Matrix metalloproteinase-9 in laminae of black walnut extract treated horses correlates with neutrophil abundance
Matrix metalloproteinase-9 in laminae of black walnut extract treated horses correlates with neutrophil abundance
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DOI：
10.1016/j.vetimm.2006.05.010
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发表时间：
2006-10-15
影响因子：
1.8
通讯作者：
Black, Samuel J.
中科院分区：
文献类型：
--
作者：
Loftus, John P.;Belknap, James K.;Black, Samuel J.
我们试图确定中性粒细胞浸润和组织基质金属蛋白酶-9（MMP-9）含量之间是否存在相关性，在蹄叶炎的前驱期和急性期收集的马与水黑核桃心材提取物（BWE）。在白细胞减少症发作时和跛行临床体征发作时，从BWE给药马和对照马的相同时间获得蹄层状组织。薄层切片的嗜中性粒细胞进行筛选，通过免疫组织化学与抗CD 13单克隆抗体和提取物的相同的组织进行了筛选SDS-复性和天然MMP-9的活性，变性和非变性明胶酶谱。还通过RT-qPCR筛选样品的MMP-2和MMP-9基因表达。对照板层没有MMP-9和中性粒细胞，而中性粒细胞和SDS-复性MMP-9的活性检测在板层从BWE治疗的马，并在疾病的急性阶段，在该时间层MMP-9基因表达显着（15倍）升高强烈相关。与此相反，BWE治疗没有显着提高NIMP-2基因或蛋白质的表达在椎板。有趣的是，MMP-9是存在于提取物中的薄层从BWE治疗的马在前驱期和急性期的疾病主要是在酶原的形式，这表明MMP的积累并没有在这些阶段的病理。然而，组织中MMP-9酶原的存在升高将使其易于发生灾难性损伤，如果出现切割调节前肽结构域的条件。(c)2006 Elsevier B. V.保留所有权利。
We sought to determine whether a correlation exists between neutrophil infiltration and tissue matrix metalloproteinase-9 (MMP-9) content in digital laminae collected during the prodromal and acute phases of laminitis in horses treated with an aqueous black walnut heartwood extract (BWE). Hoof laminar tissue was obtained at the onset of leukopenia and at the onset of clinical signs of lameness from BWE-treated horses and at equivalent times from control horses. Thin sections of laminae were screened for neutrophils by immunohistochemistry with an anti-CD13 monoclonal antibody and extracts of the same tissues were screened for SDS-renaturable and native MMP-9 activities by denaturing and non-denaturing gelatin zymography. Samples were also screened for MMP-2 and MMP-9 gene expression by RT-qPCR. Control laminae were devoid of both MMP-9 and neutrophils, whereas neutrophils and SDS-renaturable MMP-9 activity were detected in laminae from BWE-treated horses and were strongly correlated at the acute stage of the disease at which time laminar MMP-9 gene expression was significantly (15-fold) elevated. In contrast, BWE-treatment did not significantly elevate NIMP-2 gene or protein expression in the laminae. Interestingly, MMP-9 that was present in extracts of laminae from BWE-treated horses at both the prodromal and acute stages of the disease was mainly in the zymogen form, suggesting that the accumulation of the MMP did not contribute to pathology during these stages. However, elevated presence of the MMP-9 zymogen in the tissue would predispose it to catastrophic damage should conditions arise that cleave the regulatory propeptide domain. (c) 2006 Elsevier B.V. All rights reserved.