Evaluation of Leishmania infantum 30x biotherapy effects in the prevention and treatment of visceral leishmaniasis: in vivo and in vitro studies

Authors

  • Ana Paula Bacellar Cajueiro UFRJ
  • Gleyce Moreno Barbosa UFRJ
  • Fortune Homsani UFRJ
  • Ana Paula dos Santos Matos UFRJ
  • Igor Almeida Rodrigues UFRJ
  • Leoni Villano Bonamin UNIP
  • Silvana Marques Araújo UEM
  • Katia Fialho do Nascimento UFPR
  • Dorly de Freitas Buchi UFPR
  • Nelson Brêtas de Noronha Gomes UFLA
  • Ester Puna Puna Goma UFRJ
  • Hilton Antônio Mata dos Santos UFRJ
  • Morgana Teixeira Castelo-Branco UFRJ
  • Helena Keiko Toma UFRJ
  • Adriana Passos Oliveira UFRJ
  • Alexandre dos Santos Pyrrho UFRJ
  • Carla Holandino UFRJ

Keywords:

Leishmania infantum, Antimonium crudum, visceral leishmaniasis, biotherapic, nitric oxide.

Abstract

Background: Leishmaniasis is a serious public health problem especially in developing countries [1]. The therapeutic potential of biotherapics against several microorganism has been described in vitro [2,3] and in vivo studies [4,5,6,7,8,9]. Considering the resistance of leishmaniasis to conventional treatment as well as previous studies with biotherapic, we evaluated the effects of Leishmania infantum 30x (BioLi30x) biotherapy. Aim: evaluate the antileishmanial effects of BioLi30x in in vivo and in vitro models. Methodology: The in vivo experiments were performed using BALB/c mice (n=138), divided into 8 groups: G1-healthy, G2-infected with L. infantum, G3-BioLi30x pre-treated, G4-BioLi30x pre/post-treated, G5-BioLi30x post-treated, G6-H2O30x post-treated, G7-Antimonium crudum 30x post-treated and G8-Glucantime® post-treated. After 49 days of treatment, the animals were submitted to euthanasia (ethical approval ECUA/UFRJ/066/14). Liver and spleen histological changes were evaluated, and serum samples were aliquoted and storage at -20°C for cytokine assays. The in vitro assays were performed using RAW 264.7 macrophages treated with BioLi30x and infected with L. infantum. The morphological aspects were evaluated by scanning electron microscopy (SEM), and the nitric oxide (NO) release was quantified in the supernatant of infected macrophages. Results: The histological analysis from 4 independent experiments showed livers with normal appearance (G1); periportal chronic hepatitis (G2,G4,G5,G8); discreet (G3,G7), moderate (G4,G5,G6), and severe (G2,G8) vacuolar hydropic degeneration; congestion and neutrophilic inflammation (G2,G4,G5,G6,G8), and possible amastigotes within macrophages (G2-G8). Spleens presented healthy appearance only in G1. All treated animals presented histological alterations, with different lesions severity, which involved spleen pulp hyperplasia with moderate disruption (G2,G8), as well as megakaryocytes and macrophages proliferation (G2- G8). SEM analyses showed BioLi30x treatments induced significant protozoan morphology alterations when compared to H2O30x. Besides, a 19% increase in the NO release was detected in RAW supernatants, when compared to H2O30x. Conclusions: BioLi30x and Antimonium crudum 30x modified the infection animal process, involving several cellular mechanisms as well as different histological damage. The in vitro experiments will be repeated in order to confirm these preliminary results.

