RESUMO
RAD23 (RADIATION SENSITIVE23) proteins are a group of UBL-UBA (ubiquitin-like-ubiquitin-associated) proteins that shuttle ubiquitylated proteins to the 26S proteasome for breakdown. Drought stress is a major environmental constraint that limits plant growth and production, but whether RAD23 proteins are involved in this process is unclear. Here, we demonstrated that a shuttle protein, MdRAD23D1, mediated drought response in apple plants (Malus domestica). MdRAD23D1 levels increased under drought stress, and its suppression resulted in decreased stress tolerance in apple plants. Through in vitro and in vivo assays, we demonstrated that MdRAD23D1 interacted with a proline-rich protein MdPRP6, resulting in the degradation of MdPRP6 by the 26S proteasome. And MdRAD23D1 accelerated the degradation of MdPRP6 under drought stress. Suppression of MdPRP6 resulted in enhanced drought tolerance in apple plants, mainly because the free proline accumulation is changed. And the free proline is also involved in MdRAD23D1-mediated drought response. Taken together, these findings demonstrated that MdRAD23D1 and MdPRP6 oppositely regulated drought response. MdRAD23D1 levels increased under drought, accelerating the degradation of MdPRP6. MdPRP6 negatively regulated drought response, probably by regulating proline accumulation. Thus, "MdRAD23D1-MdPRP6" conferred drought stress tolerance in apple plants.
Assuntos
Malus , Ubiquitina , Ubiquitina/metabolismo , Proteínas de Transporte , Malus/genética , Proteínas de Plantas/genética , Secas , Regulação da Expressão Gênica de Plantas , Estresse Fisiológico , Plantas Geneticamente Modificadas/metabolismoRESUMO
BACKGROUND: TERMINAL FLOWER 1 (TFL1) belongs to the phosphatidylethanolamine-binding protein (PEBP) family, which is involved in inflorescence meristem development and represses flowering in several plant species. In the present study, four TFL1 genes were cloned from the mango (Mangifera indica L.) variety 'SiJiMi' and named MiTFL1-1, MiTFL1-2, MiTFL1-3 and MiTFL1-4. RESULTS: Sequence analysis showed that the encoded MiTFL1 proteins contained a conserved PEBP domain and belonged to the TFL1 group. Expression analysis showed that the MiTFL1 genes were expressed in not only vegetative organs but also reproductive organs and that the expression levels were related to floral development. Overexpression of the four MiTFL1 genes delayed flowering in transgenic Arabidopsis. Additionally, MiTFL1-1 and MiTFL1-3 changed the flower morphology in some transgenic plants. Yeast two-hybrid (Y2H) analysis showed that several stress-related proteins interacted with MiTFL1 proteins. CONCLUSIONS: The four MiTFL1 genes exhibited a similar expression pattern, and overexpression in Arabidopsis resulted in delayed flowering. Additionally, MiTFL1-1 and MiTFL1-3 overexpression affected floral organ development. Furthermore, the MiTFL1 proteins could interact with bHLH and 14-3-3 proteins. These results indicate that the MiTFL1 genes may play an important role in the flowering process in mango.
