Apoptotic activity of isoespintanol and semisynthetic derivatives in human polymorphonuclear cells

Background: Inflammation is a complex physiopathologic response to different stimuli. Recently, some pharmacological strategies have been proposed that could be used for resolution of inflammation by enhancing apoptosis of inflammatory cells. Objectives: To study in vitro apoptotic activity of isoe...

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Autores principales:	Martín DADE, Paula GALEANO, José-Luis RÍOS, Benjamín ROJANO, Guillermo SCHINELLA
Formato:	article
Lenguaje:	EN
Publicado:	Universidad de Antioquia 2016
Materias:	Isoespintanol inflammation neutrophils apoptosis Food processing and manufacture TP368-456 Pharmaceutical industry HD9665-9675
Acceso en línea:	https://doaj.org/article/55c78c362eb0433f8b3dd9cd538b0488
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spelling	oai:doaj.org-article:55c78c362eb0433f8b3dd9cd538b04882021-11-19T04:09:58ZApoptotic activity of isoespintanol and semisynthetic derivatives in human polymorphonuclear cells0121-40042145-2660https://doaj.org/article/55c78c362eb0433f8b3dd9cd538b04882016-07-01T00:00:00Zhttps://revistas.udea.edu.co/index.php/vitae/article/view/22441https://doaj.org/toc/0121-4004https://doaj.org/toc/2145-2660 Background: Inflammation is a complex physiopathologic response to different stimuli. Recently, some pharmacological strategies have been proposed that could be used for resolution of inflammation by enhancing apoptosis of inflammatory cells. Objectives: To study in vitro apoptotic activity of isoespintanol [ISO] and of two semi-synthetic derivatives, bromide isoespintanol [BrI] and demethylated isoespintanol [DMI], in human polymorphonuclear (PMN) cells. Methods: PMN were exposed to the different concentrations of ISO, BrI and DMI for 30 min in phosphate-buffered saline pH 7.4 containing 1 mg/mL glucose, 0.4 mM Mg2+, and 1.20 mM Ca2+. Viability was assessed by dimethylthiazol diphenyl tetrazolium bromide (MTT). To distinguish between the two modes of cell death, apoptosis and necrosis, we examined differences in morphological and biochemical changes of cells stained with annexin V- FITC (An) and/or propidium iodide (PI) using two different assays based on flow cytometry Results: The MTT assay revealed the ability of cells to reduce MTT salt to formazan. In the presence of BrI and DMI a significant concentration-dependent decrease of cell viability was observed. The annexin V- FITC binding assay showed a high proportion of apoptotic cells for those treated with BrI (An+/ PI-: 62.3 ± 8.2% vs. 2.1 ± 0.5% of control, P<0.05). The population of PMN treated with DMI produced the highest percentage (An+/IP+: 43.4 ± 5.2 % vs. 0.4 ± 0.3 % of control, P<0.05) of necrotic cells. Apoptotic nuclei were analyzed by PI staining. The cell population in the sub G0/G1 region represents cells with hypodiploidal DNA, an indicator of apoptosis. When cells were incubated with 50 and 100 μM of BrI, the cell population in the sub G0/G1 region increased, suggesting a dose-dependent increase in the population of apoptotic cells. The presence of the pan-inhibitor of caspases (Z-VAD-fmk) showed a significant reduction in cell population in the sub G0/G1 region, indicating less degradation of DNA. Conclusions: Bromide isoespintanol [BrI] induces an apoptotic process in PMN, mediated –at least in part– by activation of caspases, although this compound may probably act through other caspase-independent mechanisms as well. Martín DADEPaula GALEANOJosé-Luis RÍOSBenjamín ROJANOGuillermo SCHINELLAUniversidad de AntioquiaarticleIsoespintanolinflammationneutrophilsapoptosisFood processing and manufactureTP368-456Pharmaceutical industryHD9665-9675ENVitae, Vol 23, Iss 1 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	Isoespintanol inflammation neutrophils apoptosis Food processing and manufacture TP368-456 Pharmaceutical industry HD9665-9675
