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1.
Plant Cell Environ ; 47(7): 2693-2709, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38576334

RESUMO

As a well-conserved histone variant, H2A.Z epigenetically regulates plant growth and development as well as the interaction with environmental factors. However, the role of H2A.Z in response to salt stress remains unclear, and whether nucleosomal H2A.Z occupancy work on the gene responsiveness upon salinity is obscure. Here, we elucidate the involvement of H2A.Z in salt response by analysing H2A.Z disorder plants with impaired or overloaded H2A.Z deposition. The salt tolerance is dramatically accompanied by H2A.Z deficiency and reacquired in H2A.Z OE lines. H2A.Z disorder changes the expression profiles of large-scale of salt responsive genes, announcing that H2A.Z is required for plant salt response. Genome-wide H2A.Z mapping shows that H2A.Z level is induced by salt condition across promoter, transcriptional start site (TSS) and transcription ending sites (-1 kb to +1 kb), the peaks preferentially enrich at promoter regions near TSS. We further show that H2A.Z deposition within TSS provides a direct role on transcriptional control, which has both repressive and activating effects, while it is found generally H2A.Z enrichment negatively correlate with gene expression level response to salt stress. This study shed light on the H2A.Z function in salt tolerance, highlighting the complex regulatory mechanisms of H2A.Z on transcriptional activity for yielding appropriate responses to particularly environmental stress.


Assuntos
Arabidopsis , Regulação da Expressão Gênica de Plantas , Histonas , Histonas/metabolismo , Arabidopsis/genética , Arabidopsis/fisiologia , Transcrição Gênica/efeitos dos fármacos , Estresse Salino/genética , Tolerância ao Sal/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Regiões Promotoras Genéticas/genética , Nucleossomos/metabolismo
2.
Physiol Plant ; 176(4): e14411, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38973028

RESUMO

Plant growth-promoting rhizobacteria (PGPR) are known for their role in ameliorating plant stress, including alkaline stress, yet the mechanisms involved are not fully understood. This study investigates the impact of various inoculum doses of Bacillus licheniformis Jrh14-10 on Arabidopsis growth under alkaline stress and explores the underlying mechanisms of tolerance enhancement. We found that all tested doses improved the growth of NaHCO3-treated seedlings, with 109 cfu/mL being the most effective. Transcriptome analysis indicated downregulation of ethylene-related genes and an upregulation of polyamine biosynthesis genes following Jrh14-10 treatment under alkaline conditions. Further qRT-PCR analysis confirmed the suppression of ethylene biosynthesis and signaling genes, alongside the activation of polyamine biosynthesis genes in NaHCO3-stressed seedlings treated with Jrh14-10. Genetic analysis showed that ethylene signaling-deficient mutants (etr1-3 and ein3-1) exhibited greater tolerance to NaHCO3 than the wild type, and the growth-promoting effect of Jrh14-10 was significantly diminished in these mutants. Additionally, Jrh14-10 was found unable to produce 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase, indicating it does not reduce the ethylene precursor ACC in Arabidopsis. However, Jrh14-10 treatment increased the levels of polyamines (putrescine, spermidine, and spermine) in stressed seedlings, with spermidine particularly effective in reducing H2O2 levels and enhancing Fv/Fm under NaHCO3 stress. These findings reveal a novel mechanism of PGPR-induced alkaline tolerance, highlighting the crosstalk between ethylene and polyamine pathways, and suggest a strategic redirection of S-adenosylmethionine towards polyamine biosynthesis to combat alkaline stress.


Assuntos
Arabidopsis , Bacillus licheniformis , Etilenos , Poliaminas , Arabidopsis/genética , Arabidopsis/efeitos dos fármacos , Arabidopsis/metabolismo , Arabidopsis/microbiologia , Arabidopsis/fisiologia , Etilenos/metabolismo , Poliaminas/metabolismo , Bacillus licheniformis/metabolismo , Bacillus licheniformis/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Estresse Fisiológico , Plântula/efeitos dos fármacos , Plântula/genética , Plântula/fisiologia , Plântula/metabolismo , Álcalis/farmacologia , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética
3.
Plant Cell Rep ; 43(2): 33, 2024 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-38200226