Assuntos
Arabidopsis/fisiologia , Flores/fisiologia , Mangifera/genética , Proteínas de Plantas/genética , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Flores/genética , Regulação da Expressão Gênica de Plantas , Inflorescência/genética , Proteína de Ligação a Fosfatidiletanolamina/genética , Filogenia , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Técnicas do Sistema de Duplo-HíbridoRESUMO
KEY MESSAGE: BIG regulates the shoot stem cell population. The shoot apical meristem (SAM) contains a population of self-renewing cells, and provides daughter cells for initiation and development of aerial parts of plants. However, the underlying mechanisms of SAM size regulation remain largely unclear. Here, we identified a mutant that displayed a large SAM, designated big-shoot meristem (big-m), in Arabidopsis thaliana. The phenotype of big-m is caused by a new T-DNA insertion allele of BIG, causing a loss of function. The big-m mutant had more stem cells in the SAM than in the wild type. Expression of WUSCHEL (WUS) and SHOOTMERISTEMLESS (STM) was promoted in big-m compared with the wild type, showing that BIG functions upstream of WUS and STM. Therefore, BIG is an important regulator of the stem cell population in the SAM. Furthermore, genetic analysis indicated that BIG acts synergistically with PIN-FORMED1 (PIN1) in controlling SAM size. Our results suggest that BIG plays an important role in controlling Arabidopsis thaliana SAM growth via PIN1-mediated auxin homeostasis.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteínas de Ligação a Calmodulina/metabolismo , Meristema/citologia , Meristema/genética , Brotos de Planta/citologia , Brotos de Planta/genética , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Ligação a Calmodulina/genética , Regulação da Expressão Gênica de Plantas/genética , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismo , Meristema/crescimento & desenvolvimento , Meristema/metabolismo , Mutagênese Insercional , Fenótipo , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Células-Tronco/citologia , Células-Tronco/metabolismoRESUMO
Phosphopeptide enrichment is essential for the phosphoprotein profiling, due to the low abundance in complex biological samples. Moreover, selective binding of multi-phosphopeptides over mono-phosphopeptides is rarely established, but strongly needed in real sample analysis, especially for the investigation of cell behaviors with the multisite phosphorylation cascades. Here two-dimensional (2D) nanosheets were synthesized of Egyptian blue (CaCuSi4 O10 ), the well-known ancient pigment, and its analogues (SrCuSi4 O10 and BaCuSi4 O10 ), which were employed in the enrichment of phosphopeptides for the first time. Surprisingly, the 2D CaCuSi4 O10 nanosheet was highly selective towards multi-phosphopeptides without enriching the mono-phosphorylated peptides in a wide range of acidic conditions or buffer compositions. Meanwhile, the SrCuSi4 O10 and BaCuSi4 O10 nanosheet analogues do not exhibit this unique selectivity. Moreover, the ultrathin and well-defined 2D morphology, with abundant CaII , of Egyptian blue nanosheet was applied in cortical samples of forebrain specific PDK1 conditional knockout mice and their age-matched littermate controls, that are associated with Alzheimer's disease. The as-prepared 2D CaCuSi4 O10 nanosheet not only showed specific selectivity, but also exhibited high sensitivity (detection limit of 4×10-7 m).
Assuntos
Cobre/química , Nanoestruturas/química , Fosfopeptídeos/análise , Silicatos/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Doença de Alzheimer/metabolismo , Doença de Alzheimer/patologia , Animais , Encéfalo/metabolismo , Encéfalo/patologia , Cobre/metabolismo , Nanopartículas Metálicas/química , Camundongos , Camundongos Knockout , Leite/química , Leite/metabolismo , Fosfopeptídeos/química , Fosfopeptídeos/metabolismo , Ligação Proteica , Proteínas Serina-Treonina Quinases/deficiência , Proteínas Serina-Treonina Quinases/genética , Piruvato Desidrogenase Quinase de Transferência de Acetil , Silicatos/metabolismo , Titânio/química , Proteínas tau/química , Proteínas tau/metabolismoRESUMO
In the glucose measuring technique by surface plasmon resonance, D-galactose/D-glucose binding protein (GGBP) that can specifically adsorb glucose was introduced, and high-precision specific detection of glucose concentration was realized. In the present paper, the GGBP protein was bound on the surface of SPR sensor through thiol coupling method. GGBP binding experiment was carried out on SPR sensor and then glucose concentration experiment was conducted with this sensor. The results indicated that the SPR sensor had good linearity, stability and repeatability in the range of 0.1-10 mg x dL(-1). SPR sensor bound with GGBP would have great potential and vast development prospects.
