Composición lipídica y evaluación de las actividades antioxidante y leishmanicida del basidiomiceto Ganoderma sp / Lipid composition and evaluation of the antioxidant and leishmanicidal activities of the basidiomycete Ganoderma sp

Introducción: los hongos del género Ganoderma han sido utilizados para el cuidado de la salud en la medicina tradicional asiática por más de 2000 años. Desde 1980 los estudios químicos han reportado un sin número de metabolitos secundarios con propiedades bioactivas. Objetivo: identificar compuestos lipídicos en el extracto etanólico del hongo Ganoderma sp., además de evaluar sus actividades antioxidante y leishmanicida. Métodos: la extracción de las fracciones lipídicas presentes en el cuerpo fructífero de Ganoderma sp. Se realizó por Cromatografía en Columna. La elucidación estructural se determinó por Espectrometría de Masas y Resonancia Magnética Nuclear. La actividad antioxidante del extracto etanólico fue evaluada con las metodologías del radical 2,2-difenil-1-picrilhidrazil (DPPH) y el radical catiónico 2,2'-azinobis (3-etilbenzotiazolina-6-ácido sulfónico) (ABTS); la actividad leishmanicida por citometría de flujo y la actividad citotóxica usando el ensayo colorimétrico de bromuro de 3-(4,5-dimetil-tiazol-2-il)-2,5-difenil tetrazolio (MTT) sobre la línea celular U937. Resultados: diecinueve esteres metílicos y ergosterol fueron identificados por espectrometría de masas en el extracto etanólico. Un compuesto triterpenoidal se elucidó usando Espectroscopia de Resonancia Magnética Nuclear. Los valores de concentración media inhibitoria (IC 50) de la actividad antioxidante del extracto etanólico para las metodologías de los radicales DPPH y ABTS fueron de 85,63 µg/mL y 62,82 µg/mL, respectivamente. Los valores de las actividades citotóxica y leishmanicida fueron > 200,0 µg/mL y 21,5 µg/mL ± 4,4 respectivamente. Conclusiones: las estructuras de los derivados de ácidos grasos elucidados corresponden a compuestos con diferentes grados de insaturación. En este estudio se realizó el reporte de la Ganoderona A, como compuesto triterpenoidal. La elevada actividad antioxidante en relación a otros trabajos sugiere que este organismo es una fuente importante de metabolitos secundarios con propiedades captadoras de radicales libres, aunque los valores de actividad leishmanicida no fueron significativos se recomienda continuar con el estudio de otras particiones del extracto etanólico(AU)

Introduction: Fungi from the genus Ganoderma have been used in Asian traditional medicine for more than 2 000 years. Since the year 1980 chemical studies have reported a large number of secondary metabolites with bioactive properties. Objective: Identify lipid compounds in ethanolic extract from the fungus Ganoderma sp. and evaluate their antioxidant and leishmanicidal activities. Methods: Extraction of lipid fractions from the fruiting body of Ganoderma sp. was conducted by column chromatography. Structural features were determined by mass spectrometry and nuclear magnetic resonance. Antioxidant activity of the ethanolic extract was evaluated with the methodologies for radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) and cationic radical 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS); leishmanicidal activity by flow cytometry, and cytotoxic activity with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide colorimetric assay (MTT) on cell line U937. Results: Nineteen methyl esters and ergosterol were identified by mass spectrometry in the ethanolic extract. A triterpenoid compound was identified by nuclear magnetic resonance spectroscopy. Mean inhibitory concentration values (IC50) for antioxidant activity of the ethanolic extract using the methodologies for radicals DPPH and ABTS were 85.63 µg/ml and 62.82 µg/ml, respectively. Values for cytotoxic and leishmanicidal activities were > 200.0 µg/ml and 21.5 µg/ml ± 4.4, respectively. Conclusions: The structure of the fatty acid derivatives identified corresponds to compounds with varying degrees of unsaturation. The study included the report of Ganoderma A as a triterpenoid compound. Antioxidant activity was found to be higher than in previous studies, suggesting that this organism is an important source of secondary metabolites with free radical scavenging properties. Although leishmanicidal activity values were not found to be significant, it is recommended to study other partitions of the ethanolic extract(AU)

Repressive effects of RNA interference technique on expression and function of glucocorticoid receptor in human macrophage cell line U937 / 第三军医大学学报

Objective To establish a glucocorticoid receptor knockdown model of human macrophage cell line U937 with RNA interference technique. Methods Two RNAi recombinant plasmids (named pSilencer 3.1-GR 1 and pSilencer 3.1-GR 2) targeting to GR gene were constructed. After RNAi recombinant plasmids were transfected into human macrophage cell line U937, the expressions of GR mRNA and GR protein were evaluated with RT-PCR and Western blotting respectively. The transcriptional activation function of GR was evaluated through the detection of relative luciferase activity after dexamethasone treatment. Results Two RNAi recombinant expression plamids were constructed and identified by sequencing. pSilencer 3.1-GR 2 transfection could inhibit not only GR mRNA and protein expressions, but also transcriptional activation function of GR specially; pSilencer 3.1-GR 1 transfection had no significant changes as compared to normal control. Conclusion A glucocorticoid receptor knockdown model has been established successfully, which offers a new method for the further research of GR biological functions.