RESUMO

KEY MESSAGE: The study on melatonin biosynthesis mutant snat1snat2 revealed that endogenous melatonin plays an important role in salt responsiveness by mediating auxin signaling. Melatonin is a pleiotropic signaling molecule, which, besides being involved in multiple growth and developmental processes, also mediates environmental stress responses. However, whether and how endogenous melatonin is involved in salt response has not been determined. In this study, we elucidated the involvement of endogenous melatonin in salt response by investigatiing the impact of salt stress on a double mutant of Arabidopsis (snat1snat2) defective in melatonin biosynthesis genes SNAT1 and SNAT2. This mutant was found to exhibit salt sensitivity, manifested by unhealthy growth, ion imbalance and ROS accumulation under salt stress. Transcriptomic profiles of snat1snat2 revealed that the expression of a large number of salt-responsive genes was affected by SNAT defect, and these genes were closely related to the synthesis of auxin and several signaling pathways. In addition, the salt-sensitive growth phenotype of snat1snat2 was alleviated by the application of exogenous auxin. Our results show that endogenous melatonin may be essential for plant salt tolerance, a function that could be correlated with diverse activity in mediating auxin signaling.


Assuntos
Arabidopsis , Melatonina , Arabidopsis/genética , Ácidos Indolacéticos , Fenótipo , Estresse Salino/genética
4.
Proc Natl Acad Sci U S A ; 118(26)2021 06 29.
Artigo em Inglês | MEDLINE | ID: mdl-34155099

RESUMO

Multiple independent sequence variants of the hTERT locus have been associated with telomere length and cancer risks in genome-wide association studies. Here, we identified an intronic variable number tandem repeat, VNTR2-1, as an enhancer-like element, which activated hTERT transcription in a cell in a chromatin-dependent manner. VNTR2-1, consisting of 42-bp repeats with an array of enhancer boxes, cooperated with the proximal promoter in the regulation of hTERT transcription by basic helix-loop-helix transcription factors and maintained hTERT expression during embryonic stem-cell differentiation. Genomic deletion of VNTR2-1 in MelJuSo melanoma cells markedly reduced hTERT transcription, leading to telomere shortening, cellular senescence, and impairment of xenograft tumor growth. Interestingly, VNTR2-1 lengths varied widely in human populations; hTERT alleles with shorter VNTR2-1 were underrepresented in African American centenarians, indicating its role in human aging. Therefore, this polymorphic element is likely a missing link in the telomerase regulatory network and a molecular basis for genetic diversities of telomere homeostasis and age-related disease susceptibilities.


Assuntos
Repetições Minissatélites/genética , Polimorfismo Genético , Telomerase/genética , Ativação Transcricional , Negro ou Afro-Americano/genética , Idoso de 80 Anos ou mais , Animais , Sequência de Bases , Diferenciação Celular/genética , Linhagem Celular , Proliferação de Células/genética , Cromossomos Artificiais Bacterianos/genética , Elementos E-Box/genética , Genoma Humano , Células-Tronco Embrionárias Humanas/metabolismo , Humanos , Camundongos Nus , Neoplasias/genética , Neoplasias/patologia , Regiões Promotoras Genéticas , Ligação Proteica/genética , Deleção de Sequência/genética , Homeostase do Telômero/genética
5.
Int J Mol Sci ; 25(1)2024 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-38203741

RESUMO

Male gametophyte development in plants relies on the functions of numerous genes, whose expression is regulated by transcription factors (TFs), non-coding RNAs, hormones, and diverse environmental stresses. Several excellent reviews are available that address the genes and enzymes associated with male gametophyte development, especially pollen wall formation. Growing evidence from genetic studies, transcriptome analysis, and gene-by-gene studies suggests that TFs coordinate with epigenetic machinery to regulate the expression of these genes and enzymes for the sequential male gametophyte development. However, very little summarization has been performed to comprehensively review their intricate regulatory roles and discuss their downstream targets and upstream regulators in this unique process. In the present review, we highlight the research progress on the regulatory roles of TF families in the male gametophyte development of flowering plants. The transcriptional regulation, epigenetic control, and other regulators of TFs involved in male gametophyte development are also addressed.


