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Evaluation of Antiradical and Anti-Inflammatory Activities of Ethyl Acetate and Butanolic Subfractions of Agelanthus dodoneifolius (DC.) Polhill & Wiens (Loranthaceae) Using Equine Myeloperoxidase and Both PMA-Activated Neutrophils and HL-60 Cells.

Boly R, Franck T, Kohnen S, Lompo M, Guissou IP, Dubois J, Serteyn D, Mouithys-Mickalad A - Evid Based Complement Alternat Med (2015)

Bottom Line: The results showed the potent activity of the butanolic subfraction, at least in respect of the chemiluminescence test (IC50 = 0.3 ± 0.1 µg/mL) and the ELISA and SIEFED assays (IC50 = 2.8 ± 1.2 µg/mL and 1.3 ± 1.0 µg/mL), respectively.However, the ethyl acetate subfraction was found to be the most potent in the DCF assay as at the highest concentration, DCF fluorescence intensity decreases of about 50%.Nevertheless, taken together, our results provide evidence that Agelanthus dodoneifolius subfractions may represent potential sources of natural antioxidants and of antimyeloperoxidase compounds.

View Article: PubMed Central - PubMed

Affiliation: Institute for Research in Health Sciences (IRSS/CNRST), 03 BP 7192 Ouagadougou 03, Burkina Faso.

ABSTRACT
The ethyl acetate and n-butanolic subfractions of Agelanthus dodoneifolius were investigated for their antioxidant and antimyeloperoxidase (MPO) activities. The reactive oxygen species (ROS) generation was assessed by lucigenin-enhanced chemiluminescence (CL) and dichlorofluorescein- (DCF-) induced fluorescence techniques from phorbol myristate acetate- (PMA-) stimulated equine neutrophils and human myeloid cell line HL-60, respectively. In parallel, the effects of the tested subfractions were evaluated on the total MPO release by stimulated neutrophils and on the specific MPO activity by means of immunological assays. The results showed the potent activity of the butanolic subfraction, at least in respect of the chemiluminescence test (IC50 = 0.3 ± 0.1 µg/mL) and the ELISA and SIEFED assays (IC50 = 2.8 ± 1.2 µg/mL and 1.3 ± 1.0 µg/mL), respectively. However, the ethyl acetate subfraction was found to be the most potent in the DCF assay as at the highest concentration, DCF fluorescence intensity decreases of about 50%. Moreover, we demonstrated that the ethyl acetate subfraction was rich in catechin (16.51%) while it was not easy to identify the main compounds in the butanolic subfraction using the UPLC-MS/MS technique. Nevertheless, taken together, our results provide evidence that Agelanthus dodoneifolius subfractions may represent potential sources of natural antioxidants and of antimyeloperoxidase compounds.

No MeSH data available.


Related in: MedlinePlus

UPLC-UV chromatogram profile of the ethyl acetate subfraction acquired at 280 nm with the photodiode array detector. The subfraction was solubilized in methanol and injected onto an Acquity UPLC BEH Shield RP18 column (2.1 × 100 mm, 1.7 μm; Waters). Peaks 1: gallic acid; 2: catechin; 3: unknown; 4: unknown.
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fig3: UPLC-UV chromatogram profile of the ethyl acetate subfraction acquired at 280 nm with the photodiode array detector. The subfraction was solubilized in methanol and injected onto an Acquity UPLC BEH Shield RP18 column (2.1 × 100 mm, 1.7 μm; Waters). Peaks 1: gallic acid; 2: catechin; 3: unknown; 4: unknown.

Mentions: The lowest IC50 value was found for the butanolic subfraction (IC50 = 0.3 ± 0.1 μg/mL) but was statistically similar to that obtained with the ethyl acetate subfraction (IC50 = 0.4 ± 0.2 μg/mL) and gallic acid (IC50 = 0.4 ± 0.1 μg/mL). Lucigenin is a well-known luminescent probe that is widely used for its high specificity towards superoxide radical [26]. Our results strongly suggest that Agelanthus dodoneifolius subfractions are potential scavengers of superoxide radical, arising from the NADPH oxidase activation, and so that constitutes the first step in the generation of most of the other ROS [27]. Polyphenols such as flavonoids are known to inhibit enzymes responsible for superoxide anion production such as xanthine oxidase, protein kinase C, and NADPH oxidase [28]. It has been suggested that the health benefits afforded by the consumption of fruits, legumes, vegetables, and whole grains are mainly due to the presence of natural antioxidants such as ascorbic acid, carotenoids, tocopherols, and phenolic compounds which represent the most important class [25, 28, 29]. The significant effect of the ethyl acetate and n-butanol subfractions is related to their content of polyphenols compounds as demonstrated by the UPLC-MS/MS analysis (Figure 3).


