Teixeira P C, Iwai L K, Kuramoto A C K, Honorato R, Fiorelli A, Stolf N, Kalil J, Cunha-Neto E
Laboratório de Imunologia, Instituto do Coração, Hospital das Clínicas, Faculdade de Medicina, Universidade de São Paulo, 05403-000 São Paulo, SP, Brazil.
Braz J Med Biol Res. 2006 Dec;39(12):1549-62. doi: 10.1590/s0100-879x2006001200005.
Chronic Chagas' disease cardiomyopathy (CCC) is an often fatal outcome of Trypanosoma cruzi infection, with a poorer prognosis than other cardiomyopathies. CCC is refractory to heart failure treatments, and is the major indication of heart transplantation in Latin America. A diffuse myocarditis, plus intense myocardial hypertrophy, damage and fibrosis, in the presence of very few T. cruzi forms, are the histopathological hallmarks of CCC. To gain a better understanding of the pathophysiology of CCC, we analyzed the protein profile in the affected CCC myocardium. Homogenates from left ventricular myocardial samples of end-stage CCC hearts explanted during heart transplantation were subjected to two-dimensional electrophoresis with Coomassie blue staining; protein identification was performed by MALDI-ToF mass spectrometry and peptide mass fingerprinting. The identification of selected proteins was confirmed by immunoblotting. We demonstrated that 246 proteins matched in gels from two CCC patients. They corresponded to 112 distinct proteins. Along with structural/contractile and metabolism proteins, we also identified proteins involved in apoptosis (caspase 8, caspase 2), immune system (T cell receptor ss chain, granzyme A, HLA class I) and stress processes (heat shock proteins, superoxide dismutases, and other oxidative stress proteins). Proteins involved in cell signaling and transcriptional factors were also identified. The identification of caspases and oxidative stress proteins suggests the occurrence of active apoptosis and significant oxidative stress in CCC myocardium. These results generated an inventory of myocardial proteins in CCC that should contribute to the generation of hypothesis-driven experiments designed on the basis of the classes of proteins identified here.
慢性查加斯病性心肌病（CCC）是克氏锥虫感染常导致的致命后果，其预后比其他心肌病更差。CCC对心力衰竭治疗无效，是拉丁美洲心脏移植的主要指征。弥漫性心肌炎，加上严重的心肌肥大、损伤和纤维化，同时存在极少的克氏锥虫形态，是CCC的组织病理学特征。为了更好地理解CCC的病理生理学，我们分析了受影响的CCC心肌中的蛋白质谱。对心脏移植期间切除的终末期CCC心脏的左心室心肌样本匀浆进行考马斯亮蓝染色二维电泳；通过基质辅助激光解吸电离飞行时间质谱和肽质量指纹图谱进行蛋白质鉴定。通过免疫印迹确认所选蛋白质的鉴定。我们证明，两名CCC患者凝胶中的246种蛋白质相匹配。它们对应于112种不同的蛋白质。除了结构/收缩和代谢蛋白外，我们还鉴定了参与凋亡（半胱天冬酶8、半胱天冬酶2）、免疫系统（T细胞受体β链、颗粒酶A、HLA I类）和应激过程（热休克蛋白、超氧化物歧化酶和其他氧化应激蛋白）的蛋白质。还鉴定了参与细胞信号传导和转录因子的蛋白质。半胱天冬酶和氧化应激蛋白的鉴定表明CCC心肌中存在活跃的凋亡和显著的氧化应激。这些结果生成了CCC心肌蛋白清单，应有助于开展基于此处鉴定的蛋白质类别设计的假设驱动实验。