Assuntos
Magnoliopsida , Fatores de Transcrição , Humanos , Fatores de Transcrição/genética , Epigenômica , Perfilação da Expressão Gênica , Pólen/genética
6.
Plant Dis ; 2023 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-37415356

RESUMO

Broccoli (Brassica oleracea var. italica) is not only an important crop worldwide with a large amount of production and consumption annually, but also rich in biologically active compounds (Surh et al., 2021). In November 2022, an unknown leaf blight was observed in the Broccoli planting area, Wenzhou City of Zhejiang Province (28.05 °N, 120.31 °E). Symptoms initially occurred at the leaf margin with yellow to gray lesions that were irregular and wilting. Approximately 10% of the surveyed plants were affected. To determine the pathogen, leaves with blight were collected randomly from five B. oleracea plants. Tissue blocks (3×3 mm) from diseased leaf portions were disinfected with 75% ethanol, rinsed three times with sterilized water, placed aseptically onto potato dextrose agar (PDA) medium, and incubated for 5 days at 28℃ in darkness. Seven fungal isolates with the same morphology were obtained using the spore method. The observed colonies were circular, taupe, pewter in color with light gray edging and many cottony aerial mycelia. Conidia were straight, curved or slightly bent, ellipsoidal to fusiform, and septate (typically 4-8 septa per conidium), with the size of 50.0-90.0 µm × 10.0-20.0 µm (n=30). The conidia had a slightly protruding and truncate hilum. These morphological characteristics were consistent with Exserohilum rostratum (Sharma et al., 2014). To further identify the pathogen, isolate WZU-XLH1 was chosen as a representative and the internal transcribed spacer (ITS) and glyceraldehyde-3-phosphate dehydrogenase-like (GAPDH) gene were amplified and sequenced using primer pairs ITS1/ITS4 (White et al., 1990) and Gpd1/Gpd2 (Berbee et al., 1999), respectively. The ITS and gpd gene sequences of isolate WZU-XLH1 were deposited in the GenBank database with accession numbers OQ750113 and OQ714500, respectively. BLASTn analysis showed matches of 568/571 (MH859108) and 547/547 (LT882549) with Exserohilum rostratum CBS 188.68. A neighbor-joining phylogenetic tree was constructed by combining the two sequenced loci, this isolate in the E. rostratum species complex clade at 71% bootstrap support.To verify the pathogenicity of the isolate, ten healthy Broccoli (cultivar 'You Xiu') seedlings with at least five leaves were divided into two groups: one group was inoculated with the isolate, while the other group served as a control. After surface disinfection with 75% ethanol and wiping with sterile water, tiny wounds were made on two leaves (two wounds in one leaf) using an inoculation needle. Fungal culture plugs cut from the isolate were placed on the wounds, while sterile PDA plugs served as the control. The leaves were sealed in wet airtight bags to retain moisture at room temperature with natural light (Cao et al., 2022). After five days, all leaves inoculated with isolate WZU-XLH1 showed symptoms identical to those observed in the field, with no symptoms present in the control group. The pathogenicity was confirmed by repeating the test in triplicate, and fungi re-isolated from symptomatic leaves were identified as E. rostratum by the morphological and molecular methods described above. To the best of our knowledge, this is the first report of E. rostratum causing leaf blight on broccoli in China. This study contributes to our understanding of B. oleracea leaf blight and establishes a basis for future studies on E. rostratum to develop management strategies.

7.
Int J Mol Sci ; 24(14)2023 Jul 19.
Artigo em Inglês | MEDLINE | ID: mdl-37511417

RESUMO

Salt is one of the most important environmental factors in crop growth and development. N6-methyladenosine (m6A) is an epigenetic modification that regulates plant-environment interaction at transcriptional and translational levels. Sugar beet is a salt-tolerant sugar-yielding crop, but how m6A modification affects its response to salt stress remains unknown. In this study, m6A-seq was used to explore the role of m6A modification in response to salt stress in sugar beet (Beta vulgaris). Transcriptome-wide m6A methylation profiles and physiological responses to high salinity were investigated in beet roots. After treatment with 300 mM NaCl, the activities of peroxidase and catalase, the root activity, and the contents of Na+, K+, and Ca2+ in the roots were significantly affected by salt stress. Compared with the control plants, 6904 differentially expressed genes (DEGs) and 566 differentially methylated peaks (DMPs) were identified. Association analysis revealed that 243 DEGs contained DMP, and 80% of these DEGs had expression patterns that were negatively correlated with the extent of m6A modification. Further analysis verified that m6A methylation may regulate the expression of some genes by controlling their mRNA stability. Functional analysis revealed that m6A modifications primarily affect the expression of genes involved in energy metabolism, transport, signal transduction, transcription factors, and cell wall organization. This study provides evidence that a post-transcriptional regulatory mechanism mediates gene expression during salt stress by affecting the stability of mRNA in the root.


