Anti-inflammatory activity of flavonoid rich fractions of Nectandra angustifolia ethanolic extract

Autores
Ferrini, Leandro Adrian; Ojeda, Gonzalo Adrian; Torres, Ana María; Aguirre, María Victoria; Hernández Garrido, Marita; Nieto Callejo, María Luisa; Rodríguez, Juan Pablo
Año de publicación
2024
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
We have previously shown that the ethanolic extract of Nectandra angustifolia (NaE), from its aerial parts (leaves and stems), has notable anti-inflammatory effects in both in vitro and in vivo studies. These effects were demonstrated in LPS-stimulated macrophage cultures (RAW 264.7) and the BV2 microglial cell line. This study further explores NaE’s anti-inflammatory properties, focusing on its mechanisms in microglia using an in vitro approach.NaE was fractionated by flash chromatography and thin layer chromatography (TLC). Selected fractions (F4, F5, F6, F9) were tested for their activity. Concentrations were based on previous work with NaE. Screening involved measuring nitric oxide production (Griess reaction), IL-6, TNFα (ELISAs), and iNOS and COX-2 expression (western blot). No significant differences were observed compared to NaE at 50 μg/mL. Thus, fractions F5 and F6 were further tested at 25 μg/mL, showing improved results over the whole extract, with F5 selected for deeper analysis.Fraction F5 inhibited p65 subunit translocation to the nucleus, as seen by confocal microscopy, and reduced phosphorylation of p65 and IκBα. Western blot analysis also showed decreased phosphorylation of c-JUN compared to LPS. This suggests an upstream inhibition mechanism. Inhibitors of laminin receptor 67 and TGR5 were also tested but did not affect the observed bioactivity.We examined IL-4 polarization effects, finding that F5 did not induce arginase-1 expression nor alter it after IL-4 stimulation. Additionally, we investigated flavonoid localization within treated cells. Staining showed flavonoids present inside cells, with increased fluorescence after 24 hours.In summary, our study highlights the anti-inflammatory activity of a fraction of NaE. The localization of flavonoids and further mechanistic insights will guide future research on their interactions with upstream targets in LPS-triggered inflammatory pathways.
Fil: Ferrini, Leandro Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
Fil: Ojeda, Gonzalo Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
Fil: Torres, Ana María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
Fil: Aguirre, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
Fil: Hernández Garrido, Marita. Universidad de Valladolid; España
Fil: Nieto Callejo, María Luisa. Universidad de Valladolid; España
Fil: Rodríguez, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
LVI Reunión de la Asociación Argentina de Farmacología Experimental
Bahia Blanca
Argentina
Asociación Argentina de Farmacología Experimental
Universidad Nacional del Sur
Materia
Flavonoid
Natural Product
Inflammation
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/275565
Acceder
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network_name_str CONICET Digital (CONICET)
spelling Anti-inflammatory activity of flavonoid rich fractions of Nectandra angustifolia ethanolic extractFerrini, Leandro AdrianOjeda, Gonzalo AdrianTorres, Ana MaríaAguirre, María VictoriaHernández Garrido, MaritaNieto Callejo, María LuisaRodríguez, Juan PabloFlavonoidNatural ProductInflammationhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1We have previously shown that the ethanolic extract of Nectandra angustifolia (NaE), from its aerial parts (leaves and stems), has notable anti-inflammatory effects in both in vitro and in vivo studies. These effects were demonstrated in LPS-stimulated macrophage cultures (RAW 264.7) and the BV2 microglial cell line. This study further explores NaE’s anti-inflammatory properties, focusing on its mechanisms in microglia using an in vitro approach.NaE was fractionated by flash chromatography and thin layer chromatography (TLC). Selected fractions (F4, F5, F6, F9) were tested for their activity. Concentrations were based on previous work with NaE. Screening involved measuring nitric oxide production (Griess reaction), IL-6, TNFα (ELISAs), and iNOS and COX-2 expression (western blot). No significant differences were observed compared to NaE at 50 μg/mL. Thus, fractions F5 and F6 were further tested at 25 μg/mL, showing improved results over the whole extract, with F5 selected for deeper analysis.Fraction F5 inhibited p65 subunit translocation to the nucleus, as seen by confocal microscopy, and reduced phosphorylation of p65 and IκBα. Western blot analysis also showed decreased phosphorylation of c-JUN compared to LPS. This suggests an upstream inhibition mechanism. Inhibitors of laminin receptor 67 and TGR5 were also tested but did not affect the observed bioactivity.We examined IL-4 polarization effects, finding that F5 did not induce arginase-1 expression nor alter it after IL-4 stimulation. Additionally, we investigated flavonoid localization within treated cells. Staining showed flavonoids present inside cells, with increased fluorescence after 24 