Kiczak L, Tomaszek A, Bania J, Paslawska U, Zacharski M, Janiszewski A, Rybinska I, Dziegiel P, von Haehling S, Ardehali H, Jankowska E A, Ponikowski P
Regional Specialist Hospital in Wroclaw, Research and Development Centre, Wroclaw, Poland.
J Physiol Pharmacol. 2014 Jun;65(3):365-75.
Matrix metalloproteinase 9 (MMP-9) is crucial for physiological tissue repair and pathophysiological myocardial remodeling. The regulation of its functioning has been shown to be mediated by formation of complexes with tissue inhibitor of metalloproteinases 1 (TIMP-1) and neutrophil gelatinase associated lipocalin (NGAL). We investigated the mRNA and protein expression of MMP-9, TIMP-1 and NGAL, the formation of complexes, their gelatinolytic activity and cellular localization in left ventricle (LV) from 10 female pigs with induced systolic heart failure (HF), 5 control pigs, and a woman with severe HF. The MMP-9, TIMP-1 and NGAL mRNA in LV did not differ between diseased and healthy pigs. In all pigs MMP-9, TIMP-1 and NGAL proteins were present in LV as high molecular weight (HMW) complexes (115, 130, 170 and 220 kDa), and no monomers were found. A 80 and 115 kDa gelatinolytically active bands were present in all LV homogenates. A 130-kDa active band was seen only in LV from pigs with severe HF. Similar results were found in the explanted heart of a female patient with severe HF. The incubation of the homogenates of porcine LV at 37°C resulted in appearance of 88 kDa active band, which was accompanied by a decreased intensity of HMW bands. The incubation of the homogenates of porcine LV (depleted of active MMP-9) with trypsin generated 80 and 115 kDa active bands. Immunohistochemistry revealed the presence of MMP-9 in the cytoplasm of porcine cardiomyocytes, but not in cardiofibroblasts. Our data suggest that MMP-9 originates from cardiomyocytes, forms the gelatinolytically inactive complexes with TIMP-1 and NGAL, present in normal and failing myocardium, likely serving as a reservoir of active MMP-9. Further studies are needed to elucidate the role of these HMW complexes in the extracellular matrix remodeling during the progression of HF, which presence should be considered when developing efficient strategies inhibiting myocardial matrix metalloproteinases.
基质金属蛋白酶9（MMP - 9）对于生理性组织修复和病理性心肌重塑至关重要。其功能调节已被证明是通过与金属蛋白酶组织抑制剂1（TIMP - 1）和中性粒细胞明胶酶相关脂质运载蛋白（NGAL）形成复合物来介导的。我们研究了10只诱导性收缩性心力衰竭（HF）的雌性猪、5只对照猪以及1名严重HF女性患者左心室（LV）中MMP - 9、TIMP - 1和NGAL的mRNA和蛋白表达、复合物的形成、它们的明胶酶活性以及细胞定位。患病猪和健康猪左心室中的MMP - 9、TIMP - 1和NGAL mRNA没有差异。在所有猪中，MMP - 9、TIMP - 1和NGAL蛋白以高分子量（HMW）复合物（115、130、170和220 kDa）的形式存在于左心室中，未发现单体。所有左心室匀浆中均存在80和115 kDa的具有明胶酶活性的条带。仅在患有严重HF的猪的左心室中可见130 kDa的活性条带。在一名患有严重HF的女性患者的移植心脏中也发现了类似结果。将猪左心室匀浆在37°C孵育导致出现88 kDa的活性条带，同时HMW条带的强度降低。用胰蛋白酶孵育猪左心室匀浆（去除活性MMP - 9）产生了80和115 kDa的活性条带。免疫组织化学显示MMP - 9存在于猪心肌细胞的细胞质中，但不存在于心成纤维细胞中。我们的数据表明，MMP - 9起源于心肌细胞，与TIMP - 1和NGAL形成无明胶酶活性的复合物，存在于正常和衰竭心肌中，可能作为活性MMP - 9的储存库。需要进一步研究以阐明这些HMW复合物在HF进展过程中细胞外基质重塑中的作用，在制定抑制心肌基质金属蛋白酶的有效策略时应考虑到它们的存在。