Assuntos
Beta vulgaris , Beta vulgaris/metabolismo , Epigenoma , Estresse Salino/genética , Transcriptoma , Açúcares/metabolismo , Regulação da Expressão Gênica de Plantas , Raízes de Plantas/metabolismo , Estresse Fisiológico/genética
8.
Int J Mol Sci ; 24(19)2023 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-37833966

RESUMO

Salt stress severely restricts plant growth and crop production, which is accompanied by accumulation of reactive oxygen species (ROS) that disturb cell redox homeostasis and oxidize redox-sensitive proteins. Eutrema salsugineum, a halophytic species closely related to Arabidopsis, shows a high level of tolerance to salinity and is increasingly used as a model plant in abiotic stress biology. To understand redox modifications and signaling pathways under salt stress, we used tandem mass tag (TMT)-based proteomics to quantify the salt-induced changes in protein redox modifications in E. salsugineum. Salt stress led to increased oxidative modification levels of 159 cysteine sites in 107 proteins, which play roles in carbohydrate and energy metabolism, transport, ROS homeostasis, cellular structure modulation, and folding and assembly. These lists of unknown redox reactive proteins in salt mustard lay the foundation for future research to understand the molecular mechanism of plant salt response. However, glutathione peroxidase (GPX) is one of the most important antioxidant enzymes in plants. Our research indicates that EsGPX may be involved in regulating ROS levels and that plants with overexpressed EsGPX have much improved salt tolerance.


Assuntos
Arabidopsis , Brassicaceae , Tolerância ao Sal , Espécies Reativas de Oxigênio/metabolismo , Proteômica , Proteínas de Plantas/genética , Brassicaceae/metabolismo , Arabidopsis/metabolismo , Oxirredução , Regulação da Expressão Gênica de Plantas
9.
Plant Cell Environ ; 43(6): 1452-1466, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32022936

RESUMO

Nuclear pore complexes (NPCs) are main channels controlling nucleocytoplasmic transport and are composed of approximately 30 nucleoporins (NUPs). Emerging evidence suggests that some NUP genes have specialized functions that challenge the traditional view of NPCs as structures of uniform composition. Here, we analysed the role of six outer-ring components of NPC at normal and warm growth temperatures by examining their loss-of-function mutants in Arabidopsis thaliana. All six NUP subunits, NUP85, NUP96, NUP 133, NUP 160, SEH1 and HOS1, have a non-redundant temperature-influenced function in one or more of the processes, including rosette growth, leaf architecture and intracellular immune receptor-mediated disease resistance. At the molecular level, NUP85 and NUP133 are required for mRNA export only at warm temperature and play a larger role in the localization of transcription factor at warm temperature. In addition, NUP96 and HOS1 are essential for the expression of high temperature-responsive genes, which is correlated with their larger activity in facilitating nuclear accumulation of the transcription factor PIF4 at warm temperature. Our results show that subunits of NPC have differential roles at different temperatures, suggesting the existence of temperature-influenced NPC complexes and activities.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/imunologia , Complexo de Proteínas Formadoras de Poros Nucleares/metabolismo , Desenvolvimento Vegetal , Imunidade Vegetal , Temperatura , Arabidopsis/genética , Arabidopsis/microbiologia , Núcleo Celular/metabolismo , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Mutação com Perda de Função , Fenótipo , Transporte de RNA/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Frações Subcelulares/metabolismo , Transcrição Gênica , Virulência
10.
Lipids Health Dis ; 18(1): 56, 2019 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-30832728