hours.In summary, our study highlights the anti-inflammatory activity of a fraction of NaE. The localization of flavonoids and further mechanistic insights will guide future research on their interactions with upstream targets in LPS-triggered inflammatory pathways.Fil: Ferrini, Leandro Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Ojeda, Gonzalo Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Torres, Ana María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Aguirre, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Hernández Garrido, Marita. Universidad de Valladolid; EspañaFil: Nieto Callejo, María Luisa. Universidad de Valladolid; EspañaFil: Rodríguez, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaLVI Reunión de la Asociación Argentina de Farmacología ExperimentalBahia BlancaArgentinaAsociación Argentina de Farmacología ExperimentalUniversidad Nacional del SurAsociación Argentina de Farmacología Experimental2024info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/275565Anti-inflammatory activity of flavonoid rich fractions of Nectandra angustifolia ethanolic extract; LVI Reunión de la Asociación Argentina de Farmacología Experimental; Bahia Blanca; Argentina; 2024; 27-27978-631-90806-0-5CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://aafeargentina.org/congresos-aafe/Internacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-12-03T09:27:58Zoai:ri.conicet.gov.ar:11336/275565instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-12-03 09:27:58.38CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Anti-inflammatory activity of flavonoid rich fractions of Nectandra angustifolia ethanolic extract
title Anti-inflammatory activity of flavonoid rich fractions of Nectandra angustifolia ethanolic extract
spellingShingle Anti-inflammatory activity of flavonoid rich fractions of Nectandra angustifolia ethanolic extract
Ferrini, Leandro Adrian
Flavonoid
Natural Product
Inflammation
title_short Anti-inflammatory activity of flavonoid rich fractions of Nectandra angustifolia ethanolic extract
title_full Anti-inflammatory activity of flavonoid rich fractions of Nectandra angustifolia ethanolic extract
title_fullStr Anti-inflammatory activity of flavonoid rich fractions of Nectandra angustifolia ethanolic extract
title_full_unstemmed Anti-inflammatory activity of flavonoid rich fractions of Nectandra angustifolia ethanolic extract
title_sort Anti-inflammatory activity of flavonoid rich fractions of Nectandra angustifolia ethanolic extract
dc.creator.none.fl_str_mv Ferrini, Leandro Adrian
Ojeda, Gonzalo Adrian
Torres, Ana María
Aguirre, María Victoria
Hernández Garrido, Marita
Nieto Callejo, María Luisa
Rodríguez, Juan Pablo
author Ferrini, Leandro Adrian
author_facet Ferrini, Leandro Adrian
Ojeda, Gonzalo Adrian
Torres, Ana María
Aguirre, María Victoria
Hernández Garrido, Marita
Nieto Callejo, María Luisa
Rodríguez, Juan Pablo
author_role author
author2 Ojeda, Gonzalo Adrian
Torres, Ana María
Aguirre, María Victoria
Hernández Garrido, Marita
Nieto Callejo, María Luisa
Rodríguez, Juan Pablo
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Flavonoid
Natural Product
Inflammation
topic Flavonoid
Natural Product
Inflammation
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv We have previously shown that the ethanolic extract of Nectandra angustifolia (NaE), from its aerial parts (leaves and stems), has notable anti-inflammatory effects in both in vitro and in vivo studies. These effects were demonstrated in LPS-stimulated macrophage cultures (RAW 264.7) and the BV2 microglial cell line. This study further explores NaE’s anti-inflammatory properties, focusing on its mechanisms in microglia using an in vitro approach.NaE was fractionated by flash chromatography and thin layer chromatography (TLC). Selected fractions (F4, F5, F6, F9) were tested for their activity. Concentrations were based on previous work with NaE. Screening involved measuring nitric oxide production (Griess reaction), IL-6, TNFα (ELISAs), and iNOS and COX-2 expression (western blot). No significant differences were observed compared to NaE at 50 μg/mL. Thus, fractions F5 and F6 were further tested at 25 μg/mL, showing improved results over the whole extract, with F5 selected for deeper analysis.Fraction F5 inhibited p65 subunit translocation to the nucleus, as seen by confocal microscopy, and reduced phosphorylation of p65 and IκBα. Western blot analysis also showed decreased phosphorylation of c-JUN compared to LPS. This suggests an upstream inhibition mechanism. Inhibitors of laminin receptor 67 and TGR5 were also tested but did not affect the observed bioactivity.We examined IL-4 polarization effects, finding that F5 did not induce arginase-1 expression nor alter it after IL-4 stimulation. Additionally, we investigated flavonoid localization within treated cells. Staining showed flavonoids present inside cells, with increased fluorescence after 24 hours.In summary, our study highlights the anti-inflammatory activity of a fraction of NaE. The localization of flavonoids and further mechanistic insights will guide future research on their interactions with upstream targets in LPS-triggered inflammatory pathways.
