RESUMEN
In this study, we propose a novel method for identifying lithology using an attention mechanism-enhanced graph convolutional neural network (AGCN). The aim of this method is to address the limitations of traditional approaches that evaluate unbalanced lithology by improving the identification of thin layers and small samples, while providing reliable data support for reservoir evaluation. To achieve this goal, we begin by using Principal Component Analysis (PCA) with maximum and minimum distance clustering (Max-min-distance) to correct the logging curves, which compensates for the low resolution of thin layers and enhances the accuracy of stratigraphic representation. Subsequently, we transform the logging data into graph-structured data by connecting distance similarity points and feature similarity points of the logging samples. We then use the graph convolutional network (GCN) to identify lithology, leveraging both labeled and unlabeled data to enhance the ability to identify lithology in small sample datasets. Additionally, our model incorporates a channel and spatial attention mechanism that assigns weights to the graph structure during lithology identification, improving the model's capability to discern differences across samples. To evaluate the performance of our model, we constructed a lithology dataset comprising five wells and conducted experiments. The results indicate that our approach achieves a maximum accuracy of 97.67%, surpassing the performance of a singlestructure model in lithology identification. In conclusion, our proposed method provides a promising and effective approach for unbalanced lithology identification, significantly improving accuracy levels.
RESUMEN
Ferroptosis, a form of regulated cell death that is driven by iron-dependent phospholipid peroxidation, has been implicated in multiple diseases, including cancer1-3, degenerative disorders4 and organ ischaemia-reperfusion injury (IRI)5,6. Here, using genome-wide CRISPR-Cas9 screening, we identified that the enzymes involved in distal cholesterol biosynthesis have pivotal yet opposing roles in regulating ferroptosis through dictating the level of 7-dehydrocholesterol (7-DHC)-an intermediate metabolite of distal cholesterol biosynthesis that is synthesized by sterol C5-desaturase (SC5D) and metabolized by 7-DHC reductase (DHCR7) for cholesterol synthesis. We found that the pathway components, including MSMO1, CYP51A1, EBP and SC5D, function as potential suppressors of ferroptosis, whereas DHCR7 functions as a pro-ferroptotic gene. Mechanistically, 7-DHC dictates ferroptosis surveillance by using the conjugated diene to exert its anti-phospholipid autoxidation function and shields plasma and mitochondria membranes from phospholipid autoxidation. Importantly, blocking the biosynthesis of endogenous 7-DHC by pharmacological targeting of EBP induces ferroptosis and inhibits tumour growth, whereas increasing the 7-DHC level by inhibiting DHCR7 effectively promotes cancer metastasis and attenuates the progression of kidney IRI, supporting a critical function of this axis in vivo. In conclusion, our data reveal a role of 7-DHC as a natural anti-ferroptotic metabolite and suggest that pharmacological manipulation of 7-DHC levels is a promising therapeutic strategy for cancer and IRI.
Asunto(s)
Deshidrocolesteroles , Ferroptosis , Humanos , Membrana Celular/metabolismo , Colesterol/biosíntesis , Colesterol/metabolismo , Sistemas CRISPR-Cas/genética , Deshidrocolesteroles/metabolismo , Genoma Humano , Enfermedades Renales/metabolismo , Membranas Mitocondriales/metabolismo , Metástasis de la Neoplasia , Neoplasias/metabolismo , Neoplasias/patología , Fosfolípidos/metabolismo , Daño por Reperfusión/metabolismoRESUMEN
Mechanistic target of rapamycin complex 1 (mTORC1) is a conserved serine/threonine kinase that integrates various environmental signals to regulate cell growth and metabolism. mTORC1 activation requires tethering to lysosomes by the Ragulator-Rag complex. However, the dynamic regulation of the interaction between Ragulator and Rag guanosine triphosphatase (GTPase) remains unclear. In this study, that LAMTOR1, an essential component of Ragulator, is dynamically ubiquitinated depending on amino acid abundance is reported. It is found that the E3 ligase TRAF4 directly interacts with LAMTOR1 and catalyzes the K63-linked polyubiquitination of LAMTOR1 at K151. Ubiquitination of LAMTOR1 by TRAF4 promoted its binding to Rag GTPases and enhanced mTORC1 activation, K151R knock-in or TRAF4 knock-out blocks amino acid-induced mTORC1 activation and accelerates the development of inflammation-induced colon cancer. This study revealed that TRAF4-mediated LAMTOR1 ubiquitination is a regulatory mechanism for mTORC1 activation and provides a therapeutic target for diseases involving mTORC1 dysregulation.
