<i>Allophylus africanus</i> Stem Bark Extract Modulates the Mitochondrial Apoptotic Pathway in Human Stomach Cancer Cells

The present work aimed to detail the mechanisms elicited by <i>Allophylus africanus</i> P. Beauv. stem bark extract in human stomach cancer cells and to identify the bioactives underlying the cytotoxicity. MTT reduction and LDH leakage assays allowed characterizing the cytotoxic effects...

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Huvudupphovsmän: Vera Ribeiro, Federico Ferreres, Andreia Oliveira, Nelson G. M. Gomes, Ángel Gil-Izquierdo, Luísa Araújo, David Pereira, Patrícia Valentão
Materialtyp: Artikel
Språk:English
Publicerad: MDPI AG 2023-02-01
Serie:Life
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Länkar:https://www.mdpi.com/2075-1729/13/2/406
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author Vera Ribeiro
Federico Ferreres
Andreia Oliveira
Nelson G. M. Gomes
Ángel Gil-Izquierdo
Luísa Araújo
David Pereira
Patrícia Valentão
author_facet Vera Ribeiro
Federico Ferreres
Andreia Oliveira
Nelson G. M. Gomes
Ángel Gil-Izquierdo
Luísa Araújo
David Pereira
Patrícia Valentão
author_sort Vera Ribeiro
collection DOAJ
description The present work aimed to detail the mechanisms elicited by <i>Allophylus africanus</i> P. Beauv. stem bark extract in human stomach cancer cells and to identify the bioactives underlying the cytotoxicity. MTT reduction and LDH leakage assays allowed characterizing the cytotoxic effects in AGS cells, which were further detailed by morphological analysis using phalloidin and Hoechst 33258. Proapoptotic mechanisms were elucidated through a mitochondrial membrane potential assay and by assessing the impact upon the activity of caspase-9 and -3. The extract displayed selective cytotoxicity against AGS cells. The absence of plasma membrane permeabilization, along with apoptotic body formation, suggested that pro-apoptotic effects triggered cell death. Intrinsic apoptosis pathway activation was verified, as mitochondrial membrane potential decrease and activation of caspase-9 and -3 were observed. HPLC-DAD profiling enabled the identification of two apigenin-di-<i>C</i>-glycosides, vicenin-2 (<b>1</b>) and apigenin-6<i>-C-</i>hexoside-8<i>-C-</i>pentoside (<b>3</b>), as well as three mono<i>-C-</i>glycosides-<i>O</i>-glycosylated derivatives, apigenin-7<i>-O-</i>hexoside-8<i>-C-</i>hexoside (<b>2</b>), apigenin-8<i>-C-</i>(2-rhamnosyl)hexoside (<b>4</b>) and apigenin-6<i>-C-</i>(2-rhamnosyl)hexoside (<b>5</b>). Isovitexin-2″<i>-O-</i>rhamnoside (<b>5</b>) is the main constituent, accounting for nearly 40% of the total quantifiable flavonoid content. Our results allowed us to establish the relationship between the presence of vicenin-2 and other apigenin derivatives with the contribution to the cytotoxic effects on the presented AGS cells. Our findings attest the anticancer potential of <i>A. africanus</i> stem bark against gastric adenocarcinoma, calling for studies to develop herbal-based products and/or the use of apigenin derivatives in chemotherapeutic drug development.
