RESUMEN
A novel cyclic dipeptide, 14-hydroxy-cyclopeptine (1), was purified from a deep sea derived fungal isolate identified as an Aspergillus sp. The structure was elucidated by detailed spectroscopic analyses using 1D and 2D NMR experiments and high resolution mass spectrometry. The absolute configuration of the amino acid was determined by Marfey's method. Two conformational isomers of 1 were established by ROE analyses. 1 inhibited nitric oxide production with IC50 values at 40.3 µg/mL in a lipopolysaccharide and recombinant mouse interferon-γ -activated macrophage-like cell line, RAW 264.7 and showed no cytotoxic effect in the tested dose range up to 100 µg/mL.
Asunto(s)
Aspergillus/química , Dipéptidos/química , Dipéptidos/farmacología , Evaluación Preclínica de Medicamentos/métodos , Péptidos Cíclicos/química , Animales , Aspergillus/aislamiento & purificación , Línea Celular/efectos de los fármacos , Dipéptidos/aislamiento & purificación , Concentración 50 Inhibidora , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Espectroscopía de Resonancia Magnética , Ratones , Óxido Nítrico/metabolismo , Péptidos Cíclicos/aislamiento & purificación , Péptidos Cíclicos/farmacología , Conformación Proteica , Agua de Mar/microbiologíaRESUMEN
Here, we investigated the impact of mulberry fruit (MBF) extracts on lipopolysaccharide (LPS)-induced inflammatory responses in RAW 264.7 macrophages, and the therapeutic efficacy of MBF diet in mice with dextran sulfate sodium (DSS)-induced acute colitis and MUC2(-/-) mice with colorectal cancer. In vitro, LPS-induced nitric oxide (NO) production was significantly inhibited by MBF extracts via suppressing the expression of proinflammatory molecules, including inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-1 beta (IL-ß) and IL-6. Particularly, a dose-dependent inhibition on LPS-induced inflammatory responses was observed following treatment with MBF dichloromethane extract (MBF-DE), in which linoleic acid and ethyl linolenate were identified as two active compounds. Moreover, we elucidated that MBF-DE attenuated LPS-induced inflammatory responses by blocking activation of both NF-κB/p65 and pERK/MAPK pathways. In vivo, DSS-induced acute colitis was significantly ameliorated in MBF-fed mice as gauged by weight loss, colon morphology and histological damage. In addition, MBF-fed MUC2(-/-) mice displayed significant decrease in intestinal tumor and inflammation incidence compared to control diet-fed group. Overall, our results demonstrated that MBF suppressed the development of intestinal inflammation and tumorgenesis both in vitro and in vivo, and supports the potential of MBF as a therapeutic functional food for testing in human clinical trials.
Asunto(s)
Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Frutas/química , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Morus/química , FN-kappa B/metabolismo , Extractos Vegetales/farmacología , Transducción de Señal/efectos de los fármacos , Animales , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Transformación Celular Neoplásica/efectos de los fármacos , Transformación Celular Neoplásica/genética , Transformación Celular Neoplásica/metabolismo , Colitis/tratamiento farmacológico , Colitis/etiología , Colitis/metabolismo , Colitis/patología , Neoplasias Colorrectales/etiología , Neoplasias Colorrectales/metabolismo , Citocinas/genética , Citocinas/metabolismo , Sulfato de Dextran/efectos adversos , Suplementos Dietéticos , Modelos Animales de Enfermedad , Regulación de la Expresión Génica/efectos de los fármacos , Mediadores de Inflamación/metabolismo , Ácido Linoleico/química , Ácidos Linolénicos/química , Lipopolisacáridos/efectos adversos , Lipopolisacáridos/inmunología , Ratones , Ratones Noqueados , Mucina 2/deficiencia , Óxido Nítrico/biosíntesis , Fosforilación , Extractos Vegetales/administración & dosificación , Extractos Vegetales/química , Transporte de ProteínasRESUMEN
Edible berries have a broad spectrum of biomedical functions, including improving immune responses and reducing risk for chronic diseases. In this study, the anti-inflammatory activities of crude extracts (CEs), anthocyanin-rich fractions (ARFs), and des-anthocyanin fractions (DAFs) from seven berries were evaluated based on their inhibitory effects on nitric oxide (NO) production in lipopolysaccharide (LPS)/IFN-γ-activated RAW264.7 macrophages. ARFs from red raspberries (RR-ARFs) exhibited the highest efficiency in suppressing NO synthesis. The anti-inflammatory properties were also demonstrated by reducing the expression levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-1 beta (IL-1ß) and IL-6 in RAW264.7 cells. The luciferase reporter assay demonstrated that the activities of NF-κB and AP-1 signaling pathways were significantly suppressed by RR-ARFs. Further studies showed that RR-ARFs decreased the phosphorylation of IKK, IκBα, p65 and JNK and the nuclear translocation of p65 in LPS/IFN-γ-stimulated RAW264.7 cells. In a mouse colitis model, dextran sulfate sodium (DSS)-induced weight loss and histological damage were significantly ameliorated by RR-ARFs treatment. Taken together, our results indicate that RR-ARFs attenuate inflammation both in vitro and in vivo primarily by inhibiting the activation of NF-κB and MAPKs. The anti-inflammatory of RR-ARFs could be harnessed and applied in animal agriculture, drug and food industries.
Asunto(s)
Antocianinas/farmacología , Antiinflamatorios/farmacología , Colitis/inmunología , Extractos Vegetales/farmacología , Rubus/química , Animales , Línea Celular , Supervivencia Celular/efectos de los fármacos , Colitis/tratamiento farmacológico , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/metabolismo , Sulfato de Dextran , Frutas/química , Expresión Génica , Interferón gamma/fisiología , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Lipopolisacáridos/farmacología , MAP Quinasa Quinasa 4/metabolismo , Ratones Endogámicos BALB C , Morus/química , Óxido Nítrico/biosíntesis , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismoRESUMEN
A novel human zinc finger gene, ZNF300, was isolated from human embryos on the base of the enrichment of C2H2-specific mRNAs. The ZNF300 cDNA is 3104 bp long and encodes a 604-amino acid protein with an amino-terminal KRAB domain, and 12 carboxyl-terminal C2H2 zinc finger motifs. According to the genome database, ZNF300 is assigned to chromosome 5q33.1. Northern blot analysis revealed that the ZNF300 transcripts are expressed mostly in heart, skeletal muscle and brain. ZNF300 is localized to the nucleus and the KRAB domain of ZNF300 protein exhibits transcription repressor activity. These results suggest that ZNF300 is a ubiquitous transcription repressor in the nucleus.