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Neoplasias Colorrectales , Proteínas de Unión al GTP Monoméricas , Humanos , Diana Mecanicista del Complejo 1 de la Rapamicina/genética , Proteínas de Unión al GTP Monoméricas/genética , Proteínas de Unión al GTP Monoméricas/metabolismo , Factor 4 Asociado a Receptor de TNF/metabolismo , Ubiquitinación , Aminoácidos/metabolismoRESUMEN
Cyclic GMP-AMP synthase (cGAS) is the major sensor for cytosolic DNA and activates type I interferon signaling and plays an essential role in antitumor immunity. However, it remains unclear whether the cGAS-mediated antitumor activity is affected by nutrient status. Here, our study reports that methionine deprivation enhances cGAS activity by blocking its methylation, which is catalyzed by methyltransferase SUV39H1. We further show that methylation enhances the chromatin sequestration of cGAS in a UHRF1-dependent manner. Blocking cGAS methylation enhances cGAS-mediated antitumor immunity and suppresses colorectal tumorigenesis. Clinically, cGAS methylation in human cancers correlates with poor prognosis. Thus, our results indicate that nutrient stress promotes cGAS activation via reversible methylation, and suggest a potential therapeutic strategy for targeting cGAS methylation in cancer treatment.
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Cromatina , Metionina , Humanos , Cromatina/genética , Metionina/genética , Nucleotidiltransferasas/genética , Nucleotidiltransferasas/metabolismo , ADN , Inmunidad Innata , Desmetilación , Proteínas Potenciadoras de Unión a CCAAT/genética , Ubiquitina-Proteína Ligasas/genéticaRESUMEN
Ferroptosis is a lipid peroxidation-dependent cell death caused by metabolic dysfunction. Ferroptosis-associated enzymes are promising therapeutic targets for cancer treatment. However, such therapeutic strategies show limited efficacy due to drug resistance and other largely unknown underlying mechanisms. Here we report that cystine transporter SLC7A11 is upregulated in lung cancer stem-like cells (CSLC) and can be activated by stem cell transcriptional factor SOX2. Mutation of SOX2 binding site in SLC7A11 promoter reduced SLC7A11 expression and increased sensitivity to ferroptosis in cancer cells. Oxidation at Cys265 of SOX2 inhibited its activity and decreased the self-renewal capacity of CSLCs. Moreover, tumors with high SOX2 expression were more resistant to ferroptosis, and SLC7A11 expression was positively correlated with SOX2 in both mouse and human lung cancer tissue. Together, our study provides a mechanism by which cancer cells evade ferroptosis and suggests that oxidation of SOX2 can be a potential therapeutic target for cancer treatment. SIGNIFICANCE: This study uncovers a SOX2-SLC7A11 regulatory axis that confers resistance to ferroptosis in lung cancer stem-like cells.
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Sistema de Transporte de Aminoácidos y+/metabolismo , Biomarcadores de Tumor/metabolismo , Ferroptosis , Regulación Neoplásica de la Expresión Génica , Neoplasias Pulmonares/patología , Células Madre Neoplásicas/patología , Factores de Transcripción SOXB1/metabolismo , Sistema de Transporte de Aminoácidos y+/genética , Animales , Apoptosis , Biomarcadores de Tumor/genética , Proliferación Celular , Humanos , Peroxidación de Lípido , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Ratones , Mutación , Células Madre Neoplásicas/metabolismo , Pronóstico , Factores de Transcripción SOXB1/genética , Tasa de Supervivencia , Activación Transcripcional , Células Tumorales CultivadasRESUMEN
Spaceflight-associated immune system weakening ultimately limits the ability of humans to expand their presence beyond the earth's orbit. A mechanistic study of microgravity-regulated immune cell function is necessary to overcome this challenge. Here, we demonstrate that both spaceflight (real) and simulated microgravity significantly reduce macrophage differentiation, decrease macrophage quantity and functional polarization, and lead to metabolic reprogramming, as demonstrated by changes in gene expression profiles. Moreover, we identified RAS/ERK/NFκB as a major microgravity-regulated pathway. Exogenous ERK and NFκB activators significantly counteracted the effect of microgravity on macrophage differentiation. In addition, microgravity also affects the p53 pathway, which we verified by RT-qPCR and Western blot. Collectively, our data reveal a new mechanism for the effects of microgravity on macrophage development and provide potential molecular targets for the prevention or treatment of macrophage differentiation deficiency in spaceflight.