RESUMO

BACKGROUND: ω-3 polyunsaturated fatty acids (PUFAs) are synthesized from α-Linolenic acid (ALA, C18:3ω3) and play important roles in anti-inflammatory and antioxidant responses in mammal cells. ALA is an essential fatty acid which cannot be produced within the human body and must be acquired through diet. The purpose of this study was to evaluate the potential of a novel microalgal strain (HDMA-20) as a source of ω-3 PUFAs including ALA and eicosatetraenoic acid (ETA, C20:4ω3). METHOD: Phylogenetic Neighbor-Joining analysis based on 18S ribosomal DNA sequence was used to identify the microalga strain HDMA-20. Autotrophic condition was chosen to cultivate HDMA-20 to reduce the cultivation cost. GC-MS was used to determine the fatty acid composition of HDMA-20 lipid. RESULTS: A microalgal strain (HDMA-20) from Lake Chengfeng (Daqing, Heilongjiang province, China) was found to accumulate high content of ω-3 PUFAs (63.4% of total lipid), with ALA and eicosatetraenoic acid (ETA, C20:4ω3) accounting for 35.4 and 9.6% of total lipid, respectively. Phylogenetic analysis based on 18S ribosomal DNA sequences suggested that the HDMA-20 belonged to genus Monoraphidium (Selenastraceae, Sphaeropleales) and its 18S rDNA sequence information turned out to be new molecular record of Monoraphidium species. The biomass productivity and lipid content of HDMA-20 were also investigated under autotrophic condition. The biomass productivity of HDMA-20 reached 36.3 mg L- 1 day- 1, and the lipid contents was 22.6% of dry weight. CONCLUSION: HDMA-20 not only represent an additional source of ALA, but also a totally new source of ETA. The high content of ω-3 PUFAs, especially ALA, of HDMA-20, makes it suitable as a source of nutrition supplements for human health. In addition, HDMA-20 exhibited good properties in growth and lipid accumulation, implying its potential for cost-effective ω-3 PUFAs production in future.


Assuntos
Ácidos Araquidônicos/isolamento & purificação , Clorofíceas/metabolismo , Suplementos Nutricionais/análise , Microalgas/metabolismo , Ácido alfa-Linolênico/isolamento & purificação , Ácidos Araquidônicos/biossíntese , Processos Autotróficos/fisiologia , Biomassa , China , Clorofíceas/classificação , Clorofíceas/genética , Clorofíceas/crescimento & desenvolvimento , Suplementos Nutricionais/provisão & distribuição , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Lagos , Metaboloma/fisiologia , Microalgas/classificação , Microalgas/genética , Microalgas/crescimento & desenvolvimento , Filogenia , RNA Ribossômico 18S/genética , Ácido alfa-Linolênico/biossíntese
11.
Ecotoxicol Environ Saf ; 171: 12-25, 2019 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-30593996

RESUMO

Soil salinity is a major abiotic stress affecting crop growth and productivity. Ricinus communis has good salt tolerance and is also an important oilseed crop throughout the world. Early seedling stage (such as cotyledon expansion stage) is the most vulnerable period for plant under stresses. However, little information exist concerning the physiological and molecular mechanisms of Ricinus communis seedlings and the role play by cotyledons and true leaf under salt stress. In the present study, biomass, photosynthesis, chlorophyll fluorescence, inorganic ions and organic solutes contents were measured, and two dimensional gel electrophoresis-based proteomic technology was employed to identify the differentially abundant proteins in the salt-treated Ricinus communis cotyledons and true leaves. The results showed that salt stress reduced growth and photosynthesis in the seedlings. With increasing salinity, the Na+ content increased and K+ content decreased in both cotyledons and leaves, but the true leaves had lower Na+ and higher K+ contents. Soluble sugars and proline are the primary organic solutes to cope with osmotic stress. In addition, proteomic analysis revealed 30 and 42 differentially accumulated protein spots in castor cotyledon and true leaf under salt stress, respectively. Most of the identified proteins were involved in carbohydrate and energy metabolism, photosynthesis, genetic information process, reactive oxygen species metabolism, amino acid metabolism and cell structure. The physiological and proteomic results highlighted that cotyledons accumulated a large number of Na+ and provided more energy to help true leaves cope with salt stress. The true leaves saved carbon structures to synthesize osmotic substances, and the enhancement of chlorophyll synthesis and electron transfer in true leaves could also maintain photosynthesis under salt stress. These findings provide new insights into different physiological mechanisms in cotyledon and true leaf of Ricinus communis response to salt stress during early seedling stage.


