RESUMO
BACKGROUND: The hypoxic tumor microenvironment is a key factor that promotes metabolic reprogramming and vascular mimicry (VM) in ovarian cancer (OC) patients. ESM1, a secreted protein, plays an important role in promoting proliferation and angiogenesis in OC. However, the role of ESM1 in metabolic reprogramming and VM in the hypoxic microenvironment in OC patients has not been determined. METHODS: Liquid chromatography coupled with tandem MS was used to analyze CAOV3 and OV90 cells. Interactions between ESM1, PKM2, UBA2, and SUMO1 were detected by GST pull-down, Co-IP, and molecular docking. The effects of the ESM1-PKM2 axis on cell glucose metabolism were analyzed based on an ECAR experiment. The biological effects of the signaling axis on OC cells were detected by tubule formation, transwell assay, RTâPCR, Western blot, immunofluorescence, and in vivo xenograft tumor experiments. RESULTS: Our findings demonstrated that hypoxia induces the upregulation of ESM1 expression through the transcription of HIF-1α. ESM1 serves as a crucial mediator of the interaction between PKM2 and UBA2, facilitating the SUMOylation of PKM2 and the subsequent formation of PKM2 dimers. This process promotes the Warburg effect and facilitates the nuclear translocation of PKM2, ultimately leading to the phosphorylation of STAT3. These molecular events contribute to the promotion of ovarian cancer glycolysis and vasculogenic mimicry. Furthermore, our study revealed that Shikonin effectively inhibits the molecular interaction between ESM1 and PKM2, consequently preventing the formation of PKM2 dimers and thereby inhibiting ovarian cancer glycolysis, fatty acid synthesis and vasculogenic mimicry. CONCLUSION: Our findings demonstrated that hypoxia increases ESM1 expression through the transcriptional regulation of HIF-1α to induce dimerization via PKM2 SUMOylation, which promotes the OC Warburg effect and VM.
Assuntos
Proteínas de Transporte , Ácidos Graxos , Proteínas de Membrana , Proteínas de Neoplasias , Neoplasias Ovarianas , Proteínas de Ligação a Hormônio da Tireoide , Hormônios Tireóideos , Microambiente Tumoral , Feminino , Humanos , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/patologia , Neoplasias Ovarianas/genética , Animais , Hormônios Tireóideos/metabolismo , Camundongos , Proteínas de Membrana/metabolismo , Proteínas de Membrana/genética , Linhagem Celular Tumoral , Ácidos Graxos/metabolismo , Proteínas de Neoplasias/metabolismo , Proteínas de Neoplasias/genética , Proteínas de Transporte/metabolismo , Proteínas de Transporte/genética , Efeito Warburg em Oncologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Regulação Neoplásica da Expressão Gênica , Neovascularização Patológica/metabolismo , Neovascularização Patológica/genética , Neovascularização Patológica/patologia , Ensaios Antitumorais Modelo de Xenoenxerto , Proliferação de Células , ProteoglicanasRESUMO
BACKGROUND: Ovarian cancer (OC) is a malignant neoplasm that displays increased vascularization. Angiopoietin-like 4 (ANGPTL4) is a secreted glycoprotein that functions as a regulator of cell metabolism and angiogenesis and plays a critical role in tumorigenesis. However, the precise role of ANGPTL4 in the OC microenvironment, particularly its involvement in angiogenesis, has not been fully elucidated. METHODS: The expression of ANGPTL4 was confirmed by bioinformatics and IHC in OC. The potential molecular mechanism of ANGPTL4 was measured by RNA-sequence. We used a series of molecular biological experiments to measure the ANGPTL4-JAK2-STAT3 and ANGPTL4-ESM1 axis in OC progression, including MTT, EdU, wound healing, transwell, xenograft model, oil red O staining, chick chorioallantoic membrane assay and zebrafish model. Moreover, the molecular mechanisms were confirmed by Western blot, Co-IP and molecular docking. RESULTS: Our study demonstrates a significant upregulation of ANGPTL4 in OC specimens and its strong association with unfavorable prognosis. RNA-seq analysis affirms that ANGPTL4 facilitates OC development by driving JAK2-STAT3 signaling pathway activation. The interaction between ANGPTL4 and ESM1 promotes ANGPTL4 binding to lipoprotein lipase (LPL), thereby resulting in reprogrammed lipid metabolism and the promotion of OC cell proliferation, migration, and invasion. In the OC microenvironment, ESM1 may interfere with the binding of ANGPTL4 to integrin and vascular-endothelial cadherin (VE-Cad), which leads to stabilization of vascular integrity and ultimately promotes angiogenesis. CONCLUSION: Our findings underscore that ANGPTL4 promotes OC development via JAK signaling and induces angiogenesis in the tumor microenvironment through its interaction with ESM1.