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Sistema de Señalización de MAP Quinasas , Macrófagos/metabolismo , Redes y Vías Metabólicas , FN-kappa B/metabolismo , Vuelo Espacial , Simulación de Ingravidez , Proteínas ras/metabolismo , Animales , Diferenciación Celular , Polaridad Celular , Regulación hacia Abajo/genética , Perfilación de la Expresión Génica , Células Madre Hematopoyéticas/citología , Células Madre Hematopoyéticas/metabolismo , Masculino , Ratones Endogámicos C57BL , Transducción de Señal , Transcripción Genética , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
Ovarian cancer is the most lethal malignant tumor of female reproductive system. It is well-known that induction of STING-mediated type I interferons can enhance the resultant antitumor activity. However, STING pathway is usually inactivated in cancer cells at multiple levels. Here, we identified deubiquitinase USP35 is upregulated in ovarian cancer tissues. High level of USP35 was correlated with diminished CD8+ T cell infiltration and poor prognosis in ovarian cancer patients. Mechanistically, we found that silencing USP35 reinforces the activation of STING-TBK1-IRF3 pathway and promotes the expression of type I interferons. Our data further showed that USP35 can directly deubiquitinate and inactivate STING. Interestingly, activation of STING promotes its binding to USP35 in a STING phosphorylation-dependent manner. Functionally, we found that knockdown of USP35 sensitizes ovarian cancer cells to the DNA-damage chemotherapeutic drug cisplatin. Overall, our study indicates that upregulation of USP35 may be a mechanism of the restricted STING activity in cancer cells, and highlights the significance of USP35 as a potential therapeutic target for ovarian cancer.
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Endopeptidasas/metabolismo , Interferón Tipo I/metabolismo , Neoplasias Ováricas/genética , Transducción de Señal/efectos de los fármacos , Animales , Antineoplásicos/uso terapéutico , Cisplatino/uso terapéutico , Daño del ADN/efectos de los fármacos , Endopeptidasas/genética , Femenino , Humanos , Factor 3 Regulador del Interferón/metabolismo , Proteínas de la Membrana/metabolismo , Ratones , Ratones Endogámicos C57BL , Neoplasias Ováricas/tratamiento farmacológico , Neoplasias Ováricas/metabolismo , Neoplasias Ováricas/patología , Fosforilación , Proteínas Serina-Treonina Quinasas/metabolismo , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Polygala tenuifolia Willd. is a traditional Chinese herbal medicine that is widely used in treating nervous system disorders. Triterpene saponins in P. tenuifolia (polygala saponins) have excellent biological activity. As a precursor for the synthesis of presenegin, oleanolic acid (OA) plays an important role in the biosynthesis of polygala saponins. However, the mechanism behind the biosynthesis of polygala saponins remains to be elucidated. In this study, we found that CYP716A249 (GenBank: ASB17946) oxidized the C-28 position of ß-amyrin to produce OA. Using quantitative real-time PCR, we observed that CYP716A249 had the highest expression in the roots of 2-year-old P. tenuifolia, which provided a basis for the selection of samples for gene cloning. To identify the function of CYP716A249, the strain R-BE-20 was constructed by expressing ß-amyrin synthase in yeast. Then, CYP716A249 was co-expressed with ß-amyrin synthase to construct the strain R-BPE-20 by using the lithium acetate method. Finally, we detected ß-amyrin and OA by ultra-HPLC-Q Exactive hybrid quadrupole-Orbitrap high-resolution accurate mass spectrometry and GC-MS. The results of this study provide insights into the biosynthesis pathway of polygala saponins.
