RESUMEN
Chitosan and its derivatives such as low molecular weight chitosans (LMWCs) have been reported to exert many biological activities, such as antioxidant and antitumor effects. However, complex and molecular weight dependent effects of chitosan remain controversial and the mechanisms that mediate these complex effects are still poorly defined. This study was carried out to investigate the immunostimulative effect of different molecular weight chitosan in RAW264.7 macrophages. Our data suggested that two LMWCs (molecular weight of 3 kDa and 50 kDa) both possessed immunostimulative activity, which was dependent on dose and, at the higher doses, also on the molecular weight. LMWCs could significantly enhance the the pinocytic activity, and induce the production of tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), interferon-γ (IFN-γ), nitric oxide (NO) and inducible nitric oxide synthase (iNOS) in a molecular weight and concentration-dependent manner. LMWCs were further showed to promote the expression of the genes including iNOS, TNF-α. Taken together, our findings suggested that LMWCs elicited significantly immunomodulatory response through up-regulating mRNA expression of proinflammatory cytokines and activated RAW264.7 macrophage in a molecular weight- and concentration-dependent manner.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Quitosano/farmacología , Citocinas/metabolismo , Macrófagos/efectos de los fármacos , Adyuvantes Inmunológicos/química , Animales , Línea Celular , Quitosano/química , Relación Dosis-Respuesta a Droga , Mediadores de Inflamación/metabolismo , Macrófagos/inmunología , Ratones , Peso Molecular , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , ARN Mensajero/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Regulación hacia Arriba/efectos de los fármacosRESUMEN
In this study, the complete mitochondrial genome of Trachypithecus cristatus was determined using PCR method. The mitochondrial genome is 16,557 bp in length, it containing 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes and a control region. The structural organization and gene order of T. cristatus mtDNA were similar to that of most other vertebrates. The results could provide useful information for further studies on phylogenetics of T. cristatus.
Asunto(s)
Colobinae/genética , Genes Mitocondriales/fisiología , Genoma Mitocondrial/fisiología , Animales , Secuencia de Bases , ADN Mitocondrial/genética , Proteínas Mitocondriales/genética , Datos de Secuencia Molecular , ARN/genética , ARN Mitocondrial , ARN Ribosómico/genética , ARN de Transferencia/genéticaRESUMEN
In the present study, the complete mitochondrial genome of Trachypithecus francoisi was determined using PCR reactions. The mitochondrial genome is 16,544 bp in length, consisting of 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes and a control region. The structural organization and gene order of T. francoisi were equivalent to that of most other vertebrates. The T. francoisi mtDNA could provide useful data for further studies on phylogenetics and conservation genetics of this species.
Asunto(s)
Colobinae/genética , ADN Mitocondrial/genética , Genoma Mitocondrial , Animales , Secuencia de Bases , Filogenia , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADN/veterinariaRESUMEN
Sulfated polysaccharides extracted from brown marine algae have been shown to possess a variety of biological activities. We assessed the potential activity of the sulfated polysaccharide from Sargassum horneri (SP) and its isolated two major components (fraction-1 (F1) and fraction-2 (F2)), on anti-inflammatory activity in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. In the present study, analysis of polysaccharide chemical composition found that the constituent ratios of sulfate ester and fucose in SP and F1 were 4.95% vs 7.6%, and 4.48% vs 55.9%, respectively, suggesting that F1 may be a major sulfated polysaccharide containing fucose. Meanwhile, our findings demonstrated that TNF-α secretion levels were significantly (P<0.05) decreased by SP and F1 treatments in LPS-stimulated RAW264.7 cells in a dose-dependent manner under the preventive and repair experimental models. Pro-/anti-inflammatory (TNF-α/IL-10) cytokines secretion ratios by LPS-stimulated RAW264.7 macrophages were significantly (P<0.05) inhibited by SP and F1 treatments, particularly by F1 (at high dose, 200µg/ml). Moreover, NO release and iNOS activity were significantly (P<0.05) inhibited by F1. Collectively, the present study suggested that purified component, F1 from SP, had strong anti-inflammatory effects on LPS-stimulated RAW264.7 macrophages in the preventive and repair manner through inhibiting TNF-α secretion levels and NO release.
