Clements J M, Cossins J A, Wells G M, Corkill D J, Helfrich K, Wood L M, Pigott R, Stabler G, Ward G A, Gearing A J, Miller K M
Neures Limited, Abingdon, Oxon, UK.
J Neuroimmunol. 1997 Apr;74(1-2):85-94. doi: 10.1016/s0165-5728(96)00210-x.
Matrix metalloproteinases (MMPs) are a large family of Zn2+ endopeptidases that are expressed in inflammatory conditions and are capable of degrading connective tissue macromolecules. MMP-like enzymes are also involved in the processing of a variety of cell surface molecules including the pro-inflammatory cytokine TNF-alpha. MMPs and TNF-alpha have both been implicated in the pathology associated with neuro-inflammatory diseases (NIDs), particularly multiple sclerosis (MS) and its animal model experimental autoimmune encephalomyelitis (EAE). We have shown that BB-1101, a broad spectrum hydroxamic acid-based combined inhibitor of MMP activity and TNF processing, reduces the clinical signs and weight loss in an acute EAE model in Lewis rats. However, little is known about which MMPs are involved in the neuroinflammatory process. In order to determine the optimum inhibitory profile for an MMP inhibitor in the treatment of NID, we investigated the profile of MMP expression and activity during EAE. The development of disease symptoms was associated with a 3-fold increase in MMP activity in the cerebrospinal fluid (CSF), which could be inhibited by treatment with BB-1101, and an increase in 92 kDa gelatinase activity detected by gelatin substrate zymography. Quantitative PCR analysis of normal and EAE spinal cord revealed the expression of at least seven MMPs. Of these, matrilysin showed the most significant change, being elevated over 500 fold with onset of clinical symptoms and peaking at maximum disease severity. Of the other six MMPs detected, 92 kDa gelatinase showed a modest 5 fold increase which peaked at the onset of clinical signs and then declined during the most severe phase of the disease. Matrilysin was localised by immunohistochemistry to the invading macrophages within the inflammatory lesions of the spinal cord. Matrilysin's potent broad spectrum proteolytic activity and its localisation to inflammatory lesions in the CNS suggest this enzyme could be particularly involved in the pathological processes associated with neuro-inflammatory disease.
基质金属蛋白酶（MMPs）是一个庞大的锌离子内肽酶家族，在炎症条件下表达，能够降解结缔组织大分子。MMP样酶还参与多种细胞表面分子的加工，包括促炎细胞因子肿瘤坏死因子-α（TNF-α）。MMPs和TNF-α都与神经炎症性疾病（NIDs）相关的病理过程有关，特别是多发性硬化症（MS）及其动物模型实验性自身免疫性脑脊髓炎（EAE）。我们已经表明，BB-1101是一种基于异羟肟酸的MMP活性和TNF加工的广谱联合抑制剂，可以减轻Lewis大鼠急性EAE模型中的临床症状和体重减轻。然而，关于哪些MMPs参与神经炎症过程知之甚少。为了确定MMP抑制剂治疗NID的最佳抑制谱，我们研究了EAE期间MMP表达和活性的谱。疾病症状的发展与脑脊液（CSF）中MMP活性增加3倍有关，这可以通过BB-1101治疗来抑制，并且通过明胶底物酶谱法检测到92 kDa明胶酶活性增加。对正常和EAE脊髓的定量PCR分析显示至少七种MMPs的表达。其中，基质溶素显示出最显著的变化，随着临床症状的出现升高超过500倍，并在疾病最严重程度时达到峰值。在检测到的其他六种MMPs中，92 kDa明胶酶显示适度增加5倍，在临床症状开始时达到峰值，然后在疾病最严重阶段下降。通过免疫组织化学将基质溶素定位到脊髓炎症病变内浸润的巨噬细胞。基质溶素强大的广谱蛋白水解活性及其在中枢神经系统炎症病变中的定位表明该酶可能特别参与与神经炎症性疾病相关的病理过程。