Assuntos
Cistadenocarcinoma Seroso , Janus Quinase 2 , Neoplasias Ovarianas , Fator de Transcrição STAT3 , Animais , Feminino , Humanos , Microambiente Tumoral , Simulação de Acoplamento Molecular , Angiogênese , Peixe-Zebra/metabolismo , Carcinogênese , Proliferação de Células , Carcinoma Epitelial do Ovário , Neoplasias Ovarianas/genética , Linhagem Celular Tumoral , Proteína 4 Semelhante a Angiopoietina/genética , Proteínas de Neoplasias , ProteoglicanasRESUMO
Acidic tea (Camellia sinensis) plantation soil usually suffers from magnesium (Mg) deficiency, and as such, application of fertilizer containing Mg can substantially increase tea quality by enhancing the accumulation of nitrogen (N)-containing chemicals such as amino acids in young tea shoots. However, the molecular mechanisms underlying the promoting effects of Mg on N assimilation in tea plants remain unclear. Here, both hydroponic and field experiments were conducted to analyze N, Mg, metabolite contents, and gene expression patterns in tea plants. We found that N and amino acids accumulated in tea plant roots under Mg deficiency, while metabolism of N was enhanced by Mg supplementation, especially under a low N fertilizer regime. 15N tracing experiments demonstrated that assimilation of N was induced in tea roots following Mg application. Furthermore, weighted gene correlation network analysis (WGCNA) analysis of RNA-seq data suggested that genes encoding glutamine synthetase isozymes (CsGSs), key enzymes regulating N assimilation, were markedly regulated by Mg treatment. Overexpression of CsGS1.1 in Arabidopsis (Arabidopsis thaliana) resulted in a more tolerant phenotype under Mg deficiency and increased N assimilation. These results validate our suggestion that Mg transcriptionally regulates CsGS1.1 during the enhanced assimilation of N in tea plant. Moreover, results of a field experiment demonstrated that high Mg and low N had positive effects on tea quality. This study deepens our understanding of the molecular mechanisms underlying the interactive effects of Mg and N in tea plants while also providing both genetic and agronomic tools for future improvement of tea production.
Assuntos
Camellia sinensis , Camellia sinensis/genética , Camellia sinensis/metabolismo , Magnésio/metabolismo , Glutamato-Amônia Ligase/genética , Glutamato-Amônia Ligase/metabolismo , Nitrogênio/metabolismo , Fertilizantes , Aminoácidos/metabolismo , Chá/metabolismo , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismoRESUMO
Understanding soil organic carbon (SOC), the largest carbon (C) pool of a terrestrial ecosystem, is essential for mitigating climate change. Currently, the spatial patterns and drivers of SOC in the plantations of tea, a perennial leaf crop, remain unclear. Therefore, the present study surveyed SOC across the main tea-producing areas of China, which is the largest tea producer in the world. We analyzed the soil samples from tea plantations under different scenarios, such as provinces, regions [southwest China (SW), south China (SC), south Yangtze (SY), and north Yangtze (NY)], climatic zones (temperate, subtropical, and tropical), and cultivars [large-leaf (LL) and middle or small-leaf (ML) cultivars]. Preliminary analysis revealed that most tea-producing areas (45%) had SOC content ranging from 10 to 20 g kg-1. The highest SOC was recorded for Yunnan among the various provinces, the SW tea-producing area among the four regions, the tropical region among the different climatic zones, and the areas with LL cultivars compared to those with ML cultivars. Further Pearson correlation analysis demonstrated significant associations between SOC and soil variables and random forest modeling (RF) identified that total nitrogen (TN) and available aluminum [Ava(Al)] of soil explained the maximum differences in SOC. Besides, a large indirect effect of geography (latitude and altitude) on SOC was detected through partial least squares path modeling (PLS-PM) analysis. Thus, the study revealed a high spatial heterogeneity in SOC across the major tea-producing areas of China. The findings also serve as a basis for planning fertilization strategies and C sequestration policies for tea plantations.