Assuntos
Glucose/análise , Ressonância de Plasmônio de Superfície , Galactose , Proteínas de Transporte de Monossacarídeos , Ligação Proteica , Compostos de SulfidrilaRESUMO
Measuring the glucose concentrations in the interstitial fluid is currently the main method to achieve the continuous blood glucose monitoring. The MIR-ATR(Mid-infrared, Attenuated Total Reflection)Spectroscopy has prominent advantage on the analysis of small biological molecule for composition information like the glucose, but it is still an unresolved problem that how to detect the subcutaneous glucose concentration by using the MIR-ATR Spectroscopy. In the present paper, we carry out the experiment based on MIR-ATR for the detection of subcutaneous glucose information on both the natural state and the penetration state based on the theoryanalysis of MIR penetration depth. Firstly, collect spectral data of the subcutaneous glucose concentration of human finger on the natural state were collected as the light shined the skin directly, and it was discussed whether the MIR can penetrate the skin to get the information of subcutaneous glucose. On this basis, collect spectral data of the subcutaneous glucose concentration of human finger at the penetration state were collected when the Interstitial fluid is permeated to the surface layer by using low-frequency ultrasound and vacuum, then it analyzed that whether it can detect the glucose-specific information or not. As the two-dimensional correlation spectroscopy has high resolution and good versatility, it is widely used to analyze the inter-molecular reaction and judge the absorption peaks information in many fields including the MIR spectroscopy field, so we choose the Two-dimensional correlation spectroscopy to analyze the information of subcutaneous glucose concentration at the natural state and the penetration state. The experiment result shows that the MIR-ATR spectroscopy can't be applied in the detection of subcutaneous glucose concentrationdirectly, and it is a promising direction to make the Interstitial fluid permeated to the surface layer by the physical methods or chemical methods.
Assuntos
Líquido Extracelular/química , Glucose/análise , Humanos , Pele , Espectrofotometria InfravermelhoRESUMO
Comprehensive and deep research on the variations of ecosystem service value (ESV) based on land utilization/land cover change from a spatio-temporal perspective is of great significance for regional ecosystem optimization, as well as coordinated sustainable development of natural environment and economic society. Based on land utilization, natural environment, and socio-economic data of Dongjiang River Basin from 2010 to 2020, combined with hotspot analysis tools and local spatial autocorrelation analysis methods, we comprehensively analyzed the spatio-temporal variations of Dongjiang River Basin ESV, and further explored the spatial differentiation mechanism with geographic detector tools. The results showed that Dongjiang River Basin was dominated by forest ecosystem from 2010 to 2020. The construction land area had expanded significantly mainly from arable land and forest. The Dongjiang River Basin ESV showed a downward and then an upward trend. The ESV of arable land, forest and construction land continuously decreased, and the ESV of water decreased first and then increased substantially. The spatial distribution of ESV hot and cold spots had a significant agglomeration effect, presenting a pattern of hot spot dispersion in the upstream area and cold spots aggregation in the downstream area. The ESV distribution in the upstream and downstream area was not balanced, with the downstream area bearing greater ecological stress. According to the detection results of ESV spatial differentiation mechanism, land utilization was the main factor affecting the spatial differentiation, with the spatial difference of ESV (q value) reaching 0.462. The interaction of factors could greatly strengthen the spatial differentiation effect on Dongjiang River Basin ESV.
Assuntos
Conservação dos Recursos Naturais , Ecossistema , Florestas , China , Análise EspacialRESUMO
