Characterization of matrix metalloproteinases produced by rat alveolar macrophages
Characterization of matrix metalloproteinases produced by rat alveolar macrophages
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DOI：
10.1165/ajrcmb.20.6.3483
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发表时间：
1999-06-01
影响因子：
6.4
通讯作者：
Varani, J
中科院分区：
文献类型：
--
作者：
Gibbs, DF;Warner, RL;Varani, J
随附文章中提供的证据（Gibbs, D. F., T, P. Shanley, R. L. Warner, H, S, Murphy, J. Varani, and K. J, Johnson. 1999. 基质金属蛋白酶在巨噬细胞依赖性急性肺损伤模型中的作用：肺泡巨噬细胞作为蛋白酶来源的证据。Am. J. Respir. Cell Mel. Biol. 20:1145-1154）表明肺泡巨噬细胞基质金属蛋白酶（MMP）与大鼠急性肺部炎症的两种模型有关。作为了解哪些特定 MMP 可能参与损伤以及它们如何发挥作用的先决条件，有必要了解存在的酶谱。为此，通过支气管肺泡灌洗从正常大鼠肺中获得肺泡巨噬细胞，将其置于含有或不含各种激动剂的培养物中，并通过多种MMP技术进行评估。鉴定过程涉及通过明胶、β-酪蛋白和κ-弹性蛋白酶谱进行表征，并通过蛋白质印迹/免疫沉淀进行身份确认。通过 Northern blot 评估检测到的 MMP 的信息水平。研究发现，大鼠肺泡巨噬细胞产生低水平的 MMP-2（72-kD 明胶酶 A），在用佛波醇肉豆蔻酸酯乙酸酯、细菌脂多糖或含免疫球蛋白 A 的免疫复合物刺激后，该水平仅适度上调。尽管发现对照细胞很少或不产生 MMP-9（92-kD 明胶酶 B）或 MMP-12（金属蛋白酶），但这两种酶在刺激后均显着上调。在相同的刺激巨噬细胞中，几乎没有针对 I 型胶原的活性（根据蛋白质印迹与 MMP-13 [胶原酶-3] 相关），没有暗示基质溶解素或基质溶解素的活性，并且没有可测量的丝氨酸蛋白酶、弹性蛋白酶和组织蛋白酶 G 的分泌。这些数据表明大鼠肺泡巨噬细胞在促炎条件下合成某些 MMP 的能力，这与它们可能参与炎症的进展一致。急性炎症。
Evidence presented in the accompanying article (Gibbs, D. F., T, P. Shanley, R. L. Warner, H, S, Murphy, J. Varani, and K. J, Johnson. 1999. Role of matrix metalloproteinases in models of macrophage-dependent acute lung injury: evidence for alveolar macrophage as source of proteinases. Am. J. Respir. Cell Mel. Biol. 20:1145-1154) implicates alveolar macrophage matrix metalloproteinases (MMPs) in two models of acute lung inflammation in the rat. As a prerequisite to understanding which specific MMPs might be involved in the injury and how they might function, it was necessary to know the spectrum of enzymes present. To this end, alveolar macrophages were obtained from normal rat lungs by bronchoalveolar lavage, placed in culture with and without various agonists, and assessed by a variety of techniques for MMPs, The identification process involved characterization by gelatin, beta-casein, and kappa-elastin zymography, with confirmation of identity by Western blot/immunoprecipitation. Message levels of detected MMPs were assessed by Northern blot. Rat alveolar macrophages were found to produce a low constitutive level of MMP-2 (72-kD gelatinase A) that was only modestly upregulated following stimulation with phorbol myristate acetate, bacterial lipopolysaccharide, or immunoglobulin A-containing immune complexes. Although control cells were found to produce little or no MMP-9 (92-kD gelatinase B) or MMP-12 (metalloelastase), both enzymes were markedly upregulated upon stimulation. In the same stimulated macrophages there was little activity against type I collagen (associated with MMP-13 [collagenase-3] on the basis of Western blotting), no activity suggestive of stromelysin or matrilysin, and no measurable secretion of the serine proteinases, elastase and cathepsin G. These data demonstrate the ability of rat alveolar macrophages to elaborate certain MMPs under proinflammatory conditions, consistent with their possible involvement in the progression of acute inflammation.