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Clonación Molecular/métodos , Polygala/genética , Polygala/metabolismo , Proteínas de Saccharomyces cerevisiae/biosíntesis , Proteínas de Saccharomyces cerevisiae/genética , Triterpenos/metabolismo , Proteínas de Arabidopsis/biosíntesis , Proteínas de Arabidopsis/genética , Sistema Enzimático del Citocromo P-450/biosíntesis , Sistema Enzimático del Citocromo P-450/genética , Regulación de la Expresión Génica de las Plantas , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/genética , Ácido Oleanólico/metabolismo , Filogenia , Saccharomyces cerevisiae , Saponinas/biosíntesis , Saponinas/genéticaAsunto(s)
Aminoaciltransferasas/metabolismo , Antígenos de Diferenciación/metabolismo , Antígeno CD47/metabolismo , Receptores Inmunológicos/metabolismo , Aminoaciltransferasas/análisis , Aminoaciltransferasas/deficiencia , Antígenos de Diferenciación/química , Antígeno CD47/química , Antígeno CD47/deficiencia , Humanos , Isoenzimas/análisis , Isoenzimas/deficiencia , Isoenzimas/metabolismo , Receptores Inmunológicos/químicaRESUMEN
Exposure to spaceflight and microgravity causes physiologic and psychologic changes including bone loss, cardiovascular dysfunction, and immune dysfunction. Anemia and hematopoietic disorders are observed in astronauts after spaceflight. Hematopoietic stem and progenitor cells (HSPCs), which can self-renew and give rise to all blood cells, play vital roles in hematopoiesis and homeostasis; however, the molecular mechanisms responsible for the impacts of microgravity on the proliferation of HSPCs remain unclear. We maintained mouse bone marrow HSPCs in the presence of stem cell factor for 12 d under spaceflight and simulated microgravity conditions, respectively, and analyzed cell proliferation and gene expression. Both spaceflight and simulated microgravity significantly decreased the number of HSPCs, mainly by blocking cell cycle at G1/S transition, but did not affect their differentiation abilities. RNA-sequencing data indicated that genes related to cell proliferation were down-regulated, whereas the genes related to cell death were up-regulated under microgravity. Among the gene signatures, we identified that the Kit-Ras/cAMP-cAMP response element-binding protein pathway might be one of the major microgravity-regulated pathways during HSPC proliferation. Furthermore, the quantification of notable genes was validated at the mRNA levels under simulated microgravity condition. Overall, these results would help us to understand the intracellular molecular mechanisms regulating microgravity-inhibited proliferation of HSPCs.-Wang, P., Tian, H., Zhang, J., Qian, J., Li, L., Shi, L., Zhao, Y. Spaceflight/microgravity inhibits the proliferation of hematopoietic stem cells by decreasing Kit-Ras/cAMP-CREB pathway networks as evidenced by RNA-Seq assays.
Asunto(s)
Células de la Médula Ósea/citología , Células Madre Hematopoyéticas/citología , Vuelo Espacial , Simulación de Ingravidez , Ingravidez , Animales , Diferenciación Celular/genética , Proliferación Celular , Células Cultivadas , AMP Cíclico/metabolismo , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Hematopoyesis , Ratones , Ratones Endogámicos C57BL , Proteínas Proto-Oncogénicas c-kit/metabolismo , RNA-Seq , Transducción de Señal , Proteínas ras/metabolismoRESUMEN
Discrimination of species and geographical origins of traditional Chinese medicine (TCM) is essential to prevent adulteration and inferior problems. We studied Ephedra sinica Stapf, Ephedra intermedia Schrenk et C.A.Mey. and Ephedra przewalskii Bge. to investigate the relationship between inorganic element content and these three species and their geographical origins. 38 elemental fingerprints from six major Ephedra-producing regions, namely, Inner Mongolia, Ningxia, Gansu, Shanxi, Shaanxi, and Sinkiang, were determined to evaluate the importance of inorganic elements to three species and their geographical origins. The contents of 15 elements, namely, N, P, K, S, Ca, Mg, Fe, Mn, Na, Cl, Sr, Cu, Zn, B, and Mo, of Ephedra samples were measured using inductively coupled plasma mass spectroscopy. Elemental contents were used as chemical indicators to classify species and origins of Ephedra samples using a radar plot and multivariate data analysis, including hierarchical cluster analysis (HCA), principal component analysis (PCA), and discriminant analysis (DA). Ephedra samples from different species and geographical origins could be differentiated. This study showed that inorganic elemental fingerprint combined with multivariate statistical analysis is a promising tool for distinguishing three Ephedra species and their geographical origins, and this strategy might be an effective method for authenticity discrimination of TCM.
