Fig 1.
Liquid chromatography–mass spectrometry analysis of Morinda citrifolia fruit juice, Noni.
(1–5) Chromatograms of compounds (m/z) identified in Noni: deacetylasperulosidic acid (389), asperulosidic acid (431), rutin (610), nonioside B (629) and nonioside C (467).
Fig 2.
Activity of Morinda citrifolia fruit juice (Noni) treatment in C57BL/6 mice infected with Leishmania (L.) amazonensis.
(A) Kinetic of lesion of infected footpads treated with Noni (500mg.kg-1.day-1) or Glucantime (20mg.kg-1.twice a week-1). (B-C) Parasite loads in footpad and draining lymph node after 30 and 60 days of Noni treatment. Data represent mean ± SD of two independent experiments realized at least in triplicate. *p<0.05, **p<0.01, ***p<0.001 when compared with La group by two-way ANOVA and Bonferroni’s post-test. La+Noni: group infected and treated with Noni; La+Glucantime: group infected and treated with Glucantime; La: group infected and mock-treated; Normal: mock-infected and mock-treated group.
Fig 3.
Histopathological analysis of skin and lymph nodes of C57BL/6 mice footpads infected with Leishmania (L.) amazonensis and treated with Morinda citrifolia fruit juice, Noni.
Noni group treated at 500mg.kg-1.day-1 and Glucantime group treated at 20mg.kg-1.twice a week-1; both treatments were for 60 days. At 30 days: amastigotes within macrophages (arrows in inserts) and inflammatory infiltration in dermis of all groups. At 60 days: inflammatory infiltration decreased in the Noni group, and absence of amastigotes in the Noni and Glucantime groups. La group with intense inflammatory infiltration of macrophages with parasites at the lesion site. In lymph nodes, there was a decrease of the lymphoid nodules (asterisks) hyperplasia in Noni and Glucantime groups. Images representative of two experiments realized in triplicate. Hematoxylin-eosin.
Fig 4.
Cytokine gene expression in C57BL/6 mouse footpads infected with Leishmania (L.) amazonensis and treated with Morinda citrifolia fruit juice.
Relative quantification of IFN-γ (A), iNOS (B), IL-12 (C), TNF-α (D), IL-10 (E), TGF-β (F) and IL-4 (G) with RPLP0 as endogenous control. Noni group treated at 500mg.kg-1.day-1 and Glucantime group treated at 20mg.kg-1.twice a week-1, both during 60 days. Data represent mean ± SD of two experiments realized in triplicate. *p<0.05, **p<0.01, ***p<0.001 when compared with control group or between group brackets by two-way ANOVA and Bonferroni’s post-test. RQ: relative quantification; RPLP0: ribosomal protein large P0; La+Noni: group infected and treated with Noni; La+Glucantime: group infected and treated with Glucantime; La: group infected and mock-treated; Noni: group mock-infected and treated with Noni; Normal: mock-infected and mock-treated group.
Fig 5.
Quantification of serum cytokines from C57BL/6 mice infected with Leishmania (L.) amazonensis and treated with Morinda citrifolia fruit juice.
Cytokines levels of IFN-γ (A), IL-12 (B), TNF-α (C), IL-4 (D), IL-10 (E) and TGF-β (F). Noni group treated at 500mg.kg-1.day-1 and Glucantime group treated at 20mg.kg-1.twice a week-1, both for 60 days. Data represent mean ± SD of two experiments realized in duplicate. *p<0.05, ***p<0.001 when compared with control group or between group brackets by two-way ANOVA and Bonferroni’s post-test. La+Noni: group infected and treated with Noni; La+Glucantime: group infected and treated with Glucantime; La: group infected and mock-treated; Noni: group mock-infected and treated with Noni; Normal: mock-infected and mock-treated group.
Fig 6.
Extracellular matrix protein analysis of C57BL/6 mice footpads infected with Leishmania amazonensis and treated for 60 days with Morinda citrifolia fruit juice, Noni.
Noni group treated at 500mg.kg-1.day-1 and Glucantime group treated at 20mg.kg-1.twice a week-1. Histopathology (A) and extracellular matrix protein gene expressions (B-F) of skin. Images are representative of two independent experiments realized in triplicate. HE: hematoxylin-eosin. Data represent mean ± SD of two independent experiments realized in triplicate. *p<0.05, **p<0.01, ***p<0.001 when compared with control group or between group brackets by one-way ANOVA and Bonferroni’s post-test. RQ: relative quantification; RPLP0: ribosomal protein large P0; La+Noni: group infected and treated with Noni; La+Glucantime: group infected and treated with Glucantime; La: group infected and mock-treated; Noni: group mock-infected and treated with Noni; Normal: mock-infected and mock-treated group.
Fig 7.
Quantification of alanine aminotransferase (ALT) and histopathology of liver from C57BL/6 mice infected with Leishmania amazonensis and treated for 60 days with Morinda citrifolia fruit juice, Noni.
Noni group treated at 500mg.kg-1.day-1 and Glucantime group treated at 20mg.kg-1.twice a week-1. Diffuse inflammatory infiltration (arrows) and periportal infiltration (arrow-heads) in the liver of infected mice. The inflammatory intensity of infiltration decreases with Glucantime and Noni treatments. Hematoxylin-eosin. Data represent mean ± SD of two independent experiments realized in duplicate. *p<0.05, **p<0.01 when compared with control group or between group brackets by one-way ANOVA and Bonferroni’s post-test. La+Noni: group infected and treated with Noni; La+Glucantime: group infected and treated with Glucantime; La: group infected and mock-treated; Noni: group mock-infected and treated with Noni; Normal: mock-infected and mock-treated group.