MMP-9 regulates both positively and negatively collagen gel contraction - A nonproteolytic function of MMP-9
MMP-9 regulates both positively and negatively collagen gel contraction - A nonproteolytic function of MMP-9
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DOI：
10.1016/j.cardiores.2004.11.025
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发表时间：
2005-05-01
影响因子：
10.8
通讯作者：
Clowes, AW
中科院分区：
文献类型：
--
作者：
Defawe, OD;Kenagy, RD;Clowes, AW
目的：收缩性重塑是血管成形术后再狭窄中管腔损失的原因。基质金属蛋白酶-9 (MMP-9) 已被证明可以防止体内收缩性重塑。为了研究这一观察的潜在机制，我们研究了 MMP-9 在平滑肌细胞 (SMC) 介导的胶原凝胶收缩中的作用，这是一种收缩重塑的体外模型。方法：在四环素 (Tet) 关闭启动子的控制下，用表达大鼠 MMP-9 的构建体稳定转染 Fischer 大鼠 SMC。在存在或不存在四环素（1μg/ml）的情况下将SMC接种于1型胶原凝胶（2.4mg/ml）中，并且凝胶收缩定义为胶原凝胶收缩的百分比。通过使用靶向 MMP-9 mRNA 的 siRNA 或封闭抗体来消除 MMP-9。 结果：与对照条件 (Tet+) 相比，当 MMP-9 过表达 (Tet-) 时，凝胶收缩始终显着减少。然而，对照 (Tet+) SMCS（使用 siRNA 或抗体）的 MMP-9 耗尽也会抑制凝胶收缩。为了解决明显的差异并确定 MMP-9 是否对凝胶收缩产生剂量依赖性双相效应，制备了条件培养基和纯化的大鼠 MMP-9。添加 0.8 mg/ml 的 MMP-9 显着增加凝胶收缩，而高浓度的 MMP-9 (>= 100 mg/ml) 抑制收缩。添加 BB94 和 TIMP-1 不会改变 MMP-9 的抑制或刺激作用。结论：我们的数据表明，MMP-9 独立于其蛋白水解功能，对 SMC 介导的胶原凝胶收缩具有双相作用。了解 MMP-9 的不同作用应该有助于开发更好的再狭窄血管疾病治疗策略。 (c) 2004 年欧洲心脏病学会。由 Elsevier B.V. 出版。保留所有权利。
Objective: Constrictive remodeling accounts for lumen loss in postangioplasty restenosis. Matrix metalloproteinase-9 (MMP-9) has been shown to prevent constrictive remodeling in vivo. To investigate potential mechanisms for this observation, we investigated the role of MMP-9 in smooth muscle cell (SMC)-mediated collagen gel contraction, an in vitro model of constrictive remodeling.Methods: Fischer rat SMCs were stably transfected with a construct-expressing rat-MMP-9 under the control of a tetracycline (Tet)-off promoter. SMCs were seeded in type 1 collagen gels (2.4 mg/ml) in the presence or not of tetracycline (1 mu g/ml), and gel contraction was defined as the percentage of retraction of the collagen gel. The depletion of MMP-9 was obtained by using siRNA targeting MMP-9 mRNA or a blocking antibody.Results: Gel contraction was significantly reduced at all times when MMP-9 was overexpressed (Tet-) as compared with the control condition (Tet+). However, MMP-9 depletion of control (Tet+) SMCS (using siRNA or antibody) also inhibited gel contraction. To resolve the apparent discrepancy and determine if MMP-9 exerts a dose-dependent biphasic effect on gel contraction, conditioned medium and purified rat-MMP-9 were prepared. Gel contraction was significantly increased by addition of 0.8 mg/ml of MMP-9, while high concentrations of MMP-9 (>= 100 mg/ml) inhibited contraction. The addition of BB94 and TIMP-1 did not alter the inhibitory or stimulatory effect of MMP-9.Conclusions: Our data Suggest that MMP-9, independent of its proteolytic function, has a biphasic effect on SMC-mediated collagen gel contraction. Understanding the different roles of MMP-9 Should allow the development of better therapeutic strategies for restenotic vascular disease. (c) 2004 European Society of Cardiology. Published by Elsevier B.V. All rights reserved.