Assuntos
Cotilédone/metabolismo , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Ricinus , Salinidade , Tolerância ao Sal , Plântula/metabolismo , Biomassa , Metabolismo Energético , Pressão Osmótica/fisiologia , Fotossíntese , Potássio/metabolismo , Prolina/metabolismo , Proteômica , Plântula/crescimento & desenvolvimento , Sódio/metabolismo , Cloreto de Sódio/análise
12.
Planta ; 247(3): 715-732, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29185033

RESUMO

MAIN CONCLUSION: Ribosome activation and sugar metabolic process mainly act on the regulation of salt tolerance in the bioenergy crop Helianthus tuberosus L. as dissected by integrated transcriptomic and proteomic analyses. Helianthus tuberosus L. is an important halophyte plant that can survive in saline-alkali soil. It is vitally necessary to build an available genomic resource to investigate the molecular mechanisms underlying salt tolerance in H. tuberosus. De novo assembly and annotation of transcriptomes were built for H. tuberosus using a HiSeq 4000 platform. 293,823 transcripts were identified and annotated into 190,567 unigenes. In addition, iTRAQ-labeled quantitative proteomics was carried out to detect global protein profiling as a response to salt stress. Comparative omics analysis showed that 5432 genes and 43 proteins were differentially expressed in H. tuberosus under salt stress, which were enriched in the following processes: carbohydrate metabolism, ribosome activation and translation, oxidation-reduction and ion binding. The reprogramming of transcript and protein works suggested that the induced activity of ribosome and sugar signaling may endue H. tuberosus with salt tolerance. With high-quality sequencing and annotation, the obtained transcriptomics and proteomics provide a robust genomic resource for dissecting the regulatory molecular mechanism of H. tuberosus in response to salt stress.


Assuntos
Helianthus/metabolismo , Raízes de Plantas/metabolismo , Tolerância ao Sal , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Helianthus/genética , Peróxido de Hidrogênio/metabolismo , Malondialdeído/metabolismo , Raízes de Plantas/fisiologia , Reação em Cadeia da Polimerase , Proteômica , Tolerância ao Sal/genética , Transcriptoma/genética
13.
Biomed Chromatogr ; 32(11): e4336, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30003562

RESUMO

In the present study, a 10-position modified of camptothecin, 10-propionyloxy camptothecin (PCPT) was esterified from 10-hydroxcamptothecin (HCPT), which could metabolize to HCPT in vivo. PCPT displayed a relatively stronger antitumor activity in vitro and in vivo. Thereafter a simple, sensitive and rapid HPLC method coupled with a fluorescence detector was developed and validated for the assay of PCPT and its active metabolite HCPT in rat plasma. The method was validated for accuracy, precision, linearity, selectivity and recovery. The validated method was successfully applied to the pharmacokinetic study of PCPT in rats after intravenous administration. The results showed that PCPT could be mainly converted to HCPT in plasma with the AUC0-∞ value of 3.69 ± 4.44 and 311.16 ± 188.81 ng h/mL for PCPT and HCPT, respectively.


Assuntos
Antineoplásicos , Camptotecina , Animais , Antineoplásicos/sangue , Antineoplásicos/farmacocinética , Antineoplásicos/farmacologia , Camptotecina/análogos & derivados , Camptotecina/sangue , Camptotecina/farmacocinética , Camptotecina/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Modelos Lineares , Masculino , Ratos , Ratos Wistar , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Ensaios Antitumorais Modelo de Xenoenxerto
14.
Molecules ; 23(3)2018 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-29562617

RESUMO

Cancer is one of the most major diseases that threatens human health and life. The aim of this work was to obtain novel anticancer molecules from D. fragrans, a kind of medicinal plant. The structure of the new compound was identified using spectroscopic data (¹H-NMR, 13C-NMR and two dimensions NMR). Its anticancer properties were evaluated using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay against four human cells including lung cancer cells (A549), breast cancer cells (MCF-7), gastric cancer cells (SGC7901) and noncancerous human umbilical vein endothelial cells (HUVEC). A new phenylpropanoid-(E)-caffeic acid-9-O-ß-d-xylpyranosyl-(1→2)-ß-d-glucopyranosyl ester (1), with seven known compounds (2-8)-was isolated. The IC50 value of compound 1 against MCF-7 cells was 2.65 ± 0.14 µM, and the IC50 values of compound 8 against three cancer cells were below 20 µM.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Dryopteris/química , Fenóis/farmacologia , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/isolamento & purificação , Espectroscopia de Ressonância Magnética Nuclear de Carbono-13 , Linhagem Celular Tumoral , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Fenóis/química , Fenóis/isolamento & purificação , Espectroscopia de Prótons por Ressonância Magnética
17.
J Sci Food Agric ; 97(1): 88-94, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26917428