RESUMO
Tea plants are more sensitive to variations in calcium concentration compared to other plants, whereas a moderate aluminum concentration facilitates the growth and development of tea plants. Aluminum and calcium show a competitive interaction with respect to the uptake of elements, consequently exerting physiological effects on plants. To further explore these interactions, in this study, we used the solution culture method to treat tea plants with two calcium concentrations (0.8 mM and 5.6 mM) and three aluminum concentrations (0 mM, 0.4 mM, and 1 mM). We then determined the influence of the combined treatments on root growth and quality compound accumulation in the tissues by a combination of phenotype, gene expression, and metabolite analyses. Moderate aluminum supplementation (0.4 mM) alleviated the inhibition of root growth caused by high calcium stress. High calcium stress significantly inhibited the accumulation of most amino acids (e.g., Glutamic acid, Citulline, and Arginine) and organic acids (e.g., a-ketoglutaric acid) in the roots, stems, and leaves, whereas aluminum deficiency significantly increased most amino acids in the roots and leaves (except Serine, Alanine, and Phenylalanine in the roots and Ser in the leaves), with a more than two-fold increase in Arg and Lysine. High calcium stress also induced the accumulation of secondary metabolites such as epigallocatechin gallate and procyanidin in the roots, whereas aluminum supplementation significantly reduced the contents of flavonol glycosides such as quercetin, rutin, myricitrin, and kaempferitrin, as well as caffeine, regardless of calcium concentration. Aluminum supplementation reversed some of the changes in the contents of leaf metabolites induced by calcium stress (e.g., 4-dihydroquercetin, apigenin C-pentoside, phenethylamine, and caffeine). Overall, calcium stress caused severe growth inhibition and metabolic disorders in tea plants, which could be reversed by aluminum supplementation, particularly in maintaining the root tips and the accumulation of secondary metabolites. These results provide a theoretical basis for improving calcium-aluminum nutrient management to promote tea plant growth and quality.
Assuntos
Alumínio , Cálcio , Alumínio/toxicidade , Cafeína , Cálcio da Dieta , Aminoácidos , Suplementos Nutricionais , CháRESUMO
To reveal the mechanisms underlying how light affects flavonoid metabolism and the potential role of flavonoids in protecting against photooxidative stress in tea leaves, tea plants adapted to low-light conditions were exposed to full sunlight over 48 h. There was an increase in the activities of catalase (CAT) and superoxide dismutase (SOD) as well as greater accumulation of reactive oxygen species, lutein, tocopherols, ascorbate and malondialdehyde, suggestive of a time-dependent response to photooxidative stress in tea leaves. Analysis of the time dependency of each element of the antioxidant system indicated that carotenoids and tocopherols exhibited the fastest response to light stress (within 3 h), followed by SOD, CAT and catechin, which peaked at 24 h. Meanwhile, flavonols, vitamin C and glutathione showed the slowest response. Subsequent identification of the main phytochemicals involved in protecting against oxidative stress using untargeted metabolomics revealed a fast and initial accumulation of nonesterified catechins that preceded the increase in flavonol glycosides and catechin esters. Gene expression analysis suggested that the light-induced accumulation of flavonoids was highly associated with the gene encoding flavonol synthase. Ultraviolet B (UV-B) irradiation further validated the time-dependent and collaborative effects of flavonoids in photoprotection in tea plants. Intriguingly, the dynamics of the metabolic response are highly distinct from those reported for Arabidopsis, suggesting that the response to light stress is not conserved across plants. This study additionally provides new insights into the functional role of flavonoids in preventing photooxidative stress and may contribute to further improving tea quality through the control of light intensity.
Assuntos
Arabidopsis , Camellia sinensis , Catequina , Flavonoides/metabolismo , Antioxidantes/metabolismo , Catequina/metabolismo , Luz Solar , Folhas de Planta/metabolismo , Camellia sinensis/metabolismo , Arabidopsis/metabolismo , Chá/metabolismo , Tocoferóis/metabolismo , Superóxido Dismutase/metabolismoRESUMO
RING finger protein 6 (RNF6), a RING finger protein, has been identified as a potential tumor promoter in several cancers. However, the exact mechanism of RNF6 in cancer remains elusive. As in various diseases, RNF6 may be involved in regulating cell growth, cell proliferation, invasion, cell cycle progression, apoptosis and cell adhesion through E3 ligase-mediated ubiquitination. Thus, the research on RNF6 is mainly focused on the ubiquitination of RNF6 in recent years. This article summarizes the role of RNF6 ubiquitination in various physiological and pathological mechanisms, such as Akt/mTOR signaling pathway, Wnt/ß-catenin pathway, RNF6/ERα/Bcl-xL axis, and provides knowledge and understanding for the treatment of diseases.