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spelling doaj.art-1d55d85b89dd45d8856d946b17e45d282023-11-16T21:40:50ZengMDPI AGLife2075-17292023-02-0113240610.3390/life13020406<i>Allophylus africanus</i> Stem Bark Extract Modulates the Mitochondrial Apoptotic Pathway in Human Stomach Cancer CellsVera Ribeiro0Federico Ferreres1Andreia Oliveira2Nelson G. M. Gomes3Ángel Gil-Izquierdo4Luísa Araújo5David Pereira6Patrícia Valentão7REQUIMTE/LAQV, Laboratório de Farmacognosia, Departamento de Química, Faculdade de Farmácia, Universidade do Porto, R. Jorge Viterbo Ferreira, nº 228, 4050-313 Porto, PortugalDepartment of Food Technology and Nutrition, Molecular Recognition and Encapsulation (REM) Group, Universidad Católica de Murcia, UCAM, Campus Los Jerónimos, s/n, 30107 Murcia, SpainREQUIMTE/LAQV, Laboratório de Farmacognosia, Departamento de Química, Faculdade de Farmácia, Universidade do Porto, R. Jorge Viterbo Ferreira, nº 228, 4050-313 Porto, PortugalREQUIMTE/LAQV, Laboratório de Farmacognosia, Departamento de Química, Faculdade de Farmácia, Universidade do Porto, R. Jorge Viterbo Ferreira, nº 228, 4050-313 Porto, PortugalResearch Group on Quality, Safety and Bioactivity of Plant Foods, Department of Food Science and Technology, CEBAS (CSIC), Campus University Espinardo, P.O. Box 164, 30100 Murcia, SpainMDS—Medicamentos e Diagnósticos em Saúde, Avenida dos Combatentes da Liberdade da Pátria, Bissau, Guinea-BissauREQUIMTE/LAQV, Laboratório de Farmacognosia, Departamento de Química, Faculdade de Farmácia, Universidade do Porto, R. Jorge Viterbo Ferreira, nº 228, 4050-313 Porto, PortugalREQUIMTE/LAQV, Laboratório de Farmacognosia, Departamento de Química, Faculdade de Farmácia, Universidade do Porto, R. Jorge Viterbo Ferreira, nº 228, 4050-313 Porto, PortugalThe present work aimed to detail the mechanisms elicited by <i>Allophylus africanus</i> P. Beauv. stem bark extract in human stomach cancer cells and to identify the bioactives underlying the cytotoxicity. MTT reduction and LDH leakage assays allowed characterizing the cytotoxic effects in AGS cells, which were further detailed by morphological analysis using phalloidin and Hoechst 33258. Proapoptotic mechanisms were elucidated through a mitochondrial membrane potential assay and by assessing the impact upon the activity of caspase-9 and -3. The extract displayed selective cytotoxicity against AGS cells. The absence of plasma membrane permeabilization, along with apoptotic body formation, suggested that pro-apoptotic effects triggered cell death. Intrinsic apoptosis pathway activation was verified, as mitochondrial membrane potential decrease and activation of caspase-9 and -3 were observed. HPLC-DAD profiling enabled the identification of two apigenin-di-<i>C</i>-glycosides, vicenin-2 (<b>1</b>) and apigenin-6<i>-C-</i>hexoside-8<i>-C-</i>pentoside (<b>3</b>), as well as three mono<i>-C-</i>glycosides-<i>O</i>-glycosylated derivatives, apigenin-7<i>-O-</i>hexoside-8<i>-C-</i>hexoside (<b>2</b>), apigenin-8<i>-C-</i>(2-rhamnosyl)hexoside (<b>4</b>) and apigenin-6<i>-C-</i>(2-rhamnosyl)hexoside (<b>5</b>). Isovitexin-2″<i>-O-</i>rhamnoside (<b>5</b>) is the main constituent, accounting for nearly 40% of the total quantifiable flavonoid content. Our results allowed us to establish the relationship between the presence of vicenin-2 and other apigenin derivatives with the contribution to the cytotoxic effects on the presented AGS cells. Our findings attest the anticancer potential of <i>A. africanus</i> stem bark against gastric adenocarcinoma, calling for studies to develop herbal-based products and/or the use of apigenin derivatives in chemotherapeutic drug development.https://www.mdpi.com/2075-1729/13/2/406<i>Allophylus africanus</i>EthnopharmacologyisovitexinRhamnosylvitexinSapindaceaevitexin
spellingShingle Vera Ribeiro
Federico Ferreres
Andreia Oliveira
Nelson G. M. Gomes
Ángel Gil-Izquierdo
Luísa Araújo
David Pereira
Patrícia Valentão
<i>Allophylus africanus</i> Stem Bark Extract Modulates the Mitochondrial Apoptotic Pathway in Human Stomach Cancer Cells
Life
<i>Allophylus africanus</i>
Ethnopharmacology
isovitexin
Rhamnosylvitexin
Sapindaceae
vitexin
title <i>Allophylus africanus</i> Stem Bark Extract Modulates the Mitochondrial Apoptotic Pathway in Human Stomach Cancer Cells
title_full <i>Allophylus africanus</i> Stem Bark Extract Modulates the Mitochondrial Apoptotic Pathway in Human Stomach Cancer Cells
title_fullStr <i>Allophylus africanus</i> Stem Bark Extract Modulates the Mitochondrial Apoptotic Pathway in Human Stomach Cancer Cells
title_full_unstemmed <i>Allophylus africanus</i> Stem Bark Extract Modulates the Mitochondrial Apoptotic Pathway in Human Stomach Cancer Cells
title_short <i>Allophylus africanus</i> Stem Bark Extract Modulates the Mitochondrial Apoptotic Pathway in Human Stomach Cancer Cells
title_sort i allophylus africanus i stem bark extract modulates the mitochondrial apoptotic pathway in human stomach cancer cells
topic <i>Allophylus africanus</i>
Ethnopharmacology
isovitexin
Rhamnosylvitexin
Sapindaceae
vitexin
url https://www.mdpi.com/2075-1729/13/2/406
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