RESUMEN
Agathisflavone, purified from Cenostigma pyramidale (Tul.) has been shown to be neuroprotective in in vitro models of glutamate-induced excitotoxicity and inflammatory damage. However, the potential role of microglial regulation by agathisflavone in these neuroprotective effects is unclear. Here we investigated the effects of agathisflavone in microglia submitted to inflammatory stimulus in view of elucidating mechanisms of neuroprotection. Microglia isolated from cortices of newborn Wistar rats were exposed to Escherichia coli lipopolysaccharide (LPS, 1 µg/mL) and treated or not with agathisflavone (1 µM). Neuronal PC12 cells were exposed to a conditioned medium from microglia (MCM) treated or not with agathisflavone. We observed that LPS induced microglia to assume an activated inflammatory state (increased CD68, more rounded/amoeboid phenotype). However, most microglia exposed to LPS and agathisflavone, presented an anti-inflammatory profile (increased CD206 and branched-phenotype), associated with the reduction in NO, GSH mRNA for NRLP3 inflammasome, IL1-ß, IL-6, IL-18, TNF, CCL5, and CCL2. Molecular docking also showed that agathisflavone bound at the NLRP3 NACTH inhibitory domain. Moreover, in PC12 cell cultures exposed to the MCM previously treated with the flavonoid most cells preserved neurites and increased expression of ß-tubulin III. Thus, these data reinforce the anti-inflammatory activity and the neuroprotective effect of agathisflavone, effects associated with the control of NLRP3 inflammasome, standing out it as a promising molecule for the treatment or prevention of neurodegenerative diseases.
RESUMEN
Corn silk has been widely used as a nutritional and medicinal supplement due to its pharmacological properties, but there is a lack of studies that correlate the extracts' chemical composition with their biological activities. Herein, we performed the large-scale chemical characterization of corn silk extracts and used chemometrics to correlate the chemical composition with the biological activities of the extracts. Twenty-two metabolites were identified by High-Performance Liquid Chromatography coupled to Mass Spectrometry (HPLC-MS), whereas twelve were identified by Gas Chromatography coupled to Mass Spectrometry (GC-MS). Chemometrics allowed us to discriminate extracts obtained in different organic solvents from in natura and commercial product samples and to pinpoint potential candidate metabolites for the antioxidant and anti-glioma activities. Two flavone glycosides (7 and 8), along with a O-methylated anthocyanidin (26) seems to be the main contributors for the biological activities of the corn silk extracts.
Asunto(s)
Extractos Vegetales , Zea mays , Cromatografía Líquida de Alta Presión , Cromatografía de Gases y Espectrometría de Masas , Metabolómica , SedaRESUMEN
Myelin loss is the hallmark of the demyelinating disease multiple sclerosis (MS) and plays a significant role in multiple neurodegenerative diseases. A common factor in all neuropathologies is the central role of microglia, the intrinsic immune cells of the central nervous system (CNS). Microglia are activated in pathology and can have both pro- and anti-inflammatory functions. Here, we examined the effects of the flavonoid agathisflavone on microglia and remyelination in the cerebellar slice model following lysolecithin induced demyelination. Notably, agathisflavone enhances remyelination and alters microglial activation state, as determined by their morphology and cytokine profile. Furthermore, these effects of agathisflavone on remyelination and microglial activation were inhibited by blockade of estrogen receptor α. Thus, our results identify agathisflavone as a novel compound that may act via ER to regulate microglial activation and enhance remyelination and repair.
Asunto(s)
Antiinflamatorios/farmacología , Biflavonoides/farmacología , Cerebelo/efectos de los fármacos , Microglía/efectos de los fármacos , Vaina de Mielina/metabolismo , Neuroinmunomodulación/efectos de los fármacos , Oligodendroglía/efectos de los fármacos , Remielinización/efectos de los fármacos , Animales , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Cerebelo/inmunología , Cerebelo/metabolismo , Cerebelo/patología , Citocinas/metabolismo , Receptor alfa de Estrógeno/efectos de los fármacos , Receptor alfa de Estrógeno/metabolismo , Femenino , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Lisofosfatidilcolinas/toxicidad , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Microglía/inmunología , Microglía/metabolismo , Microglía/patología , Oligodendroglía/inmunología , Oligodendroglía/metabolismo , Oligodendroglía/patología , Fenotipo , Técnicas de Cultivo de TejidosRESUMEN
Astrocytic tumour cells derived from human (GL-15) and rat (C6) gliomas, as well as non-tumoural astrocytic cells, were exposed to the saponin-rich fraction (SF) from Agave sisalana waste and the cytotoxic effects were evaluated. Cytotoxicity assays revealed a reduction of cell viability that was more intensive in glioma than in non-tumoural cells. The SF induced morphological changes in C6 cells. They were characterised by cytoplasmic vacuole formation associated with increase in the formation of acidic lysosomes. The SF was subjected to purification on Sephadex LH-20, which characterised three probable steroidal saponins (sisalins) by electrospray ionisation mass spectrometry multistage (ESI-MSn). Sisalins from sisal may be responsible for the cytotoxicity, which involves cytoplasmatic vacuole formation and selective action for glioma cells.