Assuntos
Carcinogênese/genética , Proteínas de Ligação a DNA/genética , Neoplasias/genética , Transcrição Gênica , Proliferação de Células/genética , Receptor alfa de Estrogênio/genética , Humanos , Neoplasias/patologia , Proteínas Proto-Oncogênicas c-akt/genética , Serina-Treonina Quinases TOR/genética , Ubiquitinação/genética , Proteína bcl-X/genéticaRESUMO
Light intensity influences energy production by increasing photosynthetic carbon, while phosphorus plays an important role in forming the complex nucleic acid structure for the regulation of protein synthesis. These two factors contribute to gene expression, metabolism, and plant growth regulation. In particular, shading is an effective agronomic practice and is widely used to improve the quality of green tea. Genotypic differences between tea cultivars have been observed as a metabolic response to phosphorus deficiency. However, little is known about how the phosphorus supply mediates the effect of shading on metabolites and how plant cultivar gene expression affects green tea quality. We elucidated the responses of the green tea cultivar Longjing43 under three light intensity levels and two levels of phosphorus supply based on a metabolomic analysis by GC×GC-TOF/MS (Two-dimensional Gas Chromatography coupled to Time-of-Flight Mass Spectrometry) and UPLC-Q-TOF/MS (Ultra-Performance Liquid Chromatography-Quadrupole-Time of Flight Mass Spectrometry), a targeted analysis by HPLC (High Performance Liquid Chromatography), and a gene expression analysis by qRT-PCR. In young shoots, the phosphorus concentration increased in line with the phosphate supply, and elevated light intensities were positively correlated with catechins, especially with epigallocatechin of Longjing43. Moreover, when the phosphorus concentration was sufficient, total amino acids in young shoots were enhanced by moderate shading which did not occur under phosphorus deprivation. By metabolomic analysis, phenylalanine, tyrosine, and tryptophan biosynthesis (PTT) were enriched due to light and phosphorus effects. Under shaded conditions, SPX2 (Pi transport, stress, sensing, and signaling), SWEET3 (bidirectional sugar transporter), AAP (amino acid permeases), and GSTb (glutathione S-transferase b) shared the same analogous correlations with primary and secondary metabolite pathways. Taken together, phosphorus status is a crucial factor when shading is applied to increase green tea quality.
Assuntos
Camellia sinensis , Camellia sinensis/metabolismo , Fósforo/metabolismo , Chá/metabolismo , Espectrometria de Massas , Metabolômica , Folhas de Planta/metabolismoRESUMO
Albino tea plants (Camellia sinensis) have been reported to possess highly inhibited metabolism of flavonoids compared to regular green tea leaves, which improves the quality of the tea made from these leaves. However, the mechanisms underlying the metabolism of catechins and flavonols in albino tea leaves have not been well elucidated. In this study, we analyzed a time series of leaf samples in the greening process from albino to green in a thermosensitive leaf-color tea mutant using metabolomics and transcriptomics. The total content of polyphenols dramatically decreased, while flavonols (such as rutin) were highly accumulated in albino leaves compared to in green leaves. After treatment with increasing environment temperature, total polyphenols and catechins were increased in albino mutant tea leaves; however, flavonols (especially ortho-dihydroxylated B-rings such as rutin) were decreased. Meanwhile, weighted gene co-expression network analysis of RNA-seq data suggested that the accumulation of flavonols was highly correlated with genes related to reactive oxygen species scavenging. Histochemical localization further demonstrated that this specific accumulation of flavonols might be related to their biological functions in stress tolerance. These findings suggest that the temperature-stimulated accumulation of total polyphenols and catechins in albino mutant tea leaves was highly induced by enhanced photosynthesis and accumulation of its products, while the initial accumulation and temperature inhibition of flavonols in albino mutant tea leaves were associated with metabolism related to oxidative stress. In conclusion, our results indicate that the biosynthesis of flavonoids could be driven by many different factors, including antioxidation and carbon skeleton storage, under favorable and unfavorable circumstances, respectively. This work provides new insights into the drivers of flavonoid biosynthesis in albino tea leaves, which will further help to increase tea quality by improving cultivation measures.
