Matrix metalloproteinases and tissue inhibitors of metalloproteinases in coxsackievirus-induced myocarditis
Matrix metalloproteinases and tissue inhibitors of metalloproteinases in coxsackievirus-induced myocarditis
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DOI：
10.1016/j.carpath.2005.11.008
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发表时间：
2006-03-01
影响因子：
3.7
通讯作者：
McManus, BM
中科院分区：
文献类型：
--
作者：
Cheung, C;Luo, HL;McManus, BM
背景：柯萨奇病毒B3 （CV133）是人类心肌炎的主要病原体。在小鼠模型中，心肌炎的炎症期导致心脏的广泛损伤，并引发细胞外基质（ECM）的深度重塑，最终可能导致扩张型心肌病。基质金属蛋白酶（MMPs）是ECM的调节因子，可以降解基质中的所有成分。方法：用心毒性CV133感染青春期雄性小鼠，分别于感染后3、9、30天处死。采用逆转录聚合酶链反应（RT-PCR）检测MMP-2、MMP-9和MMP-12的转录水平。免疫组织化学检测这些酶的蛋白表达，明胶酶谱法检测MMP-2和MMP-9的激活状态。采用免疫印迹法分析金属蛋白酶组织抑制剂（TIMPs）。心肌用小天狼星红染色，在偏振光下观察胶原网络。结果：通过RT-PCR检测，MMP-2、MMP-9和MMP-12的转录在每隔9天增加。免疫组织化学证实这些MMP物种的翻译增加，酶谱分析进一步显示CV133感染后MMP-2和MMP-9的激活升高。TIMP-3和TIMP-4的表达下调，而TIMP-1和TIMP-2在整个感染过程中保持不变。用小天狼星红染色的小鼠心脏显示，在感染的急性期，胶原蛋白总量增加，在随后的时间点，原纤维被破坏。结论：CV133感染后，ECM重构被触发，这种反应可能与MMPs的表达和激活增加有关。(C) 2006爱思唯尔公司版权所有。
Background: Coxsackievirus B3 (CV133) is the major causative agent of myocarditis in humans. In the mouse model, the inflammatory phase of myocarditis results in extensive damage to the heart and triggers profound extracellular matrix (ECM) remodeling, which may ultimately lead to dilated cardiomyopathy. Matrix metalloproteinases (MMPs) are regulators of the ECM and can degrade all the components in the matrix. Methods: Adolescent male mice were infected with cardiovirulent CV133 and sacrificed at 3, 9, and 30 days post infection (pi). Transcription of MMP-2, MMP-9, and MMP-12 was assessed by reverse-transcriptase polymerase chain reaction (RT-PCR). Protein expression of these enzymes was examined using immunohistochemistry, and the activation status of MMP-2 and MMP-9 was assessed using gelatin zymography. Tissue inhibitors of metalloproteinases (TIMPs) were analyzed using immunoblotting assays. Myocarditic hearts were also stained with picrosirius red and viewed under polarizing light to examine the collagen network. Results: MMP-2, MMP-9, and MMP-12 transcription was increased at 9 days pi, as determined by RT-PCR. Immunohistochemistry confirmed an increase in translation of these MMP species, and zymographic analysis further showed elevated activation of MMP-2 and MMP-9 following CV133 infection. TIMP-3 and TIMP-4 expression was down-regulated, while TIMP-1 and TIMP-2 remained constant throughout the infection. Mouse hearts stained with picrosirius red showed an increase in total amount of collagen during the acute phase of infection and disrupted fibrils at later timepoints. Conclusion: After CV133 infection, ECM remodeling is triggered, and this response may involve increased expression and activation of MMPs. (C) 2006 Elsevier Inc. All rights reserved.