RESUMEN
BACKGROUND: Osteoarthritis (OA) treatment aims to improve inflammation and delay cartilage degeneration. However, there is no effective strategy presently available. Ononin, a representative isoflavone glycoside component extracted from natural Chinese herbs, exerts anti-inflammatory and proliferative effects. However, the therapeutic effect of ononin on chondrocyte inflammation remains unclear. METHODS: In this study, we explored the therapeutic effect and potential mechanism of ononin in OA by establishing an interleukin-1 beta (IL-1ß)-induced chondrocyte inflammation model. RESULTS: Our results verified that ononin alleviated the IL-1ß-induced decrease in chondrocyte viability, attenuated the overexpression of the inflammatory factors tumour necrosis factor α (TNF-α) and interleukin 6 (IL-6), and simultaneously inhibited the expression of cartilage extracellular matrix (ECM)-degrading enzymes such as matrix metalloproteinase-13 (MMP-13). Furthermore, the decomposition of Collagen II protein could be alleviated in the OA model by ononin. Finally, ononin improved chondrocyte inflammation by downregulating the mitogen-activated protein kinase (MAPK) and nuclear factor kappa-B (NF-κB) signalling pathways. CONCLUSION: Our findings suggested that ononin could inhibit the IL-1ß-induced proinflammatory response and ECM degradation in chondrocytes by interfering with the abnormal activation of the MAPK and NF-κB pathways, indicating its protective effect against OA.
Asunto(s)
Cartílago/efectos de los fármacos , Glucósidos/farmacología , Inflamación/metabolismo , Interleucina-1beta/metabolismo , Isoflavonas/farmacología , Proteínas Quinasas Activadas por Mitógenos/metabolismo , FN-kappa B/metabolismo , Osteoartritis , Animales , Antiinflamatorios/farmacología , Antiinflamatorios/uso terapéutico , Cartílago/citología , Cartílago/metabolismo , Cartílago/patología , Condrocitos/efectos de los fármacos , Condrocitos/metabolismo , Condrocitos/patología , Regulación hacia Abajo , Glucósidos/uso terapéutico , Inflamación/tratamiento farmacológico , Isoflavonas/uso terapéutico , Sistema de Señalización de MAP Quinasas , Masculino , Metaloproteinasa 13 de la Matriz/metabolismo , Osteoartritis/tratamiento farmacológico , Osteoartritis/metabolismo , Osteoartritis/patología , Fitoterapia , Extractos Vegetales/farmacología , Extractos Vegetales/uso terapéutico , Ratas Sprague-Dawley , Transducción de Señal , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Previous studies have shown that the SUP genes play important roles in flower development and plant growth and morphogenesis. In this study, we isolated and characterized a SUPERMAN-like gene DgSZFP from chrysanthemum. DgSZFP contains one conserved Cys2/His2-type zinc finger motifs in the N-terminal region and an EAR-box in C-terminus. Its expression was significantly higher in nodes, flower buds, disc stamens, and petals than in the other tissues. Overexpression of DgSZFP in tobacco resulted in enhanced branching, reduced plant height, increased the width of petal tubes, produced the staminoid petals and petaloid stamens in flowers, and enhanced the seed weight and size. In addition, DgSZFP-overexpression tobacco plants accumulated high concentrations of cytokinin and chlorophyll. These results suggest that DgSZFP may be the candidate gene for regulating branching and floral organ development in chrysanthemum.
Asunto(s)
Chrysanthemum/genética , Flores/crecimiento & desarrollo , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Nicotiana/genética , Tallos de la Planta/crecimiento & desarrollo , Factores de Transcripción/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Clorofila/genética , Clorofila/metabolismo , Chrysanthemum/crecimiento & desarrollo , Chrysanthemum/metabolismo , Citocininas/genética , Citocininas/metabolismo , Meristema/crecimiento & desarrollo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente , Semillas/crecimiento & desarrollo , Nicotiana/crecimiento & desarrollo , Nicotiana/metabolismo , Factores de Transcripción/metabolismo , Dedos de ZincRESUMEN
The plant-specific NAC (for NAM, ATAF1, 2 and CUC2) transcription factors (TFs) have been implicated in different cellular processes involved in stress responses such as cold, high salinity or drought as well as abscisic acid (ABA) signalling. However, the roles of the chrysanthemum NAC TF genes in plant stress responses are still unclear. A full-length cDNA designated DgNAC1, containing a highly conserved N-terminal DNA-binding NAC domain, has been isolated from chrysanthemum by RACE (rapid amplification of cDNA ends). It encodes a protein of 284 amino acids residues (=~32.9 kDa) and theoretical pI of 7.13. The transcript of DgNAC1 was enriched in roots and flowers than in stems and leaves of the adult chrysanthemum plants. The gene expression was strongly induced by ABA, NaCl, drought and cold treatment in the seedlings. Subcellular localization revealed that DgNAC1:GFP fusion protein was preferentially distributed to nucleus. To assess whether DgNAC1 is a practically useful target gene for improving the stress tolerance of chrysanthemum, we ectopically over-expressed the full-length DgNAC1 cDNA in tobacco and found that the 35S:DgNAC1 transgenic tobacco exhibited a markedly increased tolerance to salt. Despite this increased salt stress tolerance, the transgenic tobacco showed no detectable phenotype defects under normal growth conditions. These results proposed that DgNAC1 is appropriate for application in genetic engineering strategies aimed at improving salt stress tolerance in chrysanthemum.
Asunto(s)
Chrysanthemum/genética , Nicotiana/genética , Nicotiana/metabolismo , Proteínas de Plantas/biosíntesis , Tolerancia a la Sal/fisiología , Factores de Transcripción/biosíntesis , Secuencia de Aminoácidos , Secuencia de Bases , Núcleo Celular/genética , Núcleo Celular/metabolismo , Datos de Secuencia Molecular , Filogenia , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente , Tolerancia a la Sal/genética , Alineación de Secuencia , Transducción de Señal , Cloruro de Sodio , Factores de Transcripción/genéticaRESUMEN
A Cys2/His2-type zinc finger protein gene, DgZFP, was isolated from chrysanthemum by rapid amplification of cDNA ends (RACE) approach. The DgZFP encodes a protein of 211 amino acids residues with a calculated molecular mass of 22.9 kDa and theoretical isoelectric point is 8.59. DgZFP contains two Cys2/His2-type zinc finger motifs, one nuclear localization domain, one Leu-rich domain, and one ethylene-responsive element-binding factor (ERF)-associated amphiphilic repression (EAR) domain. The transcript of DgZFP was enriched in flowers than in roots, stems, and leaves of the adult chrysanthemum plants. The gene expression was strongly induced by NaCl, drought and cold treatment, and weakly by ABA treatment in the seedlings. Subcellular localization revealed that DgZFP was localized preferentially distributed to nucleus. Overexpression of DgZFP improved salt tolerance and resulted in growth suppression in transgenic tobacco. We argued that DgZFP is a new member of the Cys2/His2-type zinc finger protein genes, and it maybe function as a regulator in response to salt stress in plants.