Assuntos
Camellia sinensis , Catequina , Camellia sinensis/química , Carbono/metabolismo , Catequina/química , Flavonoides/análise , Flavonóis/análise , Metabolômica , Folhas de Planta/química , Proteínas de Plantas/metabolismo , Polifenóis/análise , Espécies Reativas de Oxigênio/metabolismo , Rutina/análise , Chá/genética , Chá/metabolismo , TranscriptomaRESUMO
BACKGROUND: The proper growth and development of tea plants requires moderately acidic soils and relatively low calcium levels, and excessive calcium at high pH can damage tea plant roots. To reveal the effects of calcium on the responses of tea plant to three pH levels (3.5, 5.0 and 6.5), a repeated test of two factors was designed. RESULTS: Root growth and elemental analysis indicated that excessive calcium improved the growth of tea roots at low pH conditions, whereas it did not harm the growth of tea roots under normal and high pH conditions, especially at pH 6.5. Excessive calcium antagonized the absorption and utilization of magnesium by tea plants. Gas chromatography-mass spectrometry results showed that the addition of Ca2+ resulted in the primary metabolism in roots being more active at a low pH level. By contrast, it had obvious adverse effects on the accumulation of root metabolites with high calcium treatment at normal or high pH. Differential metabolites identified using ultra-performance liquid chromatography quadrupole time of flight mass spectrometry indicated that flavonoids demonstrated the largest number of changes, and their biosynthesis was partially enriched with excessive calcium at low and high pH conditions, whereas it was down-regulated under normal pH conditions. Kaempferol 3-(2'-rhamnosyl-6'-acetylgalactoside) 7-rhamnoside, quercetin 3-(6'-sinapoylsophorotrioside) and delphinidin 3-(3'-p-coumaroylglucoside) showed the greatest increase. The results of gene expression related to root growth and calcium regulation were consistent with root growth and root metabolism. CONCLUSION: The overall results demonstrated that high Ca concentrations further aggravate the detrimental effects of high pH to tea roots. However, it is interesting that excessive calcium reduced the harm of a low pH on tea root growth to some extent. © 2021 Society of Chemical Industry.
Assuntos
Cálcio/metabolismo , Camellia sinensis/metabolismo , Proteínas de Plantas/metabolismo , Transporte Biológico , Camellia sinensis/genética , Camellia sinensis/crescimento & desenvolvimento , Concentração de Íons de Hidrogênio , Magnésio/metabolismo , Metabolômica , Proteínas de Plantas/genética , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Solo/químicaRESUMO
BACKGROUND: Greater proportions of purple tea buds and leaves usually appear in the summer, which seriously affects the color and taste quality of green tea products, yet the metabolism of purple tea shoots in summer remains unclear. Here, the metabolomic profiles and gene expression of related flavonoid metabolic pathways in the purple and normal green shoots of 'Longjing 43', and the quality of green tea made with these two phenotypes, were analyzed and compared. RESULTS: Differential metabolites identified using high-performance liquid chromatography-Orbitrap/mass spectrometry indicated that anthocyanin biosynthesis in purple leaves was enriched, with higher levels of anthocyanidins (delphinidin-hexose-coumaroyl showed the greatest increase), proanthocyanidins (oligomers of catechins) and kaempferol glycoside. Expression patterns of the genes ANR, ANS, FLS, LAR, C4H, PAL, CHI, CHS and DFR revealed that the metabolism of anthocyanin is positively regulated by high temperature and/or light levels in summer. Gas chromatography-mass spectrometry results showed that, in purple tea shoots, the metabolism of carbohydrates was enriched whereas that of amino acids was diminished, while their mannose, fructose, d-galactose, sorbose and d-glucose contents were more than double those found in green leaves. A sensory evaluation confirmed that a greater quantity of purple shoots had a greater negative impact on green tea quality because of a bitter taste and dark color (leaves and infusions were tested). CONCLUSIONS: These results highlight the need for and possibility of improving commercial tea quality via cultivation that controls the temperature or light of tea gardens during the summer. © 2019 Society of Chemical Industry.
Assuntos
Antocianinas/biossíntese , Camellia sinensis/metabolismo , Brotos de Planta/metabolismo , Camellia sinensis/química , Camellia sinensis/genética , Camellia sinensis/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Folhas de Planta/química , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Brotos de Planta/química , Brotos de Planta/genética , Brotos de Planta/crescimento & desenvolvimento , Estações do AnoRESUMO
BACKGROUND: Nitrogen (N) nutrition significantly affected metabolism and accumulation of quality-related compounds in tea plant (Camellia sinensis L.). Little is known about the physiological and molecular mechanisms underlying the effects of short-term repression of N metabolism on tea roots and leaves for a short time. RESULTS: In this study, we subjected tea plants to a specific inhibitor of glutamine synthetase (GS), methionine sulfoximine (MSX), for a short time (30 min) and investigated the effect of the inhibition of N metabolism on the transcriptome and metabolome of quality-related compounds. Our results showed that GS activities in tea roots and leaves were significantly inhibited upon MSX treatment, and both tissue types showed a sensitive metabolic response to GS inhibition. In tea leaves, the hydrolysis of theanine decreased with the increase in theanine and free ammonium content. The biosynthesis of all other amino acids was repressed, and the content of N-containing lipids declined, suggesting that short-term inhibition of GS reduces the level of N reutilization in tea leaves. Metabolites related to glycolysis and the tricarboxylic acid (TCA) cycle accumulated after GS repression, whereas the content of amino acids such as glycine, serine, isoleucine, threonine, leucine, and valine declined in the MXS treated group. We speculate that the biosynthesis of amino acids is affected by glycolysis and the TCA cycle in a feedback loop. CONCLUSIONS: Overall, our data suggest that GS repression in tea plant leads to the reprogramming of amino acid and lipid metabolic pathways.
