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Brazilian Green Propolis: Effects In Vitro and In Vivo on Trypanosoma cruzi.

Salomão K, de Souza EM, Henriques-Pons A, Barbosa HS, de Castro SL - Evid Based Complement Alternat Med (2011)

Bottom Line: These effects were confirmed by flow cytometry analysis.The extract (25-300 mg kg(-1) body weight/day for 10 days) reduced the parasitemia, although not at significant levels; increased the survival of the animals and did not induce any hepatic, muscular lesion or renal toxicity.Et-Bra could be a potential metacyclogenesis blocker, considering its effect on reservosomes, which are an important energy source during parasite differentiation.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Biologia Celular, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Av. Brasil 4365, Manguinhos 21045-900, Rio de Janeiro, Brazil.

ABSTRACT
The composition of a Brazilian green propolis ethanolic extract (Et-Bra) and its effect on Trypanosoma cruzi trypomastigotes and other pathogenic microorganisms have already been reported. Here, we further investigated Et-Bra targets in T. cruzi and its effect on experimental infection of mice. The IC(50)/4 days for inhibition of amastigote proliferation was 8.5 ± 1.8 μg mL(-1), with no damage to the host cells. In epimastigotes Et-Bra induced alterations in reservosomes, Golgi complex and mitochondrion. These effects were confirmed by flow cytometry analysis. In trypomastigotes, Et-Bra led to the loss of plasma membrane integrity. The in vitro studies indicate that Et-Bra interferes in the functionality of the plasma membrane in trypomastigotes and of reservosomes and mitochondrion in epimastigotes. Acutely infected mice were treated orally with Et-Bra and the parasitemia, mortality and GPT, GOT, CK and urea levels were monitored. The extract (25-300 mg kg(-1) body weight/day for 10 days) reduced the parasitemia, although not at significant levels; increased the survival of the animals and did not induce any hepatic, muscular lesion or renal toxicity. Since Et-Bra was not toxic to the animals, it could be assayed in combination with other drugs. Et-Bra could be a potential metacyclogenesis blocker, considering its effect on reservosomes, which are an important energy source during parasite differentiation.

No MeSH data available.


Related in: MedlinePlus

Transmission electron microscopy (a–d) and scanning electron microscopy (e–g) of T. cruzi trypomastigotes treated with Et-Bra for 24 h: (a) control parasite with the normal morphology of mitochondrion (M), kinetoplast (k) and flagellum (f); (b) 30 μg mL−1 and (c, d) 60 μg mL−1 induced the formation of blebs on the body (arrow) and flagellar (arrowheads) membranes. Bars: 0.5 μm. (e) control parasite with the normal morphology; (f, g) 60 μg mL−1 leading formation of blebs on the body (arrow) and flagellar (arrowheads) membranes and some rounded parasites (small arrow). Bars: 4 μm.
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fig4: Transmission electron microscopy (a–d) and scanning electron microscopy (e–g) of T. cruzi trypomastigotes treated with Et-Bra for 24 h: (a) control parasite with the normal morphology of mitochondrion (M), kinetoplast (k) and flagellum (f); (b) 30 μg mL−1 and (c, d) 60 μg mL−1 induced the formation of blebs on the body (arrow) and flagellar (arrowheads) membranes. Bars: 0.5 μm. (e) control parasite with the normal morphology; (f, g) 60 μg mL−1 leading formation of blebs on the body (arrow) and flagellar (arrowheads) membranes and some rounded parasites (small arrow). Bars: 4 μm.

Mentions: Epimastigotes and trypomastigotes were treated for 24 h with Et-Bra and analyzed by transmission (TEM) and scanning electron microscopy (SEM) (Figures 2, 3 and 4). In epimastigotes, the extract caused disorganization of the reservosome morphology (Figures 2(b) and 2(c)), revealed by the reduction of matrix electron density and the increase of both its volume and number of lipid droplets, inducing a heterogeneous arrangement of the organelle and the formation of a crystalloid structure (Figure 2(b)); dilatation of the Golgi complex cisternae (Figure 2(d)); mitochondrial swelling with scarcity of matrix and cristae and the presence of membrane structures inside the organelle (Figures 2(e)–2(h)). By SEM, alterations in epimastigote morphology, shortening and rounding of the parasite's body, (Figures 3(b)–3(d)) were observed. Above 400 μg mL−1, Et-Bra induced an intense vacuolization, preventing the identification of intracellular organelles and leading to the loss of the typical morphology of epimastigotes observed by TEM and SEM (data not shown). Treatment of trypomastigotes caused irregular expansions (blebs) of the body and flagellar membranes (Figures 4(b)–4(d)), confirmed by SEM analysis (Figures 4(f)–4(g)).