Fil: Ferrini, Leandro Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
Fil: Ojeda, Gonzalo Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
Fil: Torres, Ana María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
Fil: Aguirre, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
Fil: Hernández Garrido, Marita. Universidad de Valladolid; España
Fil: Nieto Callejo, María Luisa. Universidad de Valladolid; España
Fil: Rodríguez, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
LVI Reunión de la Asociación Argentina de Farmacología Experimental
Bahia Blanca
Argentina
Asociación Argentina de Farmacología Experimental
Universidad Nacional del Sur
description We have previously shown that the ethanolic extract of Nectandra angustifolia (NaE), from its aerial parts (leaves and stems), has notable anti-inflammatory effects in both in vitro and in vivo studies. These effects were demonstrated in LPS-stimulated macrophage cultures (RAW 264.7) and the BV2 microglial cell line. This study further explores NaE’s anti-inflammatory properties, focusing on its mechanisms in microglia using an in vitro approach.NaE was fractionated by flash chromatography and thin layer chromatography (TLC). Selected fractions (F4, F5, F6, F9) were tested for their activity. Concentrations were based on previous work with NaE. Screening involved measuring nitric oxide production (Griess reaction), IL-6, TNFα (ELISAs), and iNOS and COX-2 expression (western blot). No significant differences were observed compared to NaE at 50 μg/mL. Thus, fractions F5 and F6 were further tested at 25 μg/mL, showing improved results over the whole extract, with F5 selected for deeper analysis.Fraction F5 inhibited p65 subunit translocation to the nucleus, as seen by confocal microscopy, and reduced phosphorylation of p65 and IκBα. Western blot analysis also showed decreased phosphorylation of c-JUN compared to LPS. This suggests an upstream inhibition mechanism. Inhibitors of laminin receptor 67 and TGR5 were also tested but did not affect the observed bioactivity.We examined IL-4 polarization effects, finding that F5 did not induce arginase-1 expression nor alter it after IL-4 stimulation. Additionally, we investigated flavonoid localization within treated cells. Staining showed flavonoids present inside cells, with increased fluorescence after 24 hours.In summary, our study highlights the anti-inflammatory activity of a fraction of NaE. The localization of flavonoids and further mechanistic insights will guide future research on their interactions with upstream targets in LPS-triggered inflammatory pathways.
publishDate 2024
dc.date.none.fl_str_mv 2024
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Book
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/275565
Anti-inflammatory activity of flavonoid rich fractions of Nectandra angustifolia ethanolic extract; LVI Reunión de la Asociación Argentina de Farmacología Experimental; Bahia Blanca; Argentina; 2024; 27-27
978-631-90806-0-5
CONICET Digital
CONICET
url http://hdl.handle.net/11336/275565
identifier_str_mv Anti-inflammatory activity of flavonoid rich fractions of Nectandra angustifolia ethanolic extract; LVI Reunión de la Asociación Argentina de Farmacología Experimental; Bahia Blanca; Argentina; 2024; 27-27
978-631-90806-0-5
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://aafeargentina.org/congresos-aafe/
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eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.coverage.none.fl_str_mv Internacional
dc.publisher.none.fl_str_mv Asociación Argentina de Farmacología Experimental
publisher.none.fl_str_mv Asociación Argentina de Farmacología Experimental
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
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repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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