Leaf and Root Extracts from Campomanesia adamantium (Myrtaceae) Promote Apoptotic Death of Leukemic Cells via Activation of Intracellular Calcium and Caspase-3

Phytochemical studies are seeking new alternatives to prevent or treat cancer, including different types of leukemias. Campomanesia adamantium, commonly known as guavira or guabiroba, exhibits pharmacological properties including antioxidant, antimicrobial, and antiproliferative activities. Considering the anticancer potential of this plant species, the aim of this study was to evaluate the antileukemic activity and the chemical composition of aqueous extracts from the leaves (AECL) and roots (AECR) of C. adamantium and their possible mechanisms of action. The extracts were analyzed by LC-DAD-MS, and their constituents were identified based on the UV, MS, and MS/MS data. The AECL and AECR showed different chemical compositions, which were identified as main compounds glycosylated flavonols from AECL and ellagic acid and their derivatives from AECR. The cytotoxicity promoted by these extracts were evaluated using human peripheral blood mononuclear cells and Jurkat leukemic cell line. The cell death profile was evaluated using annexin-V-FITC and propidium iodide labeling. Changes in the mitochondrial membrane potential, the activity of caspases, and intracellular calcium levels were assessed. The cell cycle profile was evaluated using propidium iodide. Both extracts caused concentration-dependent cytotoxicity only in Jurkat cells via late apoptosis. This activity was associated with loss of the mitochondrial membrane potential, activation of caspases-9 and -3, changes in intracellular calcium levels, and cell cycle arrest in S-phase. Therefore, the antileukemic activity of the AECL and AECR is mediated by mitochondrial dysfunction and intracellular messengers, which activate the intrinsic apoptotic pathway. Hence, aqueous extracts of the leaves and roots of C. adamantium show therapeutic potential for use in the prevention and treatment of diseases associated the proliferation of tumor cell.Fundação de Apoio ao Desenvolvimento do Ensino, Ciência e Tecnologia do Mato Grosso do Sul (FUNDECT)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Instituto Nacional de Pesquisa do Pantanal - INPPFundação de Amparo e Desenvolvimento da Pesquisa FadespFed Univ Grande Dourados, Res Grp Biotechnol & Bioprospecting Appl Metab, Dourados, BrazilUniv Fed São Paulo, Dept Biochem, São Paulo, BrazilUniv Braz Cubas, Fac Pharm, Mogi Das Cruzes, BrazilUniv Mogi das Cruzes, Interdisciplinary Ctr Biochem Invest, Mogi Das Cruzes, BrazilUniv Fed São Paulo, Dept Pharmaceut Sci, São Paulo, BrazilUniv Fed Mato Grosso do Sul, Lab Nat Prod & Mass Spectrometry, Campo Grande, MS, BrazilUniv Fed São Paulo, Dept Biochem, São Paulo, BrazilUniv Fed São Paulo, Dept Pharmaceut Sci, São Paulo, BrazilWeb of Scienc

Esferas de quitosana/Fe na degradação do corante Azul QR-19 por processos foto-Fenton utilizando luz artificial ou solar Chitosan/Fe spheres on the Blue QR-19 dye degradation by photo Fenton processes using artificial or solar light

A contaminação dos recursos hídricos é um dos maiores problemas ambientais da atualidade. Dentre as várias fontes poluidoras, destacam-se as indústrias têxteis, por serem fontes geradoras de grandes volumes de efluentes, muitas vezes tratados de maneira ineficiente. A principal causa do grande impacto ambiental decorrente deste descarte é a presença dos corantes, tais como os compostos do tipo azo, que podem gerar subprodutos de caráter carcinogênico e/ou mutagênico, ou como os de base antraquinona, que são muito resistentes à degradação natural e portanto persistem no efluente por um longo tempo. O objetivo do presente trabalho consiste na utilização de processos foto-Fenton assistidos por luz artificial e solar, utilizando ferro imobilizado em esferas de quitosana reticulada com glutaraldeído, para promover a degradação do corante reativo Azul QR-19, de base antraquinona, em solução aquosa. As esferas obtidas apresentaram tamanho regular com diâmetro de 4,0 mm. Os resultados demonstraram 90% de descoloração do sistema em 180minutos e redução de 60% do teor de carbono orgânico total (COT), para o sistema foto-Fenton utilizando luz artificial. Para o sistema fotoassistido com luz solar, a descoloração foi integralmente obtida em 120 minutos com 70% de redução do teor de COT. Foi observado que o ferro permaneceu na matriz após o tratamento, possibilitando sua reutilização.<br>The contamination of water resources is one of the greatest environmental problems today. Among the polluting sources are the textile industries due to the production of large volumes of effluent, often treated inefficiently. The main reason for the environmental impact of the rejected materials is the presence of dyes such as azo reactive compounds that can generate by-products with carcinogenic and mutagenic effects. They may also include anthraquinone type compounds, which are highly resistant to degradation and persist in the effluent for a long time. The purpose of this study is the use of photo-Fenton processes assisted by artificial or solar light, using immobilized iron on chitosan beads, crosslinked with glutaraldheyde, for the antraquinone type compound Blue QR-19 standard dye degradation in aqueous solutions. The obtained spheres showed a regular size and 4.0 mm diameter. The results showed 90% discolouration of the system within 180 minutes and a 60% total organic carbon (TOC) reduction for the photo-Fenton system using artificial light. For the system using sunlight, the total discolouration was achieved in 120 minutes and the TOC value decreased 70%. Also observed was that iron remained in the polymeric matrix after the treatment, thus allowing reuse

