Antioxidant Effects of Protocatechuic Acid, Ferulic Acid, and Caffeic Acid in Human Neutrophils Using a Fluorescent Substance

Human neutrophils stimulated by phorbol myristate acetate (PMA), an activator of protein kinase C, produce active oxygen by NADPH oxidase in intracellular structures. We added succinimidyl ester of dichlorodihydrofluorescein diacetate (H2DCFDA), which first emits fluorescence when oxidized with acti...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Hyogo,Akiyo, Kobayashi,Toshihiro, Garcia del Saz,Eva, Seguchi,Harumichi
Lenguaje:	English
Publicado:	Sociedad Chilena de Anatomía 2010
Materias:	Antioxidant effects Polyphenols Human neutrophils Flow cytometry
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022010000300040
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:scielo:S0717-95022010000300040
record_format	dspace
spelling	oai:scielo:S0717-950220100003000402010-10-25Antioxidant Effects of Protocatechuic Acid, Ferulic Acid, and Caffeic Acid in Human Neutrophils Using a Fluorescent SubstanceHyogo,AkiyoKobayashi,ToshihiroGarcia del Saz,EvaSeguchi,Harumichi Antioxidant effects Polyphenols Human neutrophils Flow cytometry Human neutrophils stimulated by phorbol myristate acetate (PMA), an activator of protein kinase C, produce active oxygen by NADPH oxidase in intracellular structures. We added succinimidyl ester of dichlorodihydrofluorescein diacetate (H2DCFDA), which first emits fluorescence when oxidized with active oxygen species, to neutrophils to produce active oxygen, in order to investigate the antioxidant effects of protocatechuic acid, ferulic acid, and caffeic acid which belong to polyphenols and are widely distributed among plants. Particularly, we focused on examining whether these substances capture and eliminate active oxygen inside or outside the neutrophil cytoplasm and whether these substances inhibit NADPH oxidase. Fluorescence microscopy demonstrated that fluorescence-positive intracellular structures were decreased in neutrophils when stimulated by PMA and exposed to an antioxidant. Quantitative measurement by flow cytometry revealed that the fluorescence intensities in neutrophils, exposed to protocatechuic acid, ferulic acid, or caffeic acid, were decreased by 62.9%, 71.4%, and 86.1%, respectively, as compared with those stimulated by PMA but not exposed to an antioxidant. Judging from fluorescence microscopy and dot blots by flow cytometry, these antioxidants had no effects on neutrophil morphology. On the other hand, the fluorescence intensities of the active oxygen released from neutrophils were decreased by 81.4%, 46.7%, and 27.4%, respectively. Diphenylene iodonium, a specific inhibitor of NADPH oxidase, inhibited the enzyme by 92.1% in the PMA-stimulated neutrophils. Protocatechuic acid, ferulic acid, and caffeic acid inhibited the enzyme by 36.5%, 54.6%, and 27.4%, respectively. These results demonstrate that protocatechuic acid, ferulic acid, and caffeic acid capture and eliminate active oxygen, produced by PMA-stimulated neutrophils, intracellularly and extracellularly. Furthermore, these antioxidants partially inhibit NADPH oxidase activity.info:eu-repo/semantics/openAccessSociedad Chilena de AnatomíaInternational Journal of Morphology v.28 n.3 20102010-09-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022010000300040en10.4067/S0717-95022010000300040
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Antioxidant effects Polyphenols Human neutrophils Flow cytometry
spellingShingle	Antioxidant effects Polyphenols Human neutrophils Flow cytometry Hyogo,Akiyo Kobayashi,Toshihiro Garcia del Saz,Eva Seguchi,Harumichi Antioxidant Effects of Protocatechuic Acid, Ferulic Acid, and Caffeic Acid in Human Neutrophils Using a Fluorescent Substance