Evaluation of Antiradical and Anti-Inflammatory Activities of Ethyl Acetate and Butanolic Subfractions of Agelanthus dodoneifolius (DC.) Polhill & Wiens (Loranthaceae) Using Equine Myeloperoxidase and Both PMA-Activated Neutrophils and HL-60 Cells.

Boly R, Franck T, Kohnen S, Lompo M, Guissou IP, Dubois J, Serteyn D, Mouithys-Mickalad A - Evid Based Complement Alternat Med (2015)

UPLC-UV chromatogram profile of the ethyl acetate subfraction acquired at 280 nm with the photodiode array detector. The subfraction was solubilized in methanol and injected onto an Acquity UPLC BEH Shield RP18 column (2.1 × 100 mm, 1.7 μm; Waters). Peaks 1: gallic acid; 2: catechin; 3: unknown; 4: unknown.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4363892&req=5

fig3: UPLC-UV chromatogram profile of the ethyl acetate subfraction acquired at 280 nm with the photodiode array detector. The subfraction was solubilized in methanol and injected onto an Acquity UPLC BEH Shield RP18 column (2.1 × 100 mm, 1.7 μm; Waters). Peaks 1: gallic acid; 2: catechin; 3: unknown; 4: unknown.
Mentions: The lowest IC50 value was found for the butanolic subfraction (IC50 = 0.3 ± 0.1 μg/mL) but was statistically similar to that obtained with the ethyl acetate subfraction (IC50 = 0.4 ± 0.2 μg/mL) and gallic acid (IC50 = 0.4 ± 0.1 μg/mL). Lucigenin is a well-known luminescent probe that is widely used for its high specificity towards superoxide radical [26]. Our results strongly suggest that Agelanthus dodoneifolius subfractions are potential scavengers of superoxide radical, arising from the NADPH oxidase activation, and so that constitutes the first step in the generation of most of the other ROS [27]. Polyphenols such as flavonoids are known to inhibit enzymes responsible for superoxide anion production such as xanthine oxidase, protein kinase C, and NADPH oxidase [28]. It has been suggested that the health benefits afforded by the consumption of fruits, legumes, vegetables, and whole grains are mainly due to the presence of natural antioxidants such as ascorbic acid, carotenoids, tocopherols, and phenolic compounds which represent the most important class [25, 28, 29]. The significant effect of the ethyl acetate and n-butanol subfractions is related to their content of polyphenols compounds as demonstrated by the UPLC-MS/MS analysis (Figure 3).

Bottom Line: The results showed the potent activity of the butanolic subfraction, at least in respect of the chemiluminescence test (IC50 = 0.3 ± 0.1 µg/mL) and the ELISA and SIEFED assays (IC50 = 2.8 ± 1.2 µg/mL and 1.3 ± 1.0 µg/mL), respectively.However, the ethyl acetate subfraction was found to be the most potent in the DCF assay as at the highest concentration, DCF fluorescence intensity decreases of about 50%.Nevertheless, taken together, our results provide evidence that Agelanthus dodoneifolius subfractions may represent potential sources of natural antioxidants and of antimyeloperoxidase compounds.

View Article: PubMed Central - PubMed

Affiliation: Institute for Research in Health Sciences (IRSS/CNRST), 03 BP 7192 Ouagadougou 03, Burkina Faso.

ABSTRACT
The ethyl acetate and n-butanolic subfractions of Agelanthus dodoneifolius were investigated for their antioxidant and antimyeloperoxidase (MPO) activities. The reactive oxygen species (ROS) generation was assessed by lucigenin-enhanced chemiluminescence (CL) and dichlorofluorescein- (DCF-) induced fluorescence techniques from phorbol myristate acetate- (PMA-) stimulated equine neutrophils and human myeloid cell line HL-60, respectively. In parallel, the effects of the tested subfractions were evaluated on the total MPO release by stimulated neutrophils and on the specific MPO activity by means of immunological assays. The results showed the potent activity of the butanolic subfraction, at least in respect of the chemiluminescence test (IC50 = 0.3 ± 0.1 µg/mL) and the ELISA and SIEFED assays (IC50 = 2.8 ± 1.2 µg/mL and 1.3 ± 1.0 µg/mL), respectively. However, the ethyl acetate subfraction was found to be the most potent in the DCF assay as at the highest concentration, DCF fluorescence intensity decreases of about 50%. Moreover, we demonstrated that the ethyl acetate subfraction was rich in catechin (16.51%) while it was not easy to identify the main compounds in the butanolic subfraction using the UPLC-MS/MS technique. Nevertheless, taken together, our results provide evidence that Agelanthus dodoneifolius subfractions may represent potential sources of natural antioxidants and of antimyeloperoxidase compounds.

No MeSH data available.


Related in: MedlinePlus