Chemical composition.

<p>Chromatograms obtained by HPLC-DAD-MS/MS (negative ion mode) indicating the presence of phenolic compounds in the EEHS. The enlarged figures correspond to the MS/MS spectra of the following main compounds: chlorogenic acid (peak 2) and rutin (peak 6).</p

Cytotoxic activity of the EEHS in the acute myeloid leukemia cell line Kasumi-1 in the presence of cell death inhibitors.

<p>(A) Dot plots of the flow cytometry data of cells stained with propidium iodide and annexin-V-FITC in the presence of inhibitors Z-VAD-FMK, Nec-1, NAC, or E64 and in the presence of the EEHS combined with each of these inhibitors. (B) Percentage of cell viability obtained from dot plots at the EEHS concentration of 160 μg/mL. * P < 0.05 when the treated groups were compared with the control group.</p

Effects of EEHS on the inhibition of hemolysis of human erythrocytes induced by AAPH.

<p>The groups were evaluated at (A) 60, (B) 120, (C) 180, and (D) 240 min of incubation. The controls consisted of an erythrocyte suspension (2.5%) incubated with AAPH alone. Values are expressed as the mean ± SEM of three independent experiments performed in duplicate. * P < 0.05 when the treated groups were compared with the AAPH group (erythrocytes incubated with AAPH alone) during the respective incubation periods.</p

Minimal inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and minimum fungicidal concentration (MFC) of the EEHS and controls.

<p>Minimal inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and minimum fungicidal concentration (MFC) of the EEHS and controls.</p

Effects of the EEHS on the inhibition of lipid peroxidation induced by AAPH.

<p>The control corresponds to erythrocytes incubated with AAPH alone. Values are expressed as the mean ± SEM of two independent experiments performed in duplicate. *** P < 0.001 when the treated groups were compared with the AAPH group (erythrocytes incubated with AAPH alone).</p

Estimation of the effect of the EEHS (160 μg/mL) on the mitochondrial membrane potential of the acute myeloid leukemia cell line Kasumi-1 by flow cytometry.

<p>(A) Dot plots of flow cytometric data indicating the pattern of cell staining using JC-1 in green (FL-1) and red (FL-2). (B) Percentage of change of the mitochondrial membrane potential obtained from dot plots. *** P < 0.001 when the treated groups were compared with the control group.</p

Cytotoxic activity of the EEHS in the acute myeloid leukemia cell line Kasumi-1.

<p>(A) Concentration-response curve (25–200 μg/mL). (B) Dot plots indicating the flow cytometry data of cells stained with propidium iodide (PI) and annexin-V-FITC (An) treated with a negative control (NC), 100 μg/mL of the EEHS or 200 μg/mL of the EEHS. The lower left quadrant shows viable cells (An^–/PI^–); the lower right quadrant shows apoptotic cells (An⁺/PI^–); the upper left quadrant shows cells undergoing necrosis (An^–/PI⁺); and the upper right quadrant shows cells in late apoptosis (An⁺/PI⁺). (C) Percentage of cell death obtained from dot plots of cells treated with NC, 100 μg/mL of the EEHS, or 200 μg/mL of the EEHS. * P < 0.05, ** P < 0.01 and *** P < 0.001 when the treated groups were compared with the control group.</p