description	Human neutrophils stimulated by phorbol myristate acetate (PMA), an activator of protein kinase C, produce active oxygen by NADPH oxidase in intracellular structures. We added succinimidyl ester of dichlorodihydrofluorescein diacetate (H2DCFDA), which first emits fluorescence when oxidized with active oxygen species, to neutrophils to produce active oxygen, in order to investigate the antioxidant effects of protocatechuic acid, ferulic acid, and caffeic acid which belong to polyphenols and are widely distributed among plants. Particularly, we focused on examining whether these substances capture and eliminate active oxygen inside or outside the neutrophil cytoplasm and whether these substances inhibit NADPH oxidase. Fluorescence microscopy demonstrated that fluorescence-positive intracellular structures were decreased in neutrophils when stimulated by PMA and exposed to an antioxidant. Quantitative measurement by flow cytometry revealed that the fluorescence intensities in neutrophils, exposed to protocatechuic acid, ferulic acid, or caffeic acid, were decreased by 62.9%, 71.4%, and 86.1%, respectively, as compared with those stimulated by PMA but not exposed to an antioxidant. Judging from fluorescence microscopy and dot blots by flow cytometry, these antioxidants had no effects on neutrophil morphology. On the other hand, the fluorescence intensities of the active oxygen released from neutrophils were decreased by 81.4%, 46.7%, and 27.4%, respectively. Diphenylene iodonium, a specific inhibitor of NADPH oxidase, inhibited the enzyme by 92.1% in the PMA-stimulated neutrophils. Protocatechuic acid, ferulic acid, and caffeic acid inhibited the enzyme by 36.5%, 54.6%, and 27.4%, respectively. These results demonstrate that protocatechuic acid, ferulic acid, and caffeic acid capture and eliminate active oxygen, produced by PMA-stimulated neutrophils, intracellularly and extracellularly. Furthermore, these antioxidants partially inhibit NADPH oxidase activity.
author	Hyogo,Akiyo Kobayashi,Toshihiro Garcia del Saz,Eva Seguchi,Harumichi
author_facet	Hyogo,Akiyo Kobayashi,Toshihiro Garcia del Saz,Eva Seguchi,Harumichi
author_sort	Hyogo,Akiyo
title	Antioxidant Effects of Protocatechuic Acid, Ferulic Acid, and Caffeic Acid in Human Neutrophils Using a Fluorescent Substance
title_short	Antioxidant Effects of Protocatechuic Acid, Ferulic Acid, and Caffeic Acid in Human Neutrophils Using a Fluorescent Substance
title_full	Antioxidant Effects of Protocatechuic Acid, Ferulic Acid, and Caffeic Acid in Human Neutrophils Using a Fluorescent Substance
title_fullStr	Antioxidant Effects of Protocatechuic Acid, Ferulic Acid, and Caffeic Acid in Human Neutrophils Using a Fluorescent Substance
title_full_unstemmed	Antioxidant Effects of Protocatechuic Acid, Ferulic Acid, and Caffeic Acid in Human Neutrophils Using a Fluorescent Substance
title_sort	antioxidant effects of protocatechuic acid, ferulic acid, and caffeic acid in human neutrophils using a fluorescent substance
publisher	Sociedad Chilena de Anatomía
publishDate	2010
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022010000300040
work_keys_str_mv	AT hyogoakiyo antioxidanteffectsofprotocatechuicacidferulicacidandcaffeicacidinhumanneutrophilsusingafluorescentsubstance AT kobayashitoshihiro antioxidanteffectsofprotocatechuicacidferulicacidandcaffeicacidinhumanneutrophilsusingafluorescentsubstance AT garciadelsazeva antioxidanteffectsofprotocatechuicacidferulicacidandcaffeicacidinhumanneutrophilsusingafluorescentsubstance AT seguchiharumichi antioxidanteffectsofprotocatechuicacidferulicacidandcaffeicacidinhumanneutrophilsusingafluorescentsubstance
_version_	1718444705864220672

Antioxidant Effects of Protocatechuic Acid, Ferulic Acid, and Caffeic Acid in Human Neutrophils Using a Fluorescent Substance

Ejemplares similares