RESUMO

BACKGROUND: Plant cell suspension cultures have emerged as a potential source of secondary metabolites for food additives and pharmaceuticals. In this study inulin accumulation and its degree of polymerisation (DP) in the treated cells in the same medium were investigated after treatment with six types of elicitors. RESULTS: An in vitro cell suspension culture of Jerusalem artichoke (Helianthus tuberosus L.) was optimised by adding an extra nitrogen source. According to the growth kinetics, a maximum biomass of 5.48 g L-1 was obtained from the optimal cell suspension medium consisted of Murashige and Skoog basic medium (MS) + 1.0 mg L-1 α-naphthalene acetic acid (NAA) + 1.0 mg L-1 6-benzylaminopurine (6-BA) + 0.5 mg L-1 proline + 1.0 mg L-1 glutamine. Methyl jasmonate (MeJA, 250 µmol L-1 ) treatment for 15 days led to the highest levels of inulin (2955.27 ± 9.81 mg L-1 compared to control of 1217.46 ± 0.26 mg L-1 ). The elicited effect of five elicitors to the suspension cells of Jerusalem artichoke is as follows: AgNO3 (Ag, 10 µmol L-1 ), salicylic acid (SA, 75 µmol L-1 ), chitosan (KJT, 40 mg L-1 ), Trichoderma viride (Tv, 90 mg L-1 ), yeast extract (YE, 0.25 mg L-1 ), and the corresponding content of inulin is increased by 2.05-, 1.93-, 1.76-, 1.44- and 1.18-fold compared to control, respectively. The obvious effect on the percentage of lower DP in inulin was observed in cells treated with 40 mg L-1 KJT, 0.25 mg L-1 YE and 10 µmol L-1 Ag. CONCLUSIONS: Among the six types of elicitors, the descending order of inulin content is MeJA > Ag > SA > KJT > Tv > YE. For the purpose inulin with lower DP and its application to prebiotic food, three elicitors, including KJT, YE and Ag, can be used for the elicitation. © 2016 Society of Chemical Industry.


Assuntos
Meios de Cultura/química , Helianthus/crescimento & desenvolvimento , Helianthus/metabolismo , Inulina/química , Inulina/metabolismo , Técnicas de Cultura de Células , Células Cultivadas , Meios de Cultura/metabolismo , Polimerização
18.
Xenobiotica ; 46(9): 831-40, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26744836

RESUMO

1. Camptothecin (CPT) and its derivatives are potent candidate compounds in treating cancers. However, their clinical applications are largely restricted by severe toxicities. 2. The solute carrier transporters (SLCs), particularly the organic anion transporting polypeptides and organic anion/cation transporters (OATs/OCTs) are widely expressed in human key organs and responsible for the cellular influx of many substances including endogenous substrates and many clinically important drugs. Drug-drug interactions through SLCs often result in unsatisfied therapeutic outcomes and/or unexpected toxicities. 3. This study investigated the inhibitory effects of CPT and its eight derivatives on the cellular uptake of specific substrates mediated by the essential SLCs in over-expressing Human embryonic kidney 293 cells. 4. Our data revealed that CPT, 10-hydroxycamptothecin (HCPT), 10-methoxycamptothecin (MCPT) and 9-nitrocamptothecin (9NC) significantly inhibit the uptake activity of OAT3. 9NC also inhibited the substrate transport mediated by OAT1. The substrate uptakes of OAT1, OCTN1 and OCTN2 were significantly decreased in the presence of CZ112, while CPT-11 potently down-regulated the transport activity of OCT1 and OCT3. 5. In summary, our study demonstrated that CPT and its eight derivatives selectively inhibit the substrate uptakes mediated by the essential SLCs. This information contributes to understanding the localized toxicity of CPTs and provides novel molecular targets for the therapeutic optimization of CPTs in the future.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Camptotecina/farmacologia , Transportadores de Ânions Orgânicos/metabolismo , Interações Medicamentosas , Células HEK293 , Humanos
19.
J Exp Bot ; 66(3): 879-88, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25399005

