Identification of a serum gelatinase associated with the occurrence of cerebral aneurysms as pro-matrix metalloproteinase-2.
Identification of a serum gelatinase associated with the occurrence of cerebral aneurysms as pro-matrix metalloproteinase-2.
复制标题
鉴定与脑动脉瘤发生相关的血清明胶酶为前基质金属蛋白酶-2。
DOI：
10.1161/01.str.29.8.1580
复制
发表时间：
1998
期刊：
影响因子：
8.3
通讯作者：
Chyatte,D
中科院分区：
文献类型：
--
作者：
Todor,DR;Lewis,I;Bruno,G;Chyatte,D
背景和目的--颅内动脉瘤破裂引起的蛛网膜下腔出血是一种严重的中风。细胞外基质重塑与脑动脉瘤有关，并可能在这些病变的形成或破裂中发挥作用。我们之前曾报道，在一组动脉瘤患者中，72 kDa血清明胶酶增加了3倍。本研究的目的是进一步研究和鉴定这种明胶酶。方法：采集手术患者的血清样本。接下来的一系列实验旨在进一步表征和鉴定明胶酶活性增强的患者亚组中发现的主要血清明胶酶。对本地血清样本进行明胶酶谱分析，并与已知的金属蛋白酶激活剂(4-氨基苯基汞醋酸酯[APMA])预处理的血清进行比较。明胶酶谱在基质金属蛋白酶(MMP)抑制剂(EDTA)和丝氨酸蛋白酶抑制剂(苯甲基磺酰氟[PMSF])存在下重复进行。用抗基质金属蛋白酶-2和基质金属蛋白酶-9的单抗进行Western blotting鉴定。结果：60份血清标本均有一条一致的明胶溶解条带(72 KDa)。用APMA(n=60)对血清进行预处理，使该条带的相对分子质量降至66 kDa。该条带被EDTA(n=10)抑制，但不被PMSF(n=10)抑制，因此循环中的72-kDa明胶酶是一种无活性的前-基质金属蛋白酶。Western blotting(n=20)鉴定72 kDa条带为基质金属蛋白酶-2。结论--这些发现证实了在体外观察到的部分脑动脉瘤患者的明胶溶解活性增加是由于基质金属蛋白酶-2原。
Background and Purpose—Subarachnoid hemorrhage from intracranial aneurysm rupture produces a severe form of stroke. Extracellular matrix remodeling is associated with cerebral aneurysms and may play a role in the formation or rupture of these lesions. We previously reported a 3-fold increase in a 72-kDa serum gelatinase in a subgroup of aneurysm patients. The purpose of the present study was to further characterize and identify this gelatinase.Methods—Serum samples were collected from surgical patients with intracranial aneurysms. The following series of experiments was designed to further characterize and identify the predominant serum gelatinase found in the subgroup of patients with increased gelatinase activity. Gelatin zymography was performed on native serum samples and compared with serum that had been pretreated with a known metalloproteinase activator (4-aminophenylmercuric acetate [APMA]). Gelatin zymography was repeated in the presence of a matrix metalloproteinase (MMP) inhibitor (EDTA) and a serine proteinase inhibitor (phenylmethylsulfonyl fluoride [PMSF]). Final identification was made by Western blotting with the use of monoclonal antibodies to MMP-2 and MMP-9.Results—A consistent gelatinolytic band (72 kDa) was identified in all serum samples (n=60). Pretreatment of the serum by APMA (n=60) lowered the molecular weight of the band to 66 kDa. The band was inhibited by EDTA (n=10) but not PMSF (n=10), thus characterizing the circulating 72-kDa gelatinase as an inactive pro-MMP. Western blotting (n=20) identified the 72-kDa band as MMP-2.Conclusions—These findings confirm that the increased gelatinolytic activity observed in vitro in a subset of cerebral aneurysm patients is due to pro-MMP-2.