Department of Vascular and Endovascular Surgery, Klinikum rechts der Isar der Technischen Universitaet Muenchen, Munich, Germany.
Institute of Pathology, Klinikum rechts der Isar der Technischen Universitaet Muenchen, Munich, Germany.
Exp Mol Med. 2014 May 16;46(5):e95. doi: 10.1038/emm.2014.20.
Cysteine and aspartic proteases possess high elastolytic activity and might contribute to the degradation of the abdominal aortic aneurysm (AAA) wall. The aim of this study was to analyze, in detail, the proteases (cathepsins B, D, K, L and S, and inhibitor cystatin C) found in human AAA and healthy aortic tissue samples. The vessel walls from AAA patients (n=36) and nonaneurysmal aortae (n=10) were retrieved using conventional surgical repair and autopsy methods. Serum samples from the same AAA patients and 10 healthy volunteers were also collected. Quantitative expression analyses were performed at the mRNA level using real-time reverse transcriptase-PCR (RT-PCR). Furthermore, analyses at the protein level included western blot and immunoprecipitation analyses. Cellular sources of cysteine/aspartic proteases and cystatin C were identified by immunohistochemistry (IHC). All cysteine/aspartic proteases and cystatin C were detected in the AAA and control samples. Using quantitative RT-PCR, a significant increase in expression was observed for cathepsins B (P=0.021) and L (P=0.018), compared with the controls. Cathepsin B and cystatin C were also detected in the serum of AAA patients. Using IHC, smooth muscle cells (SMCs) and macrophages were positive for all of the tested cathepsins, as well as cystatin C; in addition, the lymphocytes were mainly positive for cathepsin B, followed by cathepsins D and S. All cysteine/aspartic proteases analyzed in our study were detected in the AAA and healthy aorta. The highest expression was found in macrophages and SMCs. Consequently, cysteine/aspartic proteases might play a substantial role in AAA.
半胱氨酸天冬氨酸蛋白酶具有很强的弹性蛋白酶活性,可能有助于降解腹主动脉瘤(AAA)壁。本研究旨在详细分析在人 AAA 和健康主动脉组织样本中发现的蛋白酶(组织蛋白酶 B、D、K、L 和 S 以及抑制剂胱抑素 C)。使用传统的手术修复和尸检方法从 AAA 患者(n=36)和非动脉瘤主动脉(n=10)中取出血管壁。还收集了来自同一 AAA 患者和 10 名健康志愿者的血清样本。使用实时逆转录聚合酶链反应(RT-PCR)在 mRNA 水平上进行定量表达分析。此外,在蛋白质水平上的分析包括 Western blot 和免疫沉淀分析。通过免疫组织化学(IHC)鉴定半胱氨酸/天冬氨酸蛋白酶和胱抑素 C 的细胞来源。在 AAA 和对照样本中均检测到所有半胱氨酸/天冬氨酸蛋白酶和胱抑素 C。使用定量 RT-PCR,与对照组相比,组织蛋白酶 B(P=0.021)和 L(P=0.018)的表达显著增加。还在 AAA 患者的血清中检测到组织蛋白酶 B 和胱抑素 C。通过 IHC,平滑肌细胞(SMCs)和巨噬细胞对所有测试的组织蛋白酶以及胱抑素 C 均呈阳性;此外,淋巴细胞主要对组织蛋白酶 B 呈阳性,其次是组织蛋白酶 D 和 S。在我们的研究中分析的所有半胱氨酸天冬氨酸蛋白酶均在 AAA 和健康主动脉中检测到。在巨噬细胞和 SMC 中表达最高。因此,半胱氨酸天冬氨酸蛋白酶可能在 AAA 中发挥重要作用。