Ethanolic extract of Passiflora edulis Sims leaves inhibits protein glycation and restores the oxidative burst in diabetic rat macrophages after Candida albicans exposure

This study was conducted to evaluate the effects of the ethanolic extract of Passiflora edulis leaves on blood glucose, protein glycation, NADPH oxidase activity and macrophage phagocytic capacity after Candida albicans exposure in diabetic rats. The Passiflora edulis Sims leaves were dried to 40°C, powdered, extracted by maceration in 70% ethanol, evaporated under reduced pressure and lyophilised. The biochemical tests performed were total phenolic content (TP) as determined by the Folin-Ciocalteu assay, trapping potential DPPH assay and total iron-reducing potential. Diabetes was induced by alloxan injection. Protein glycation was determined by AGE and fructosamine serum concentrations. Extract-treated diabetic animals demonstrated lower fructosamine concentrations compared with the diabetic group. Our results suggest that ethanolic Passiflora edulis Sims leaf extraction may have beneficial effects on diabetes and may improve glycaemic control in diabetic rats.

Uniterms:
Passiflora edulis Sims/pharmacognosy; Passiflora edulis Sims/ethanolic extract/effects; Blood glucose; Protein glycation; Reactive oxygen species; Diabetes/treatment/experimental study; Medicinal plants.

Authorship

Carolina Fernandes Ribas Martins

Faculty of Pharmaceutical Sciences, Departament of Clinical and Toxicology Analysis, Federal University of Alfenas, MG, BrazilFederal University of AlfenasBrazilMG, BrazilFaculty of Pharmaceutical Sciences, Departament of Clinical and Toxicology Analysis, Federal University of Alfenas, MG, Brazil

Bruno César Corrêa Salles

Maisa Ribeiro Pereira Lima Brigagão

Institute of Biomedical Sciences, Department of Biochemistry, Federal University of Alfenas, MG, BrazilFederal University of AlfenasBrazilMG, Brazil Institute of Biomedical Sciences, Department of Biochemistry, Federal University of Alfenas, MG, Brazil

Maria Rita Rodrigues

Eric Batista Ferreira

Insitute of Exact Sciences, Federal University of Alfenas, Federal University of Alfenas, MG, BrazilFederal University of AlfenasBrazilMG, Brazil Insitute of Exact Sciences, Federal University of Alfenas, Federal University of Alfenas, MG, Brazil

Stella Maris da Silveira Duarte

Fernanda Borges de Araujo Paula ^**Correspondence: F. B. de Araujo Paula. Departamento de Análises Clínicas. Faculdade de Ciências Farmacêuticas. Universidade Federal de Alfenas. Rua Gabriel Monteiro da Silva, 700. Centro, 37130-000 - Alfenas - MG, Brasil. E-mail addresses: fbapaula@yahoo.com.br; fernanda.paula@unifal-mg.edu.br

*Correspondence: F. B. de Araujo Paula. Departamento de Análises Clínicas. Faculdade de Ciências Farmacêuticas. Universidade Federal de Alfenas. Rua Gabriel Monteiro da Silva, 700. Centro, 37130-000 - Alfenas - MG, Brasil. E-mail addresses: fbapaula@yahoo.com.br; fernanda.paula@unifal-mg.edu.br

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Figures | Tables

Figures (4)
Tables (1)

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FIGURE 1
Trapping Potential DPPH Assay of the Passiflora edulis leaf ethanolic extract compared with the standard BHT.

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FIGURE 2
Total iron-reducing power assay of the Passiflora edulis leaf ethanolic extract. The results represent the average  standard deviation performed in triplicate.

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FIGURE 3
Effect of P. edulis leaf ethanolic extract on the ability of diabetic and non-diabetic rat peritoneal macrophages to phagocyte Candida albicans. Macrophages were incubated with opsonised yeast C. albicans for 0, 30, 60, 90 or 120 minutes, and the percentage of macrophages that phagocytised one or more yeast molecules (% phagocytosis) was determined. The results represent the average  standard deviation of 4-5 determinations per treatment. C=control group; D=diabetic animals; E=P. edulis ethanolic extract-treated non-diabetic animals; DE=P. edulis ethanolic extract-treated diabetic animals.