Author Biographies

Ana Paula Bacellar Cajueiro, UFRJ

Multidisciplinary Laboratory of Pharmaceutical Sciences and Laboratory of Research and Development of Integrative and Complementary Medicine, Department of Drugs and Medicines, Pharmacy college, UFRJ. Rio de Janeiro, Brazil – http://www.farmacia.ufrj.br

Gleyce Moreno Barbosa, UFRJ

Multidisciplinary Laboratory of Pharmaceutical Sciences and Laboratory of Research and Development of Integrative and Complementary Medicine, Department of Drugs and Medicines, Pharmacy college, UFRJ. Rio de Janeiro, Brazil – http://www.farmacia.ufrj.br

Fortune Homsani, UFRJ

Multidisciplinary Laboratory of Pharmaceutical Sciences and Laboratory of Research and Development of Integrative and Complementary Medicine, Department of Drugs and Medicines, Pharmacy college, UFRJ. Rio de Janeiro, Brazil – http://www.farmacia.ufrj.br

Ana Paula dos Santos Matos, UFRJ

Multidisciplinary Laboratory of Pharmaceutical Sciences and Laboratory of Research and Development of Integrative and Complementary Medicine, Department of Drugs and Medicines, Pharmacy college, UFRJ. Rio de Janeiro, Brazil – http://www.farmacia.ufrj.br

Igor Almeida Rodrigues, UFRJ

Bioprospecting Laboratory of Natural Antimicrobials, Department of Natural Products and Food, Pharmacy college, UFRJ. Rio de Janeiro, Brazil – http://www.farmacia.ufrj.br

Leoni Villano Bonamin, UNIP

Department of General Pathology and Pathogenesis of susceptibility, UNIP. São Paulo, Brazil - http://www.unip.br

Silvana Marques Araújo, UEM

Department of Basic Health Sciences, UEM. Paraná, Brazil – http://www.uem.br

Katia Fialho do Nascimento, UFPR

Laboratory of Inflammatory and Neoplasic Cells, Department of Cell Biology, UFPR. Paraná, Brazil – http://www.ufpr.br

Dorly de Freitas Buchi, UFPR

Laboratory of Inflammatory and Neoplasic Cells, Department of Cell Biology, UFPR. Paraná, Brazil – http://www.ufpr.br

Nelson Brêtas de Noronha Gomes, UFLA

Department of Veterinary Medicine, UFLA. Minas Gerais, Brazil – http://www.ufla.br

Ester Puna Puna Goma, UFRJ

Laboratory of Immuno-parasitology and Toxicological Analysis, Department of Social Hygiene and Clinical Analysis, Pharmacy college, UFRJ. Rio de Janeiro, Brazil – http://www.farmacia.ufrj.br

Hilton Antônio Mata dos Santos, UFRJ

Laboratory of Immuno-parasitology and Toxicological Analysis, Department of Social Hygiene and Clinical Analysis, Pharmacy college, UFRJ. Rio de Janeiro, Brazil – http://www.farmacia.ufrj.br

Morgana Teixeira Castelo-Branco, UFRJ

Institute of Biomedical Sciences, Department of Histology and Embryology, UFRJ. Rio de Janeiro, Brazil –http://www.icb.ufrj.br

Helena Keiko Toma, UFRJ

Laboratory of Microbiological Control of Drugs, Food and Cosmetics. Department of Clinical and Toxicological Analysis, Pharmacy college, UFRJ. Rio de Janeiro, Brazil – http://www.farmacia.ufrj.br

Adriana Passos Oliveira, UFRJ

Multidisciplinary Laboratory of Pharmaceutical Sciences and Laboratory of Research and Development of Integrative and Complementary Medicine, Department of Drugs and Medicines, Pharmacy college, UFRJ. Rio de Janeiro, Brazil – http://www.farmacia.ufrj.br

Alexandre dos Santos Pyrrho, UFRJ

Laboratory of Immuno-parasitology and Toxicological Analysis, Department of Social Hygiene and Clinical Analysis, Pharmacy college, UFRJ. Rio de Janeiro, Brazil – http://www.farmacia.ufrj.br

Carla Holandino, UFRJ

Multidisciplinary Laboratory of Pharmaceutical Sciences and Laboratory of Research and Development of Integrative and Complementary Medicine, Department of Drugs and Medicines, Pharmacy college, UFRJ. Rio de Janeiro, Brazil – http://www.farmacia.ufrj.br

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Published

2016-11-17

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