Asunto(s)
Antiinflamatorios/farmacología , Inflamación/tratamiento farmacológico , Polisacáridos/farmacología , Sargassum/química , Animales , Antiinflamatorios/química , Antiinflamatorios/aislamiento & purificación , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Inflamación/inducido químicamente , Interleucina-10/metabolismo , Lipopolisacáridos/toxicidad , Macrófagos/efectos de los fármacos , Ratones , Óxido Nítrico/metabolismo , Polisacáridos/química , Polisacáridos/aislamiento & purificación , Células RAW 264.7/efectos de los fármacos , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
The effects of elevated adiponectin (ADN) plasma levels on food intake, body weight, and lipid deposition of wild-type mice through ADN gene transfer using hydrodynamic based-gene delivery (HD) were investigated. The administration of pTarget/ADN significantly increased the blood ADN levels on days 1, 3, and 7 as well as food intake and body weight. Reverse transcription polymerase chain reaction (RT-PCR) was used to investigate the key-function genes involved in lipid deposition on epididymal fat, gastrocnemius, and extensor digitorum longus on days 1 and 7. The amounts of adipose triglyceride lipase, hormone-sensitive lipase, and lipoprotein lipase mRNA in the three samples significantly increased. We determined sirtuin 1 (SIRT1), forkhead box O3 (FOXO3a), and peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) gene expression and protein level in these samples. The amounts of SIRT1, FOXO3a, and PGC-1α mRNA in epididymal fat, gastrocnemius, and extensor digitorum longus remarkably increased. However, a significant increase in SIRT1 and PGC-1α protein levels was only observed in extensor digitorum longus. These results suggest that high doses of ADN can increase food intake and body weight. Elevated ADN levels may also affect fat deposition on the adipose tissue and skeletal muscles of wild-type mice via SIRT1, FOXO3a, and its downstream targets, including PGC-1α.
Asunto(s)
Adiponectina/biosíntesis , Adiponectina/genética , ADN/genética , Plásmidos/genética , Quinasas de la Proteína-Quinasa Activada por el AMP , Adiponectina/sangre , Tejido Adiposo/metabolismo , Tejido Adiposo Blanco/metabolismo , Animales , Secuencia de Bases , Peso Corporal/genética , Ingestión de Alimentos/genética , Proteína Forkhead Box O3 , Factores de Transcripción Forkhead/genética , Factores de Transcripción Forkhead/metabolismo , Técnicas de Transferencia de Gen , Lipasa/genética , Lipasa/metabolismo , Lípidos/genética , Lipoproteína Lipasa/genética , Lipoproteína Lipasa/metabolismo , Hígado/metabolismo , Masculino , Ratones , Ratones Endogámicos ICR , Datos de Secuencia Molecular , Músculo Esquelético/metabolismo , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma , Proteínas Quinasas/genética , Proteínas Quinasas/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores de Adiponectina/genética , Receptores de Adiponectina/metabolismo , Sirtuina 1/genética , Sirtuina 1/metabolismo , Esterol Esterasa/genética , Esterol Esterasa/metabolismo , Transactivadores/genética , Transactivadores/metabolismo , Factores de TranscripciónRESUMEN
Leptin gene transfer in the liver by hydrodynamic-based gene delivery instead of peptide administration was used to investigate the effects of leptin on muscle mass accretion and lipid accumulation in muscles of wild-type mice. Food intake (P<0.01), body weight (P<0.01) and white adipose tissue (P<0.01) were significantly reduced in the leptin gene-treated group compared with the control group. Moreover, plasma leptin concentration was significantly increased after administration of the mouse leptin gene at a dose of 15 µg per mouse for 1 day (P<0.01) or 1 week (P<0.05). Furthermore, the mRNA abundance of myosin heavy chain type I (MyHC-I), myosin heavy chain type II (MyHC-IIa, MyHC-IIx), adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL) and lipoprotein lipase (LPL) genes in gastrocnemius muscle and extensor digitorum longus after administration of leptin for 1 week were significantly increased compared with the control group. Finally, we investigated the mechanism by which leptin gene transfer affects fibromuscular and fat deposition in muscle. Gene expression and protein levels of SIRT1, and proliferator-activated receptor-γ coactivator-1α (PGC-1α) were remarkably increased in extensor digitorum longus. On the other hand, PGC-1α and FOXO3a gene expression was observed to have significantly increased in gastrocnemius muscle. However, only changes in the protein levels of PGC-1α were observed (P<0.05). These results suggest that leptin may affect the growth and development of muscle, and fat deposition in wild-type mice via SIRT1 and FOXO3a and their downstream targets, including PGC-1α.