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FIGURE 4
Effect of the Passiflora edulis leaf ethanolic extract on Reactive Oxygen Species (ROS) production in diabetic and non-diabetic rats. The values are represented as the means ± SD of the four-five experiments of the integrated light emission area that was obtained for luminol oxidation (1 mmol.L^-1) promoted by Candida albicans (1x10⁷ cells per test)-stimulated rat peritoneal macrophages (1x10⁶ cells per test) from control animals (C), diabetic animals (D), Passiflora edulis ethanolic extract-treated non-diabetic animals (E) and Passiflora edulis ethanolic extract-treated diabetic animals (DE). Different letters indicate statistical significance (p<0.05).

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TABLE I
Effect of P. edulis leaf ethanolic extract on glucose levels, fructosamine concentrations and advanced glycation end-product (AGE) quantification in non-diabetic and diabetic animals

FIGURE 1 Trapping Potential DPPH Assay of the Passiflora edulis leaf ethanolic extract compared with the standard BHT.

FIGURE 2 Total iron-reducing power assay of the Passiflora edulis leaf ethanolic extract. The results represent the average  standard deviation performed in triplicate.

FIGURE 3 Effect of P. edulis leaf ethanolic extract on the ability of diabetic and non-diabetic rat peritoneal macrophages to phagocyte Candida albicans. Macrophages were incubated with opsonised yeast C. albicans for 0, 30, 60, 90 or 120 minutes, and the percentage of macrophages that phagocytised one or more yeast molecules (% phagocytosis) was determined. The results represent the average  standard deviation of 4-5 determinations per treatment. C=control group; D=diabetic animals; E=P. edulis ethanolic extract-treated non-diabetic animals; DE=P. edulis ethanolic extract-treated diabetic animals.

FIGURE 4 Effect of the Passiflora edulis leaf ethanolic extract on Reactive Oxygen Species (ROS) production in diabetic and non-diabetic rats. The values are represented as the means ± SD of the four-five experiments of the integrated light emission area that was obtained for luminol oxidation (1 mmol.L^-1) promoted by Candida albicans (1x10⁷ cells per test)-stimulated rat peritoneal macrophages (1x10⁶ cells per test) from control animals (C), diabetic animals (D), Passiflora edulis ethanolic extract-treated non-diabetic animals (E) and Passiflora edulis ethanolic extract-treated diabetic animals (DE). Different letters indicate statistical significance (p<0.05).

TABLE I Effect of P. edulis leaf ethanolic extract on glucose levels, fructosamine concentrations and advanced glycation end-product (AGE) quantification in non-diabetic and diabetic animals

Experimental group	Glycaemia (mg/dl)	Fructosamines (mmol/L)	AGE (AU/g prot)	Body weight (g)
1^st week	8^st week
C	113,78 ± 13,52^a	1,63 ± 0,18^a	0,11 ± 0,02^a	354,50 ± 20,87^a	417,00 ± 17,03^a
D	285,78 ± 69,35^b	2,38 ± 0,41^b	0,17 ± 0,06^b	351,00 ± 19,34^a	264,67 ± 38,05^b
E	90,95 ± 10,57^a	1,61 ± 0,07^a	0,10 ± 0,02^a	344,17 ± 17,05^a	401,83 ± 22,69^a
DE	269,98 ± 115,58^a	1,85 ± 0,49^a	0,13 ± 0,04^a	348,50 ± 15,75^a	280,66 ± 36,35^b

The results represent the average  standard deviation of 6-8 determinations per treatment. Different letters in the same column indicate statistical significance, p<0.05. C=control group; D=diabetic animals; E=P. edulis ethanolic extract-treated non-diabetic animals; DE=P. edulis ethanolic extract-treated diabetic animals.

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Ethanolic extract of Passiflora edulis Sims leaves inhibits protein glycation and restores the oxidative burst in diabetic rat macrophages after Candida albicans exposure

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[1] *Correspondence: F. B. de Araujo Paula. Departamento de Análises Clínicas. Faculdade de Ciências Farmacêuticas. Universidade Federal de Alfenas. Rua Gabriel Monteiro da Silva, 700. Centro, 37130-000 - Alfenas - MG, Brasil. E-mail addresses: fbapaula@yahoo.com.br; fernanda.paula@unifal-mg.edu.br