The CO/COL gene family plays an important role in regulating photoperiod-dependent flowering time in plants. In this study, two COL2 gene homologs, MiCOL2A and MiCOL2B, were isolated from 'SiJiMi' mango, and their expression patterns and functions were characterized. The MiCOL2A and MiCOL2B genes both belonged to the group â of CO/COL gene family. MiCOL2A and MiCOL2B exhibited distinct circadian rhythms and were highly expressed in leaves during the flowering induction period. Subcellular localization analysis revealed that MiCOL2A and MiCOL2B are localized in the nucleus. The overexpression of MiCOL2A and MiCOL2B significantly delayed flowering time in Arabidopsis under both long-day (LD) and short-day (SD) conditions. The MiCOL2A and MiCOL2B overexpression Arabidopsis plants exhibited more tolerance to slat and drought stress after abiotic stress treatments, with greater ROS scavenging capacity and protective enzyme activity, less cell damage and death and higher expression of stress response genes than wild type plants. Bimolecular fluorescence complementation (BiFC) analysis showed that MiCOL2A and MiCOL2B interacted with several stress-related proteins, including zinc finger protein 4 (MiZFP4), MYB30-INTERACTING E3 LIGASE 1 (MiMIEL1) and RING zinc finger protein 34 (MiRZFP34). The results indicate that MiCOL2A and MiCOL2B are not only involved in flowering time but also play a positive role in abiotic stress responses in plants.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Proteínas de Ligação a DNA , Regulação da Expressão Gênica de Plantas , Mangifera , Plantas Geneticamente Modificadas , Estresse Fisiológico , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Ligação a DNA/genética , Flores/genética , Flores/crescimento & desenvolvimento , Mangifera/genética , Fotoperíodo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Estresse Fisiológico/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
APETALA1 (AP1), CAULIFLOWER (CAL) and FRUITFULL (FUL) were homologous genes with redundant functions in the process of flower transformation and floral development in Arabidopsis. Two CALs genes, MiCAL1 and MiCAL2, were cloned from mango (Mangifera indica L.). Their full-length sequences contained 717 bp and 714 bp, encoding 239 and 238 amino acids, respectively. Both the MiCAL1 and MiCAL2 proteins contained typical MADS-box and K-box domains and therefore belonged to the CAL-like protein family. MiCAL1 and MiCAL2 were expressed in all tissues at the inflorescence elongation stage and flowering stage, with the highest expression in the leaves at the flowering stage. They had similar expression patterns during flower development, with the highest expression levels in leaves during flower differentiation and the lowest expression levels during fruit development. Overexpression of MiCAL1 and MiCAL2 resulted in significantly earlier flowering in Arabidopsis. Overexpression of MiCAL1 resulted in terminal flowers with normal flower organs, while overexpression of MiCAL2 induced partially variation in floral organs but had no effect on inflorescences. Yeast two-hybrid (Y2H) experiments showed that MiCAL1 and MiCAL2 can interact with several flower-related proteins as well as stress response proteins, such as SEP1, SVP1, SVP2, SOC1G and Di19-4. These results suggest that these two MiCALs genes may have an important influence on mango flowering.
Assuntos
Arabidopsis , Brassica , Mangifera , Arabidopsis/metabolismo , Mangifera/genética , Mangifera/metabolismo , Regulação da Expressão Gênica de Plantas , Expressão Ectópica do Gene , Brassica/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Flores , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Proteínas de Domínio MADS/genéticaRESUMO
The CONSTANS-LIKE1 (COL1) gene plays an important role in the regulation of photoperiodic flowering in plants. In this study, two COL1 homolog genes, MiCOL1A and MiCOL1B, were isolated from mango (Mangifera indica L.). The open reading frames of MiCOL1A and MiCOL1B are 852 and 822 bp in length and encode 284 and 274 amino acids, respectively. The MiCOL1A and MiCOL1B proteins contain only one CCT domain and belong to the CO/COL group IV protein family. MiCOL1A and MiCOL1B were expressed both in vegetative and reproductive organs but with expression level differences. MiCOL1A was highly expressed in juvenile and adult leaves, but MiCOL1B was highly expressed in flowers. Seasonal expression analysis showed that MiCOL1A and MiCOL1B have similar expression patterns and higher expression levels during flower induction and flower organ differentiation periods. However, MiCOL1A and MiCOL1B exhibited unstable patterns in circadian expression analysis. MiCOL1A and MiCOL1B were localized in the nucleus and had transcriptional activation activity in yeast. Overexpression of MiCOL1A and MiCOL1B resulted in significantly delayed flowering time in Arabidopsis. Furthermore, we also found that overexpression of MiCOL1A and MiCOL1B enhanced drought tolerance in transgenic Arabidopsis. The results demonstrated that MiCOL1A and MiCOL1B are not only involved in flowering regulation but also play a role in the stress response of plants.