RESUMO

Dihydroxyacid dehydratase (DHAD) catalyses a key step in the branched-chain amino acid (BCAA) biosynthetic pathway that exists in numerous organisms, including bacteria, fungi, and plants, but not humans. In Arabidopsis thaliana, DHAD is encoded by a single gene (AT3G23940), but its biological function in controlling plant development remains uncharacterized. In this study, we showed that DHAD is highly expressed in most vegetative and reproductive tissues. It is an essential gene, and complete disruption caused partial sterility in both male and female gametophyte phases. In addition, reduced expression of DHAD in knockdown mutants resulted in a reduction in the accumulation of all three BCAAs in roots and, as a consequence, led to a shorter root phenotype, which could be restored by an exogenous supplement of free BCAAs. Interestingly, the knockdown mutants became hypersensitive to salt stress, not to heavy metal stress, implying that BCAAs may act as osmolytes in salt tolerance. This would be the second amino acid shown to confer such a function in addition to the well-documented proline. Our results provide evidence that BCAA biosynthesis plays important roles in gametophyte and root development, and BCAA homeostasis contributes to the adaptation of Arabidopsis to salinity stress.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Células Germinativas Vegetais/enzimologia , Hidroliases/genética , Salinidade , Estresse Fisiológico , Arabidopsis/efeitos dos fármacos , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Células Germinativas Vegetais/efeitos dos fármacos , Células Germinativas Vegetais/crescimento & desenvolvimento , Hidroliases/metabolismo , Cloreto de Sódio/farmacologia
20.
Altern Ther Health Med ; 21(1): 36-44, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25599431

RESUMO

CONTEXT: Diabetic nephropathy (DN) is the main cause of end-stage kidney disease in developed countries. Current therapy can slow the rate of progression of DN, but eventually end-stage renal failure will occur in a proportion of patients. Identification of new strategies and additional complementary and alternative therapies for treating DN are important. OBJECTIVE: The research team wanted to assess the beneficial and harmful effects of using puerarin plus angiotensin converting enzyme inhibitor (ACEI) compared with using only ACEI for treatment of individuals with stage III DN. DESIGN: The research team performed a meta-analysis of randomized, controlled trials (RCTs) by searching the following electronic databases: (1) the Cochrane Database of Systematic Reviews, (2) the Cochrane Central Register of Controlled Trials (CENTRAL), (3) PubMed, (4) EMBASE (Elsevier), (5) the Allied and Complementary Medicine Database (AMED), (6) the Chinese Biomedicine Database (CBM), (7) the China National Knowledge Infrastructure (CNKI), and (8) the Chinese Biomedical Journals (VIP), with no language restrictions, as well as databases of clinical trials. OUTCOME MEASURES: Measured outcomes included (1) urinary protein measured as urinary albumin excretion rate (UAER) (µg/min) and 24-h urine protein (24-h UP) (mg/24 h); (2) renal function measured as blood urea nitrogen (BUN) (mmol/L) and serum creatinine (SCr) (µmol/L); (3) α1-microglobulin (α1-MG) (mg/24 h) and endothelin-1 (ET-1) (ng/24 h); (4) end points (EPs); and (5) adverse events (AEs). RESULTS: Ten RCTs involving 669 participants were included. All trials were conducted and published in China. Treatment of DN with puerarin plus ACEI significantly decreased the UAER-P < .0001, MD = -23.43 (95% CI, -33.95 to -12.91), and had no effect on 24-h UP-P = .09, MD = -56.76 (95% CI, -122.65 to 9.12); BUN-P = .17, MD = -0.51 (95% CI, -1.24 to 0.21); and SCr-P = .26, MD = -4.43 (95% CI, -12.05 to 3.20). One trial reported abdominal discomfort and nausea (2 cases) in the treatment group. CONCLUSIONS: Puerarin may be a beneficial therapy for treating DN; however, this hypothesis needs to be proven by additional high-quality studies using large samples and multicenter evidence.


Assuntos
Nefropatias Diabéticas/tratamento farmacológico , Isoflavonas/efeitos adversos , Isoflavonas/uso terapêutico , Vasodilatadores/efeitos adversos , Vasodilatadores/uso terapêutico , Adulto , Idoso , Idoso de 80 Anos ou mais , Albuminúria , Nitrogênio da Ureia Sanguínea , Creatinina/sangue , Nefropatias Diabéticas/epidemiologia , Nefropatias Diabéticas/fisiopatologia , Humanos , Pessoa de Meia-Idade
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