, and RNA was examined by means of reverse transcription-polymerase chain reaction (RT-PCR). The effects of Dox on MMP production were further examined in human THP-1 mononuclear phagocytes in vitro.Results: NO detectable difference was found between groups by using substrate zymography as a means of assessing total MMP activity, but Dox treatment was associated with a slight (24.4%) reduction in the activated fraction of 72-kDa gelatinase (MMP-2; P < .05), In contrast, a 2.5-fold reduction in the amount of extractable 92-kDa gelatinase (MMP-9) protein in Dox-treated patients was revealed by means of immunoblot analysis (P < .05). Also, a 5.5-fold (81.9%) reduction in MMP-9 messenger RNA (mRNA) in Dox-treated patients was demonstrated by means of quantitative competitive RT-PCR (mean +/- SE, mol MMP-9/mol beta-actin: 1.3 +/- 0.5 vs 7.2 +/- 3.1; p < .04). There was no significant difference between groups in the relative expression of MMP-2 protein or mRNA. In cultured THP-1 monocytes stimulated with phorbol ester, the expression of MMP-9 protein and mRNA were both decreased after exposure to relevant concentrations of Dox in vitro.Conclusion: In addition to its recognized effects as a direct MMP antagonist, Dox may influence connective tissue degradation within human aneurysm tissue by reducing monocyte/macrophage expression of MMP-9 mRNA and by suppressing the post-translational processing (activation) of proMMP-2. Through this complementary combination of mechanisms, treatment with Dox may be a particularly effective strategy for achieving MMP inhibition in patients with an AAA.