Assuntos
Flores/fisiologia , Mangifera , Proteínas de Plantas , Arabidopsis/genética , Arabidopsis/fisiologia , Regulação da Expressão Gênica de Plantas , Mangifera/genética , Mangifera/fisiologia , Fotoperíodo , Proteínas de Plantas/genética , Proteínas de Plantas/fisiologia , Plantas Geneticamente Modificadas/fisiologiaRESUMO
BACKGROUND: Uncontrolled inflammation causes health problems. Extracellular signal-regulated kinase (ERK) phosphorylates signal transducer and activator of transcription 3 (STAT3) at Ser727, resulting in inflammation. The leaf of Vernonia amygdalina (VA) is a medicinal herb for managing inflammation-associated diseases. Oral administration or topical application of VA leaf extract exerts anti-inflammatory effects in rat models. However, the anti-inflammatory mechanisms of the herb are not fully understood. PURPOSE: In this study, we aimed to investigate the involvement of ERK/STAT3 (Ser727) signaling in the anti-inflammatory effects of an ethanolic extract of VA leaves. STUDY DESIGN AND METHODS: Extracts of VA leaves were prepared with different concentrations of ethanol. A LPS-stimulated RAW264.7 cell model was used for in vitro assays, and a TPA (12-O-tetradecanoylphorbol-13-acetate)-induced ear edema mouse model was employed for in vivo assays. The 95% ethanol extract of VA leaves (VAE) exerted the strongest inhibitory effect on nitric oxide (NO) production in LPS-stimulated macrophages; thus it was selected for use in this study. Hematoxylin and eosin (H&E) staining was used to examine pathological conditions of mouse ear tissues. Griess reagent was employed to examine NO generation in cell cultures. Immunoblotting and ELISA were used to examine protein levels, and RT-qPCR was employed to examine mRNA levels. RESULTS: Topical application of VAE ameliorated mouse ear edema induced by TPA. VAE suppressed the phosphorylation of ERK (Thr202/Tyr204) and STAT3 (Ser727); and decreased protein levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin (IL)-6, IL-1ß and tumor necrosis factor-α (TNF-α) in the mouse ear tissues and in LPS-stimulated RAW 264.7 cells. VAE also inhibited NO production, and lowered mRNA levels of IL-6, IL-1ß and TNF-α in the macrophages. CONCLUSIONS: VAE ameliorates TPA-induced mouse ear edema. Suppression of ERK/STAT3 (Ser727) signaling is involved in VAE's anti-inflammatory effects. These novel data provide further pharmacological justifications for the medicinal use of VA in treating inflammation-associated diseases, and lay the groundwork for developing VAE into a new anti-inflammatory agent.
Assuntos
Fator de Transcrição STAT3 , Vernonia , Animais , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Edema/tratamento farmacológico , Etanol , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Interleucina-6/metabolismo , Lipopolissacarídeos/farmacologia , Camundongos , NF-kappa B/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Extratos Vegetais/uso terapêutico , RNA Mensageiro , Ratos , Fator de Transcrição STAT3/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Histone acetyltransferases are responsible for histone acetylation, while histone deacetylases (HDACs) counteract histone acetylation. An unbalanced dynamic between histone acetylation and deacetylation may lead to aberrant chromatin landscape and chromosomal function. HDAC2, a member of class I HDAC family, serves a crucial role in the modulation of cell signaling, immune response and gene expression. HDAC2 has emerged as a promising therapeutic target for liver disease by regulating gene transcription, chromatin remodeling, signal transduction and nuclear reprogramming, thus receiving attention from researchers and clinicians. The present review introduces biological information of HDAC2 and its physiological and biochemical functions. Secondly, the functional roles of HDAC2 in liver disease are discussed in terms of hepatocyte apoptosis and proliferation, liver regeneration, hepatocellular carcinoma, liver fibrosis and nonalcoholic steatohepatitis. Moreover, abnormal expression of HDAC2 may be involved in the pathogenesis of liver disease, and its expression levels and pharmacological activity may represent potential biomarkers of liver disease. Finally, research on selective HDAC2 inhibitors and noncoding RNAs relevant to HDAC2 expression in liver disease is also reviewed. The aim of the present review was to improve understanding of the multifunctional role and potential regulatory mechanism of HDAC2 in liver disease.
Assuntos
Histona Desacetilase 2/metabolismo , Histonas/metabolismo , Hepatopatias/enzimologia , RNA não Traduzido/genética , Acetilação , Apoptose , Carcinoma Hepatocelular/enzimologia , Carcinoma Hepatocelular/terapia , Proliferação de Células , Hepatócitos/enzimologia , Histona Desacetilase 2/genética , Humanos , Cirrose Hepática/enzimologia , Cirrose Hepática/terapia , Hepatopatias/terapia , Regeneração Hepática , Hepatopatia Gordurosa não Alcoólica/enzimologia , Hepatopatia Gordurosa não Alcoólica/terapiaRESUMO
To examine the suitability of Myzus persicae, Lipaphis erysimi, Semiaphis heraclei and Aphis gossypii to propagation of Harmonia axyridis population, we studied the development and reproduction of this ladybird by constructing two-sex life table in the laboratory with those four aphid species as diet. The results showed that the immature duration of H. axyridis decreased in an order of L. erysimi (18.18 d), A. gossypii (17.48 d), S. heraclei (16.23 d), and M. persicae (15.77 d). The survival rates of preadult period were S. heraclei (88.3%), M. persicae (86.7%), L. ery-simi (55.0%), and A. gossypii (55.0%). The fecundity of those species were S. heraclei (1750.5), M. persicae (1441.5), A. gossypii (1006.3), and L. erysimi (965.2). The longevity of adult ladybird were S. heraclei (78.8 d), M. persicae (63.1 d), A. gossypii (54.3 d), and L. erysimi (48.4 d). The intrinsic rate of increase (rm) of H. axyridis population decreased in an order of M. persicae (0.19), S. heraclei (0.18), L. erysimi (0.14), and A. gossypii (0.14). The net reproduction rates (R0) were S. heraclei (895.83), M. persicae (600.62), L. erysimi (273.47), and A. gossypii (268.33). Among those four aphid species, S. heraclei and M. persicae were more suitable for the propagation of H. axyridis population.