Assuntos
Aminoácidos/metabolismo , Camellia sinensis/metabolismo , Glutamato-Amônia Ligase/antagonistas & inibidores , Metabolismo dos Lipídeos , Metionina Sulfoximina/farmacologia , Proteínas de Plantas/antagonistas & inibidores , Camellia sinensis/efeitos dos fármacos , Camellia sinensis/enzimologia , Metabolismo dos Lipídeos/efeitos dos fármacos , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/metabolismoRESUMO
A series of waterborne polyurethanes (WPU) with crosslinked siloxane were obtained through introducing 3-(2-aminoethylamino)propyldimethoxymethylsilane (APTS) into WPU by in situ polymerization. The properties of WPU modified by APTS were studied through a variety of experimental methods. The water contact angle of the WPU coating surface increased from 64° to 86°, and the water resistance reduced to 3.90% when 3 wt% APTS was added, which improved the coating surface hydrophobicity. Firstly, Fourier transform infrared (FT-IR) and 1H-NMR spectra demonstrated the successful incorporation of APTS to polyurethanes and completed the hydrolytic condensation reaction-generated Si-O-Si crosslinking structure. Furthermore, the surface energy of the membrane was reduced when the crosslinking structure migrated and enriched on the surface of film. Besides, the crosslinking structure was abundant, and the distribution of siloxane in WPU was more uniform.
Assuntos
Materiais Revestidos Biocompatíveis/química , Interações Hidrofóbicas e Hidrofílicas , Poliuretanos/química , Emulsões , Espectroscopia de Ressonância Magnética , Peso Molecular , Tamanho da Partícula , Poliuretanos/síntese química , Espectroscopia de Infravermelho com Transformada de Fourier , TermogravimetriaRESUMO
To uncover mechanism of highly weakened carbon metabolism in chlorotic tea (Camellia sinensis) plants, iTRAQ (isobaric tags for relative and absolute quantification)-based proteomic analyses were employed to study the differences in protein expression profiles in chlorophyll-deficient and normal green leaves in the tea plant cultivar "Huangjinya". A total of 2110 proteins were identified in "Huangjinya", and 173 proteins showed differential accumulations between the chlorotic and normal green leaves. Of these, 19 proteins were correlated with RNA expression levels, based on integrated analyses of the transcriptome and proteome. Moreover, the results of our analysis of differentially expressed proteins suggested that primary carbon metabolism (i.e., carbohydrate synthesis and transport) was inhibited in chlorotic tea leaves. The differentially expressed genes and proteins combined with photosynthetic phenotypic data indicated that 4-coumarate-CoA ligase (4CL) showed a major effect on repressing flavonoid metabolism, and abnormal developmental chloroplast inhibited the accumulation of chlorophyll and flavonoids because few carbon skeletons were provided as a result of a weakened primary carbon metabolism. Additionally, a positive feedback mechanism was verified at the protein level (Mg chelatase and chlorophyll b reductase) in the chlorophyll biosynthetic pathway, which might effectively promote the accumulation of chlorophyll b in response to the demand for this pigment in the cells of chlorotic tea leaves in weakened carbon metabolism.
Assuntos
Camellia sinensis/metabolismo , Carbono/metabolismo , Folhas de Planta/metabolismo , Proteômica/métodos , Regulação da Expressão Gênica de PlantasRESUMO
The qualities of tea (Camellia sinensis) are not clearly understood in terms of integrated leading molecular regulatory network mechanisms behind inorganic phosphate (Pi) limitation. Thus, the present work aims to elucidate transcription factor-dependent responses of quality-related metabolites and the expression of genes to phosphate (P) starvation. The tea plant organs were subjected to metabolomics analysis by GC×GC-TOF/MS and UPLC-Q-TOF/MS along with transcription factors and 13 metabolic genes by qRT-PCR. We found P starvation upregulated SPX2 and the change response of Pi is highly dependent on young shoots. This led to increased change in abundance of carbohydrates (fructose and glucose), amino acids in leaves (threonine and methionine), and root (phenylalanine, alanine, tryptophan, and tyrosine). Flavonoids and their glycosides accumulated in leaves and root exposed to P limitation was consistent with the upregulated expression of anthocyanidin reductase (EC 1.3.1.77), leucoanthocyanidin dioxygenase (EC 1.4.11.19) and glycosyltransferases (UGT78D1, UGT78D2 and UGT57L12). Despite the similar kinetics and high correlation response of Pi and SPX2 in young shoots, predominating theanine and other amino acids (serine, threonine, glutamate, valine, methionine, phenylalanine) and catechin (EGC, EGCG and CG) content displayed opposite changes in response to Pi limitation between Fengqing and Longjing-43 tea cultivars.
