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定量聚合酶链反应和非常规血清学方法评估氯米帕明在实验性克氏锥虫感染中的治疗效果。

Quantitative PCR and unconventional serological methods to evaluate clomipramine treatment effectiveness in experimental Trypanosoma cruzi infection.

作者信息

Bazán Paola Carolina, Lo Presti María Silvina, Strauss Mariana, Báez Alejandra Lidia, Miler Noemí, Paglini Patricia Adriana, Rivarola Héctor Walter

机构信息

Centro de estudios e investigación de la enfermedad de Chagas y Leishmaniasis. Cátedra de Física Biomédica, (INICSA), CONICET. Facultad de Ciencias Médicas. Universidad Nacional de Córdoba, Santa Rosa 1085, Córdoba, Argentina.

Centro de estudios e investigación de la enfermedad de Chagas y Leishmaniasis. Cátedra de Física Biomédica, (INICSA), CONICET. Facultad de Ciencias Médicas. Universidad Nacional de Córdoba, Santa Rosa 1085, Córdoba, Argentina.

出版信息

Exp Mol Pathol. 2016 Oct;101(2):274-280. doi: 10.1016/j.yexmp.2016.09.006. Epub 2016 Sep 24.

DOI:10.1016/j.yexmp.2016.09.006
PMID:27677396
Abstract

Clomipramine (CLO), a tricyclic antidepressant drug, has been used for the treatment of mice infected with Trypanosoma cruzi. In this work we evaluated the effectiveness of CLO treatment upon T. cruzi-infected mice in the chronic phase of the experimental infection using Quantitative polymerase chain reaction (qPCR) and recombinant ELISA. Sixty Swiss albino mice were inoculated with 50 trypomastigote forms of T. cruzi (Tulahuen strain). CLO treatment consisted of 5mg/kg/day during 60days by intraperitoneal injection, beginning on day 90 post infection (p.i) when the mice presented electrocardiographic (ECG) alterations compatible with the chronic phase of the disease. The evolution of experimental infection and the treatment efficacy were studied through survival, electrocardiography, serology using a mixture and individual (1, 2, 13, 30, 36 and SAPA) recombinant proteins from epimastigotes and trypomastigotes of T. cruzi; and qPCR on days 180 and 270 p.i. CLO treatment in the chronic phase decreased the parasite load, reduced the levels of antibodies against antigen 13 throughout 270days p.i and reversed the ECG abnormalities in the treated animals, from 100% of the mice with alterations at the beginning of the treatment to only 20% of the mice with alterations by day 270 p.i. This study shows that qPCR and the use of recombinant antigens are more sensitive to evaluate the effectiveness of the treatment and proves that clomipramine may be considered as a new chemotherapy for the chronic phase of the disease.

摘要

氯米帕明(CLO)是一种三环类抗抑郁药物,已被用于治疗感染克氏锥虫的小鼠。在本研究中,我们使用定量聚合酶链反应(qPCR)和重组酶联免疫吸附测定(ELISA)评估了CLO治疗对处于实验性感染慢性期的克氏锥虫感染小鼠的有效性。60只瑞士白化小鼠接种了50个克氏锥虫(图拉亨株)的无鞭毛体形式。CLO治疗包括在感染后第90天(p.i)开始,通过腹腔注射,持续60天,剂量为5mg/kg/天,此时小鼠出现与疾病慢性期相符的心电图(ECG)改变。通过生存率、心电图检查、使用来自克氏锥虫无鞭毛体和有鞭毛体的混合及单个(1、2、13、30、36和SAPA)重组蛋白进行血清学检测,以及在感染后第180天和270天进行qPCR,研究了实验性感染的进展和治疗效果。慢性期的CLO治疗降低了寄生虫载量,在感染后270天内降低了针对抗原13的抗体水平,并逆转了治疗动物的心电图异常,从治疗开始时100%的小鼠有改变,到感染后第270天只有20%的小鼠有改变。本研究表明,qPCR和重组抗原的使用对评估治疗效果更敏感,并证明氯米帕明可被视为该疾病慢性期的一种新的化疗药物。

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