Assuntos
Afídeos , Besouros , Animais , Fertilidade , Tábuas de Vida , ReproduçãoRESUMO
Alcoholic liver injury (ALI) is a part of alcohol-related liver diseases. These diseases include steatohepatitis, alcoholic fibrosis, cirrhosis and hepatocellular carcinoma (HCC). Accumulating data indicates that alcohol metabolism and circulating endotoxin/lipopolysaccharide (LPS) contribute to macrophage activation, which leads to the development of ALI. Protein tyrosine phosphatase 1B (PTP1B) has been shown to be involved in many tissue inflammations as well as liver fibrosis; however, the role of PTP1B in ALI is still unclear. In this study, PTP1B expression was elevated in liver tissues and primary macrophages isolated from EtOH-fed mice. Moreover, PTP1B expression was elevated in RAW264.7 cells stimulated with alcohol and LPS. Additional studies showed that silencing of PTP1B reduced the inflammatory response and expression of inflammatory cytokines such as IL-1ß, IL-6 and TNF-α, while overexpression of PTP1B induced inflammation in RAW264.7 cells. In addition, we found that NF-κB pathway was activated in RAW264.7 cells stimulated with alcohol and LPS, and PTP1B silencing or overexpression could regulate NF-κB signaling. In conclusion, this study revealed the function of PTP1B in ALI via its regulation of the NF-κB signaling pathway and may provide theoretical support for further research on ALI.
Assuntos
Hepatopatias Alcoólicas/genética , Ativação de Macrófagos , NF-kappa B/genética , Proteína Tirosina Fosfatase não Receptora Tipo 1/genética , Transdução de Sinais/genética , Animais , Depressores do Sistema Nervoso Central/farmacologia , Citocinas/biossíntese , Etanol/farmacologia , Lipopolissacarídeos/farmacologia , Fígado/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteína Tirosina Fosfatase não Receptora Tipo 1/biossíntese , Células RAW 264.7 , Regulação para CimaRESUMO
FLOWERING LOCUS T (FT) is a key integrator of environmental signals and internal cues and plays a central role in the photoperiod response mechanism in Arabidopsis. However, the function of FTs in Mangifera indica L. is unknown. In this study, we identified three MiFTs genes from mango and characterized their role in flowering regulation. The open reading frames of MiFT1, MiFT2, and MiFT3 are 540, 516, and 588 bp in length and encode 180, 172, and 196 amino acids, respectively; the genes belong to the PEBP family. MiFTs share the conserved exon/intron structure of FTs. The nucleotide sequence of MiFT1 is 90% identical to that of MiFT2 and 82% identical to that of MiFT3; MiFT2 and MiFT3 share 81% homology with each other. According to expression analysis, MiFTs were detected at different expression levels in all tested tissues. The expression levels of the three MiFTs were significantly different in leaves during flower development, and MiFT1 expression increased sharply in leaves and was significantly higher than that of the other two MiFTs during flower bud development. All three MiFTs showed daily cycles. Ectopic expression of the three MiFTs in transgenic Arabidopsis resulted in an earlier flowering genotype under long-day conditions, and MiFT1 had the strongest effect in promoting flowering. Additionally, overexpression of three MiFTs in Arabidopsis upregulated the expression levels of several flowering-related genes. Our results suggest that the three MiFTs have positive roles in promoting flowering and suggest that MiFT1 may acts as a key regulator in the flowering pathway.
