In Vitro Cytotoxic Effects and Mechanisms of Action of Eleutherine Isolated from Eleutherine plicata Bulb in Rat Glioma C6 Cells.

Gliomas are the most common primary malignant brain tumors in adults, and have a poor prognosis, despite the different types of treatment available. There is growing demand for new therapies to treat this life-threatening tumor. Quinone derivatives from plants have received increased interest as potential anti-glioma drugs, due to their diverse pharmacologic activities, such as inhibiting cell growth, inflammation, tumor invasion, and promoting tumor regression. Previous studies have demonstrated the anti-glioma activity of Eleutherine plicata, which is related to three main naphthoquinone compounds-eleutherine, isoeleutherine, and eleutherol-but their mechanism of action remains elusive. Thus, the aim of this study was to investigate the mechanism of action of eleutherine on rat C6 glioma. In vitro cytotoxicity was evaluated by MTT assay; morphological changes were evaluated by phase-contrast microscopy. Apoptosis was determined by annexin V-FITC-propidium iodide staining, and antiproliferative effects were assessed by wound migration and colony formation assays. Protein kinase B (AKT/pAKT) expression was measured by western blot, and telomerase reverse transcriptase mRNA was measured by quantitative real-time polymerase chain reaction (qRT-PCR). Eleutherine reduced C6 cell proliferation in a dose-dependent manner, suppressed migration and invasion, induced apoptosis, and reduced AKT phosphorylation and telomerase expression. In summary, our results suggest that eleutherine has potential clinical use in treating glioma.

Proposta de escala colorimétrica para inspeção visual do grau de hemólise em bolsas de concentrado de hemácias / Colorimetric scale proposal for visual inspection of the hemolysis degree in red cells concentrate bags

Objetivo: Os concentrados de hemácias não estão livres de lesões de estoque, sendo a mais comum a lise das hemácias. Criar uma escala colorimétrica para inspeção visual do grau de hemólise de concentrados de hemácias. Métodos: Foram utilizadas 102 bolsas de concentrados de hemácias (n=36), concentrados de hemácias pobres em leucócitos (n=36) ou de concentrados de hemácias desleucotizadas (n=36), com até 28 dias da produção, em condições padrões de armazenamento e da rotina da Fundação Hemopa. Os valores de hemoglobina e hematócrito foram lidos em contador automatizado. O grau de hemólise foi realizado em amostras de 5 mL do segmento das bolsas, com leitura em espectrofotômetro. O teste de hemólise foi realizado com três gotas de amostras do segmento das bolsas, em tubo seco, adição de 4 mL de solução fisiológica, centrifugação e inspeção visual do sobrenadante (estudo duplo-cego). Para a confecção da escala colorimétrica foram utilizados os resultados de grau de hemólise e teste de hemólise (n=36) e registro fotográfico dos vários níveis de grau de hemólise das bolsas. Para a validação da escala colorimétrica foram realizados grau de hemólise e teste de hemólise para as bolsas restantes (n=66), e com os resultados comparados com a escala proposta (estudo duplo-cego). Resultados: Apenas 7,5% (5/66) das bolsas testadas foram negativas para o teste de hemólise clássico (visual), e positivas quando reavaliadas na escala colorimétrica proposta. Conclusão: A escala colorimétrica proposta mostrou-se simples e reproduzível para definição de teste de hemólise e o grau de hemólise para concentrados de hemácias.

Objective: Red blood cell (RBC) concentrates are not free of stock lesions, the most common of which is lysis of RBC. Create a colorimetric scale for visual inspection of the degree of hemolysis of bag red blood cells. Methods: Eighty-two bags of RBC concentrates (n = 36), RBC concentrates poor in leukocytes (n = 36) or RBC concentrates desleucotizadas (n = 36) were used with up to 28 days of production, under standard storage conditions of the Hemopa Foundation routine. Hemoglobin and hematocrit values were read in automated counter. The degree of hemolysis performed in 5ml samples from the bag segment, with spectrophotometer reading. The hemolysis test was performed with three drops of bag segment samples, in a dry tube, addition of 4 mL of physiological solution, centrifugation and visual inspection of the supernatant (double-blind study). To prepare the colorimetric scale used was the degree of hemolysis and hemolysis test results (n = 36) and photographic records of multiple levels of degree of hemolysis of bags. For the validation of the Colorimetric scale, hemolysis degree and hemolysis test were performed for the remaining bags (n = 66), and the results were compared with the proposed scale (double-blind study). Results: Only 7.5% (5/66) of the tested bags were negative for the classical (visual) hemolysis test, and positive when reevaluated on the proposed colorimetric scale. Conclusion: The proposed colorimetric scale was simple and reproducible for the definition of hemolysis test and the degree of hemolysis for RBC concentrates.