Mendonça Débora V C, Lage Letícia M R, Lage Daniela P, Chávez-Fumagalli Miguel A, Ludolf Fernanda, Roatt Bruno M, Menezes-Souza Daniel, Faraco André A G, Castilho Rachel O, Tavares Carlos A P, Barichello José Mário, Duarte Mariana C, Coelho Eduardo A F
Programa de Pós-Graduação em Ciências da Saúde: Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
Departamento de Patologia Clínica, COLTEC, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
Exp Parasitol. 2016 Oct;169:34-42. doi: 10.1016/j.exppara.2016.07.005. Epub 2016 Jul 15.
In the present study, a Poloxamer 407-based amphotericin B (AmpB)-containing polymeric micelles system (AmpB/M) was employed in the treatment of Leishmania amazonensis-infected BALB/c mice. Initially, the in vitro antileishmanial activity (IC50 value) of AmpB/M and B-AmpB/M (empty micelles) against stationary promastigotes and amastigotes-like forms of the parasites was determined, and results were of 1.83 ± 0.4 and 22.1 ± 0.7 μM, respectively, for the promastigotes, and of 2.27 ± 0.5 and 33.98 ± 2.6 μM, respectively, for the amastigotes-like. The cytotoxic concentration (CC50) values of these products were also evaluated, and we found the results of 119.5 ± 9.6 and 134.7 ± 10.3 μM, respectively. With these values, the selectivity index (SI) was calculated and results were of 65.3 and 5.4, respectively, for the promastigotes, and of 59.3 and 3.96, respectively, for the amastigotes-like of the parasites. Free AmpB showed IC50 values of 1.2 ± 0.3 and 2.5 ± 0.5 μM for the promastigotes and amastigotes-like, respectively, whereas the CC50 value was of 9.5 ± 0.4 μM. The SI values of this drug were of 7.9 and 3.8, respectively, for the promastigote and amastigote-like stages of the parasites. After, animals were infected and received saline or were treated subcutaneously with free AmpB, AmpB/M or B-AmpB/M. In the results, free AmpB-treated and infected mice showed reductions in their body weight, which were associated with hepatic and renal damage; however, no organic alteration was observed in the AmpB/M-treated animals. In addition, these animals showed significant reductions in their lesion average size and in the parasite burden in all evaluated infected tissue and organs, when compared to the other groups; as well as significantly higher levels of antileishmanial IFN-γ, IL-12, GM-CSF and nitrite, which were associated with low production of IL-4, IL-10 and IgG1 isotype antibodies. In conclusion, this AmpB/M system could be considered as an alternative for future studies in the treatment of tegumentary leishmaniasis.
在本研究中，采用了基于泊洛沙姆407的含两性霉素B（AmpB）的聚合物胶束系统（AmpB/M）来治疗感染亚马逊利什曼原虫的BALB/c小鼠。首先，测定了AmpB/M和B - AmpB/M（空胶束）对静止前鞭毛体和类无鞭毛体形式寄生虫的体外抗利什曼原虫活性（IC50值），结果显示，对于前鞭毛体，IC50值分别为1.83±0.4和22.1±0.7 μM，对于类无鞭毛体，IC50值分别为2.27±0.5和33.98±2.6 μM。还评估了这些产品的细胞毒性浓度（CC50）值，我们发现结果分别为119.5±9.6和134.7±10.3 μM。根据这些值，计算出选择性指数（SI），对于前鞭毛体，结果分别为65.3和5.4，对于寄生虫的类无鞭毛体，结果分别为59.3和3.96。游离AmpB对前鞭毛体和类无鞭毛体的IC50值分别为1.2±0.3和2.5±0.5 μM，而CC50值为9.5±0.4 μM。该药物对寄生虫前鞭毛体和类无鞭毛体阶段的SI值分别为7.9和3.8。之后，动物被感染并接受生理盐水，或皮下注射游离AmpB、AmpB/M或B - AmpB/M。结果显示，接受游离AmpB治疗的感染小鼠体重减轻，这与肝损伤和肾损伤有关；然而，在接受AmpB/M治疗的动物中未观察到器官改变。此外，与其他组相比，这些动物在所有评估的感染组织和器官中的病变平均大小和寄生虫负荷均显著降低；同时，抗利什曼原虫的IFN-γ、IL-12、GM-CSF和亚硝酸盐水平显著升高，这与IL-4、IL-10和IgG1同种型抗体的低产生有关。总之，这种AmpB/M系统可被视为未来治疗皮肤利什曼病研究的一种替代方案。