spellingShingle	Isoespintanol inflammation neutrophils apoptosis Food processing and manufacture TP368-456 Pharmaceutical industry HD9665-9675 Martín DADE Paula GALEANO José-Luis RÍOS Benjamín ROJANO Guillermo SCHINELLA Apoptotic activity of isoespintanol and semisynthetic derivatives in human polymorphonuclear cells
description	Background: Inflammation is a complex physiopathologic response to different stimuli. Recently, some pharmacological strategies have been proposed that could be used for resolution of inflammation by enhancing apoptosis of inflammatory cells. Objectives: To study in vitro apoptotic activity of isoespintanol [ISO] and of two semi-synthetic derivatives, bromide isoespintanol [BrI] and demethylated isoespintanol [DMI], in human polymorphonuclear (PMN) cells. Methods: PMN were exposed to the different concentrations of ISO, BrI and DMI for 30 min in phosphate-buffered saline pH 7.4 containing 1 mg/mL glucose, 0.4 mM Mg2+, and 1.20 mM Ca2+. Viability was assessed by dimethylthiazol diphenyl tetrazolium bromide (MTT). To distinguish between the two modes of cell death, apoptosis and necrosis, we examined differences in morphological and biochemical changes of cells stained with annexin V- FITC (An) and/or propidium iodide (PI) using two different assays based on flow cytometry Results: The MTT assay revealed the ability of cells to reduce MTT salt to formazan. In the presence of BrI and DMI a significant concentration-dependent decrease of cell viability was observed. The annexin V- FITC binding assay showed a high proportion of apoptotic cells for those treated with BrI (An+/ PI-: 62.3 ± 8.2% vs. 2.1 ± 0.5% of control, P<0.05). The population of PMN treated with DMI produced the highest percentage (An+/IP+: 43.4 ± 5.2 % vs. 0.4 ± 0.3 % of control, P<0.05) of necrotic cells. Apoptotic nuclei were analyzed by PI staining. The cell population in the sub G0/G1 region represents cells with hypodiploidal DNA, an indicator of apoptosis. When cells were incubated with 50 and 100 μM of BrI, the cell population in the sub G0/G1 region increased, suggesting a dose-dependent increase in the population of apoptotic cells. The presence of the pan-inhibitor of caspases (Z-VAD-fmk) showed a significant reduction in cell population in the sub G0/G1 region, indicating less degradation of DNA. Conclusions: Bromide isoespintanol [BrI] induces an apoptotic process in PMN, mediated –at least in part– by activation of caspases, although this compound may probably act through other caspase-independent mechanisms as well.
format	article
author	Martín DADE Paula GALEANO José-Luis RÍOS Benjamín ROJANO Guillermo SCHINELLA
author_facet	Martín DADE Paula GALEANO José-Luis RÍOS Benjamín ROJANO Guillermo SCHINELLA
author_sort	Martín DADE
title	Apoptotic activity of isoespintanol and semisynthetic derivatives in human polymorphonuclear cells
title_short	Apoptotic activity of isoespintanol and semisynthetic derivatives in human polymorphonuclear cells
title_full	Apoptotic activity of isoespintanol and semisynthetic derivatives in human polymorphonuclear cells
title_fullStr	Apoptotic activity of isoespintanol and semisynthetic derivatives in human polymorphonuclear cells
title_full_unstemmed	Apoptotic activity of isoespintanol and semisynthetic derivatives in human polymorphonuclear cells
title_sort	apoptotic activity of isoespintanol and semisynthetic derivatives in human polymorphonuclear cells
publisher	Universidad de Antioquia
publishDate	2016
url	https://doaj.org/article/55c78c362eb0433f8b3dd9cd538b0488
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_version_	1718420495871770624

Apoptotic activity of isoespintanol and semisynthetic derivatives in human polymorphonuclear cells

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