Rodríguez-Angulo Héctor O, Toro-Mendoza Jhoan, Marques Juan A, Concepción Juan L, Bonfante-Cabarcas Rafael, Higuerey Yoliver, Thomas Luz E, Balzano-Nogueira Leandro, López José R, Mijares Alfredo
Laboratorio de Fisiología de Parásitos, Centro de Biofísica y Bioquímica, Instituto Venezolano de Investigaciones Científicas, Caracas, Venezuela.
Centro de Estudios Interdisciplinarios de la Física. Instituto Venezolano de Investigaciones Científicas, Caracas, Venezuela.
PLoS Negl Trop Dis. 2015 Feb 3;9(2):e0003512. doi: 10.1371/journal.pntd.0003512. eCollection 2015 Feb.
Chagas cardiomyopathy, caused by the protozoan Trypanosoma cruzi, is characterized by alterations in intracellular ion, heart failure and arrhythmias. Arrhythmias have been related to sudden death, even in asymptomatic patients, and their molecular mechanisms have not been fully elucidated.
The aim of this study is to demonstrate the effect of proteins secreted by T. cruzi on healthy, isolated beating rat heart model under a non-damage-inducing protocol.
We established a non-damage-inducing recirculation-reoxygenation model where ultrafiltrate fractions of conditioned medium control or conditioned infected medium were perfused at a standard flow rate and under partial oxygenation. Western blotting with chagasic patient serum was performed to determine the antigenicity of the conditioned infected medium fractions. We observed bradycardia, ventricular fibrillation and complete atrioventricular block in hearts during perfusion with >50 kDa conditioned infected culture medium. The preincubation of conditioned infected medium with chagasic serum abolished the bradycardia and arrhythmias. The proteins present in the conditioned infected culture medium of >50 kDa fractions were recognized by the chagasic patient sera associated with arrhythmias.
These results suggest that proteins secreted by T. cruzi are involved in Chagas disease arrhythmias and may be a potential biomarker in chagasic patients.
由原生动物克氏锥虫引起的恰加斯心肌病,其特征为细胞内离子改变、心力衰竭和心律失常。心律失常与猝死有关,即使在无症状患者中也是如此,其分子机制尚未完全阐明。
本研究的目的是在不诱导损伤的方案下,证明克氏锥虫分泌的蛋白质对健康的离体搏动大鼠心脏模型的影响。
我们建立了一个不诱导损伤的再循环 - 复氧模型,以标准流速并在部分氧合条件下灌注条件培养基对照或条件感染培养基的超滤部分。用恰加斯病患者血清进行蛋白质印迹法,以确定条件感染培养基部分的抗原性。在用分子量大于50 kDa的条件感染培养基灌注心脏期间,我们观察到心动过缓、心室颤动和完全性房室传导阻滞。将条件感染培养基与恰加斯病血清预孵育可消除心动过缓和心律失常。分子量大于50 kDa部分的条件感染培养基中存在的蛋白质可被与心律失常相关的恰加斯病患者血清识别。
这些结果表明,克氏锥虫分泌的蛋白质与恰加斯病心律失常有关,可能是恰加斯病患者的潜在生物标志物。