Assuntos
Flores/genética , Genes de Plantas , Mangifera/genética , Proteínas de Plantas/genética , Flores/fisiologia , Regulação da Expressão Gênica de Plantas , Mangifera/fisiologia , Fotoperíodo , Plantas Geneticamente Modificadas/fisiologiaRESUMO
Yes-associated protein (YAP) is a significant downstream protein in the Hippo signaling pathway with important functions in cell proliferation, apoptosis, invasion and migration. YAP also plays a role in the progression and development of various liver diseases. In hepatic fibrosis development and reversion, the proliferation and apoptosis of activated hepatic stellate cells (HSCs) play a critical role. However, the contribution of YAP to hepatic fibrosis progression and reversion and the underlying mechanism have not been investigated. Here we investigated the expression and function of YAP in the proliferation and apoptosis of activated HSCs. We found that YAP expression was increased in liver fibrosis tissues from CCl4-induced model mice and restored to normal level after stopping CCl4 injection and 6 weeks of spontaneously recovery. YAP expression was elevated in HSC-T6 cells treated with TGF-ß1 and recovered after MDI treatment. Silencing of YAP inhibited the activation and proliferation of HSC-T6 cells stimulated by TGF-ß1. In addition, the apoptosis of activated HSC-T6 cells silenced for YAP was slightly enhanced. Furthermore, over-expression of YAP repressed the reversion of activated HSC-T6 cells mediated by MDI reversal. We found that HSC-T6 cells activated by TGF-ß1 showed higher levels of nuclear YAP compared with MDI-treated cells, indicating that YAP was activated in HSC-T6 cells treated by TGF-ß1. We also found that loss of YAP attenuated Wnt/ß-catenin pathway activity in activated HSC-T6 cells. Treatment of VP, an inhibitor of the YAP-TEAD complex, reduced both activation and proliferation of HSC-T6 cells and increased apoptosis. Together these results indicated that reduced expression of YAP contributes to acquisition of the quiescent phenotype in HSCs. Our results suggest that YAP may be a useful target in HSCs activation and reversion.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Apoptose , Células Estreladas do Fígado/metabolismo , Células Estreladas do Fígado/patologia , Cirrose Hepática/metabolismo , Cirrose Hepática/patologia , Fosfoproteínas/metabolismo , Animais , Apoptose/efeitos dos fármacos , Tetracloreto de Carbono , Proteínas de Ciclo Celular , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Modelos Animais de Doenças , Inativação Gênica/efeitos dos fármacos , Células Estreladas do Fígado/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Masculino , Camundongos Endogâmicos C57BL , Modelos Biológicos , Fator de Crescimento Transformador beta1/metabolismo , Verteporfina/farmacologia , Via de Sinalização Wnt/efeitos dos fármacos , Proteínas de Sinalização YAPRESUMO
Photodegradation is one of the key factors that affect bamboo material application in the exterior environment. Photo radiation will cause chemical degradation, discoloration and cracks on the bamboo surface, thus resulting in weakened strength. The study imitated the accelerated weathering effect of Moso bamboo in sunlight by using UV 313 light. Results showed that after UV irradiation, lignin content decreased sharply. Lignin degradation products are commonly rich in double bonds conjugated with benzene rings; they absorb UV light and shift surface spectral absorbency from the visible to the UV region and play an important role in the first stage of reddish-yellow discoloration. The photochemical reactions were very rapid at the beginning and then slowed down after one week. The degraded products covered the surface and protected the inner layer from further degradation. The surface colour turned grey and lighter with erosion of degradation products when the experimental time was extended.