Assuntos
Aminoácidos/metabolismo , Camellia sinensis/metabolismo , Flavonoides/metabolismo , Fosfatos/deficiência , Camellia sinensis/genética , Glicosiltransferases/genética , Glicosiltransferases/metabolismo , Oxigenases/genética , Oxigenases/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estresse FisiológicoRESUMO
BACKGROUND: As the predominant secondary metabolic pathway in tea plants, flavonoid biosynthesis increases with increasing temperature and illumination. However, the concentration of most flavonoids decreases greatly in light-sensitive tea leaves when they are exposed to light, which further improves tea quality. To reveal the metabolism and potential functions of flavonoids in tea leaves, a natural light-sensitive tea mutant (Huangjinya) cultivated under different light conditions was subjected to metabolomics analysis. RESULTS: The results showed that chlorotic tea leaves accumulated large amounts of flavonoids with ortho-dihydroxylated B-rings (e.g., catechin gallate, quercetin and its glycosides etc.), whereas total flavonoids (e.g., myricetrin glycoside, epigallocatechin gallate etc.) were considerably reduced, suggesting that the flavonoid components generated from different metabolic branches played different roles in tea leaves. Furthermore, the intracellular localization of flavonoids and the expression pattern of genes involved in secondary metabolic pathways indicate a potential photoprotective function of dihydroxylated flavonoids in light-sensitive tea leaves. CONCLUSIONS: Our results suggest that reactive oxygen species (ROS) scavenging and the antioxidation effects of flavonoids help chlorotic tea plants survive under high light stress, providing new evidence to clarify the functional roles of flavonoids, which accumulate to high levels in tea plants. Moreover, flavonoids with ortho-dihydroxylated B-rings played a greater role in photo-protection to improve the acclimatization of tea plants.
Assuntos
Camellia sinensis/metabolismo , Flavonoides/metabolismo , Luz , Metabolômica , Folhas de Planta/metabolismo , Camellia sinensis/efeitos da radiação , Catequina/análogos & derivados , Catequina/metabolismo , Glicosídeos/metabolismo , Redes e Vias Metabólicas/efeitos da radiação , Espécies Reativas de Oxigênio/metabolismoRESUMO
BACKGROUND: Nitrogen (N) plays an important role in the formation of tea quality-related compounds, like amino acids and flavor/aroma origin compounds. Lipids, which have been reported to be affected by N deficiency, are precursors to the generation of flavor/aroma origin compounds in tea plant. However, there is no literature about the lipid profiles of tea plant affected by N fertilization. Hence, we hypothesize that the biosynthesis of flavor-related compounds in tea was affected by N through its regulation of lipid metabolism. RESULTS: In this study, mature leaves and new shoots of tea plant grown under three N levels at the rates of 0, 285 and 474 kg/ha were applied for ultra-performance liquid chromatography-mass spectrometry (UPLC/MS) based lipidomic analysis. Totally, 178 lipid species were identified. The results showed that the composition of lipid compounds in mature leaves and new shoots varied dramatically, which was also affected by N levels. The higher content of the storage lipid TAG and higher carbon (C)/N ratio in mature leaves than that of new shoots in tea plants grown under low N level (0 kg/ha) suggested that tea plants could remobilize the C stored in TAG to maintain their C/N balance and help to improve the quality of tea. N fertilization resulted in a higher content of the compounds 36:6 MGDG and 36:6 DGDG. Since these compounds contain linolenic acid (18:3), a precursor to the formation of aroma origin compounds, we suggested their increase could contribute to the quality of tea. CONCLUSIONS: Taken together, the present work indicated that appropriate application of N fertilizer could balance the lipid metabolism and the formation of flavor/aroma origin compounds, which help to improve the quality of tea. Moreover, excess N fertilization might deteriorate the aroma quality of made tea due to increases of precursors leading to grassy odor.