Brazilian Green Propolis: Effects In Vitro and In Vivo on Trypanosoma cruzi.

Salomão K, de Souza EM, Henriques-Pons A, Barbosa HS, de Castro SL - Evid Based Complement Alternat Med (2011)

Transmission electron microscopy (a–d) and scanning electron microscopy (e–g) of T. cruzi trypomastigotes treated with Et-Bra for 24 h: (a) control parasite with the normal morphology of mitochondrion (M), kinetoplast (k) and flagellum (f); (b) 30 μg mL−1 and (c, d) 60 μg mL−1 induced the formation of blebs on the body (arrow) and flagellar (arrowheads) membranes. Bars: 0.5 μm. (e) control parasite with the normal morphology; (f, g) 60 μg mL−1 leading formation of blebs on the body (arrow) and flagellar (arrowheads) membranes and some rounded parasites (small arrow). Bars: 4 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3094871&req=5

fig4: Transmission electron microscopy (a–d) and scanning electron microscopy (e–g) of T. cruzi trypomastigotes treated with Et-Bra for 24 h: (a) control parasite with the normal morphology of mitochondrion (M), kinetoplast (k) and flagellum (f); (b) 30 μg mL−1 and (c, d) 60 μg mL−1 induced the formation of blebs on the body (arrow) and flagellar (arrowheads) membranes. Bars: 0.5 μm. (e) control parasite with the normal morphology; (f, g) 60 μg mL−1 leading formation of blebs on the body (arrow) and flagellar (arrowheads) membranes and some rounded parasites (small arrow). Bars: 4 μm.
Mentions: Epimastigotes and trypomastigotes were treated for 24 h with Et-Bra and analyzed by transmission (TEM) and scanning electron microscopy (SEM) (Figures 2, 3 and 4). In epimastigotes, the extract caused disorganization of the reservosome morphology (Figures 2(b) and 2(c)), revealed by the reduction of matrix electron density and the increase of both its volume and number of lipid droplets, inducing a heterogeneous arrangement of the organelle and the formation of a crystalloid structure (Figure 2(b)); dilatation of the Golgi complex cisternae (Figure 2(d)); mitochondrial swelling with scarcity of matrix and cristae and the presence of membrane structures inside the organelle (Figures 2(e)–2(h)). By SEM, alterations in epimastigote morphology, shortening and rounding of the parasite's body, (Figures 3(b)–3(d)) were observed. Above 400 μg mL−1, Et-Bra induced an intense vacuolization, preventing the identification of intracellular organelles and leading to the loss of the typical morphology of epimastigotes observed by TEM and SEM (data not shown). Treatment of trypomastigotes caused irregular expansions (blebs) of the body and flagellar membranes (Figures 4(b)–4(d)), confirmed by SEM analysis (Figures 4(f)–4(g)).

Bottom Line: These effects were confirmed by flow cytometry analysis.The extract (25-300 mg kg(-1) body weight/day for 10 days) reduced the parasitemia, although not at significant levels; increased the survival of the animals and did not induce any hepatic, muscular lesion or renal toxicity.Et-Bra could be a potential metacyclogenesis blocker, considering its effect on reservosomes, which are an important energy source during parasite differentiation.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Biologia Celular, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Av. Brasil 4365, Manguinhos 21045-900, Rio de Janeiro, Brazil.

ABSTRACT
The composition of a Brazilian green propolis ethanolic extract (Et-Bra) and its effect on Trypanosoma cruzi trypomastigotes and other pathogenic microorganisms have already been reported. Here, we further investigated Et-Bra targets in T. cruzi and its effect on experimental infection of mice. The IC(50)/4 days for inhibition of amastigote proliferation was 8.5 ± 1.8 μg mL(-1), with no damage to the host cells. In epimastigotes Et-Bra induced alterations in reservosomes, Golgi complex and mitochondrion. These effects were confirmed by flow cytometry analysis. In trypomastigotes, Et-Bra led to the loss of plasma membrane integrity. The in vitro studies indicate that Et-Bra interferes in the functionality of the plasma membrane in trypomastigotes and of reservosomes and mitochondrion in epimastigotes. Acutely infected mice were treated orally with Et-Bra and the parasitemia, mortality and GPT, GOT, CK and urea levels were monitored. The extract (25-300 mg kg(-1) body weight/day for 10 days) reduced the parasitemia, although not at significant levels; increased the survival of the animals and did not induce any hepatic, muscular lesion or renal toxicity. Since Et-Bra was not toxic to the animals, it could be assayed in combination with other drugs. Et-Bra could be a potential metacyclogenesis blocker, considering its effect on reservosomes, which are an important energy source during parasite differentiation.

No MeSH data available.


Related in: MedlinePlus