RESUMO
SUMOylation and deSUMOylation, a dynamic process, is proved to be involved in various fibrotic diseases. Here, we found SENP2, one of deSUMOylation protease family member, was decreased in CCl4-induced mice fibrotic liver tissues, primary HSCs and restored after spontaneously recovery. In addition, HSC-T6 cells with TGF-ß1 treatment resulted in a significant reduction of SENP2. Ectopic expression of SENP2 hindered cells activation and proliferation induced by TGF-ß1 while knockdown of SENP2 showed an opposite effect. Importantly, SENP2 promoted apoptosis of HSC-T6 cells activated by TGF-ß1. Furthermore, restoration of SENP2 was observed in inactivated HSCs after adipogenic differentiation mixture (MDI) treatment. Inadequate SENP2 inhibited the reversion of HSC-T6 cells, featured as aberrant expressions of α-SMA and col1a1, two markers of liver fibrosis. It has been reported SENP2 was a suppressant regulator of Wnt/ß-catenin signal pathway. Similarly, we found SENP2 has a negative effect on ß-catenin as well as its downstream genes C-myc and CyclinD1 in liver fibrosis. Collectively, our data indicated SENP2 may be involved in HSCs apoptosis and reversion in liver fibrosis.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Apoptose/efeitos dos fármacos , Cisteína Endopeptidases/metabolismo , Modelos Animais de Doenças , Células Estreladas do Fígado/efeitos dos fármacos , Cirrose Hepática/prevenção & controle , Proteínas Adaptadoras de Transdução de Sinal/antagonistas & inibidores , Proteínas Adaptadoras de Transdução de Sinal/genética , Animais , Biomarcadores/metabolismo , Tetracloreto de Carbono/toxicidade , Linhagem Celular , Proliferação de Células , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Cisteína Endopeptidases/química , Cisteína Endopeptidases/genética , Células Estreladas do Fígado/metabolismo , Células Estreladas do Fígado/patologia , Cirrose Hepática/metabolismo , Cirrose Hepática/patologia , Masculino , Camundongos Endogâmicos C57BL , Interferência de RNA , Distribuição Aleatória , Ratos , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Sumoilação/efeitos dos fármacosRESUMO
The activation of hepatic stellate cells (HSCs) is a central event in the progression of liver fibrosis. Multiple studies proved that DNA methylation might accelerate HSCs activation. However, the specific pathogenesis of liver fibrosis remains not fully addressed. Our laboratory performed Genome methylation screening to find out the methylated gene in mice with liver fibrosis. The pilot experiments showed that the promoter of prostacyclin synthase (PTGIS) gene was hypermethylated in CCl4-induced liver fibrosis mouse model. Moreover, the down-regulated PTGIS expression can be restored by DNMTs-RNAi and 5-aza-2-deoxycytidine (5-azadC), an inhibitor of DNA methyltransferase (DNMTs). Methylation-specific PCR (MSP) showed that the methylation status of PTGIS in HSC-T6 cells cultures with TGF-ß1 (10 ng/mL) was elevated compared with control group. Chromatin immunoprecipitation (ChIP) assay indicated that PTGIS methylation was mainly induced by DNMT1 and DNMT3b. We further investigated the function of PTGIS in liver fibrosis by Recombinant Hepatic-adeno-associated virus (rAAV8)-PTGIS overexpression. The data indicated that overexpression of PTGIS in mouse liver accompanied by elevated apoptosis-related proteins expression in primary HSCs. Conversely, PTGIS silencing mediated by RNAi enhanced the expression of α-SMA and COL1a1 in vitro. Those results illustrated that adding PTGIS expression inhibits the activation of HSCs and alleviates liver fibrosis. Therefore, our study unveils the role of PTGIS in HSCs activation, which may provide a possible explanation for CCl4-mediated liver fibrosis.
RESUMO
The aim of this study was to develop a method combining an online concentration and high-efficiency capillary electrophoresis separation to analyze and detect three compounds (rutin, hyperoside, and chlorogenic acid) in Flos Farfarae. In order to get good resolution and enrichment, several parameters such as the choice of running buffer, pH and concentration of the running buffer, organic modifier, temperature, and separation voltage were all investigated. The optimized conditions were obtained as follows: the buffer of 40 mM NaH2P04-40 mM Borax-30% v/v methanol (pH 9.0); the sample hydrodynamic injection of up to 4 s at 0.5 psi; 20 kV applied voltage. The diode-array detector was used, and the detection wavelength was 364 nm. Based on peak area, higher levels of selective and sensitive improvements in analysis were observed and about 14-, 26-, and 5-fold enrichment of rutin, hyperoside, and chlorogenic acid were achieved, respectively. This method was successfully applied to determine the three compounds in Flos Farfarae. The linear curve of peak response versus concentration was from 20 to 400 µg/ml, 16.5 to 330 µg/mL, and 25 to 500 µg/mL, respectively. The regression coefficients were 0.9998, 0.9999, and 0.9991, respectively.