Assuntos
Camellia sinensis/metabolismo , Metabolismo dos Lipídeos , Nitrogênio/metabolismo , Cromatografia Líquida de Alta Pressão , Fertilizantes , Folhas de Planta/metabolismo , Brotos de Planta/metabolismoRESUMO
Ephrin-A2, a member of the Eph/ephrin family, is associated with tumorigenesis and tumor progression. This study aimed to assess the diagnostic and prognostic value of both serum and tissue levels of Ephrin-A2 in prostate cancer (PCa) management. One hundred and forty-five frozen prostate tissues, 55 paraffin-embedded prostate tissues, 88 serum samples, and seven prostate cell lines (RWPE-1, LNCaP, LNCaP-LN3, PC-3, PC-3M, PC-3M-LN4, and DU145) were examined via quantitative reverse transcription-PCR (qRT-PCR), immunohistochemistry, enzyme-linked immunosorbent assay, and western blotting. Induced Ephrin-A2 messenger RNA (mRNA) or protein expression was detected in 8.6 % (5/58) benign prostatic hyperplasia (BPH), 59.8 % (52/87) PCa, and five prostate cancer cell lines. Ephrin-A2 immunostaining was present in 6.7 % (1/15) patients with BPHs and 62.5 % (25/40) clinically localized PCa. Accordingly, serum Ephrin-A2 was significantly higher in PCa patients compared to those in the BPH patients and controls (P < 0.001). The expression of Ephrin-A2 was higher in tumor patients with an elevated Gleason score or T3-T4 staging. Ephrin-A2 expression was correlated with Ki-67 expression in PCa patients, both at the gene scale and protein level. Our data indicate that Ephrin-A2 is a potential diagnostic and prognostic biomarker and a promising molecular therapeutic target to attenuate prostate cancer progression.
Assuntos
Biomarcadores Tumorais/biossíntese , Efrina-A2/biossíntese , Prognóstico , Neoplasias da Próstata/genética , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/sangue , Biomarcadores Tumorais/genética , Efrina-A2/sangue , Efrina-A2/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Antígeno Ki-67/biossíntese , Masculino , Gradação de Tumores , Estadiamento de Neoplasias , Células Neoplásicas Circulantes/patologia , Próstata/patologia , Neoplasias da Próstata/sangue , Neoplasias da Próstata/patologiaRESUMO
Sleep stage scoring is a hotspot in the field of medicine and neuroscience.Visual inspection of sleep is laborious and the results may be subjective to different clinicians.Automatic sleep stage classification algorithm can be used to reduce the manual workload.However,there are still limitations when it encounters complicated and changeable clinical cases.The purpose of this paper is to develop an automatic sleep staging algorithm based on the characteristics of actual sleep data.In the proposed improved K-means clustering algorithm,points were selected as the initial centers by using a concept of density to avoid the randomness of the original K-means algorithm.Meanwhile,the cluster centers were updated according to the'Three-Sigma Rule'during the iteration to abate the influence of the outliers.The proposed method was tested and analyzed on the overnight sleep data of the healthy persons and patients with sleep disorders after continuous positive airway pressure(CPAP)treatment.The automatic sleep stage classification results were compared with the visual inspection by qualified clinicians and the averaged accuracy reached 76%.With the analysis of morphological diversity of sleep data,it was proved that the proposed improved K-means algorithm was feasible and valid for clinical practice.
Assuntos
Algoritmos , Processamento de Sinais Assistido por Computador , Fases do Sono , Transtornos do Sono-Vigília/diagnóstico , Análise por Conglomerados , Pressão Positiva Contínua nas Vias Aéreas , Eletroencefalografia , Humanos , PolissonografiaRESUMO
The strategy of rational design to engineer enzymes is to predict the potential mutants based on the understanding of the relationships between protein structure and function, and subsequently introduce the mutations using the site-directed mutagenesis. Rational design methods are universal, relatively fast and have the potential to be developed into algorithms that can quantitatively predict the performance of the designed sequences. Compared to the protein stability, it was more challenging to design an enzyme with improved activity or selectivity, due to the complexity of enzyme molecular structure and inadequate understanding of the relationships between enzyme structures and functions. However, with the development of computational force, advanced algorithm and a deeper understanding of enzyme catalytic mechanisms, rational design could significantly simplify the process of engineering enzyme functions and the number of studies applying rational design strategy has been increasing. Here, we reviewed the recent advances of applying the rational design strategy to engineer enzyme functions including activity and enantioselectivity. Five strategies including multiple sequence alignment, strategy based on steric hindrance, strategy based on remodeling interaction network, strategy based on dynamics modification and computational protein design are discussed and the successful cases using these strategies are introduced.