Asunto(s)
Compuestos Inorgánicos de Carbono/análisis , Compuestos Inorgánicos de Carbono/metabolismo , Ephedra/clasificación , Ephedra/metabolismo , Espectrometría de Masas/métodos , Análisis Discriminante , Geografía , Análisis de Componente PrincipalRESUMEN
Macrophages acquire distinct phenotypes during tissue stress and inflammatory responses. Macrophages are roughly categorized into two different subsets named inflammatory M1 and anti-inflammatory M2 macrophages. We herein identified a unique pathogenic macrophage subpopulation driven by IL-23 with a distinct gene expression profile including defined types of cytokines. The freshly isolated resting mouse peritoneal macrophages were stimulated with different cytokines in vitro, the expression of cytokines and chemokines were detected by microarray, real-time PCR, ELISA and multiple colors flow cytometry. Adoptive transfer of macrophages and imiquimod-induced psoriasis mice were used. In contrast to M1- and M2-polarized macrophages, IL-23-treated macrophages produce large amounts of IL-17A, IL-22 and IFN-γ. Biochemical and molecular studies showed that IL-23 induces IL-17A expression in macrophages through the signal transducer and activator of transcription 3 (STAT3)-retinoid related orphan receptor-γ T (RORγT) pathway. T-bet mediates the IFN-γ production in IL-23-treated macrophages. Importantly, IL-23-treated macrophages significantly promote the dermatitis pathogenesis in a psoriasis-like mouse model. IL-23-treated resting macrophages express a distinctive gene expression prolife compared with M1 and M2 macrophages. The identification of IL-23-induced macrophage polarization may help us to understand the contribution of macrophage subpopulation in Th17-cytokines-related pathogenesis.
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Polaridad Celular , Imiquimod , Interleucina-23/metabolismo , Macrófagos/metabolismo , Psoriasis/inducido químicamente , Psoriasis/patología , Animales , Macrófagos/patología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Psoriasis/metabolismoRESUMEN
The quality control of Polygala tenuifolia Wild. is a major challenge in its clinical application. In this paper, a new strategy for the quality evaluation of P. tenuifolia extracts was verified through reverse-phase ultra-performance liquid chromatography (UPLC). The quantitative analysis of multi-components by a single marker (QAMS) was conducted with 3,6'-disinapoyl sucrose as an internal reference substance. Eight components (i.e., sibiricose A5, sibiricose A6, glomeratose A, tenuifoliside A, tenuifoliside B, tenuifoliside C, sibiricaxanthone B, and polygalaxanthone III) were determined based on the relative correction factors. The concentrations of these components were also determined by applying a conventional external standard method. The cosine value confirmed the consistency of the two methods (cosine ratio value >0.999920). Hierarchical cluster analysis, radar plots, and discriminant analysis were performed to classify 23 batches of P. tenuifolia extracts from Shanxi, Hebei, and Shaanxi in China. Results revealed that QAMS combined with radar plots and multivariate data analysis could accurately measure and clearly distinguish the different quality samples of P. tenuifolia. Hence, QAMS is a feasible and promising method for the quality control of P. tenuifolia.
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Carbohidratos/análisis , Cromatografía Líquida de Alta Presión/métodos , Cromatografía de Fase Inversa/métodos , Extractos Vegetales/análisis , Polygala/química , Carbohidratos/química , Carbohidratos/aislamiento & purificación , Estructura Molecular , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Raíces de Plantas/química , Control de CalidadRESUMEN
This work was launched to explore the effect of habitat and growth year on the secondary metabolites contents of cultivated Polygala tenuifolia. The samples of cultivated P. tenuifolia were analyzed by ultra-high performance liquid chromatography(UPLC)-quadrupole time-of-flight mass spectrometry(Q-TOF MS), and the obtained data were analyzed using multiple statistical analysis and cluster analysis. The results showed that compared with growth year, habitat is a main influencing factor which affected the secondary metabolites contents of P. tenuifolia. The contents of sucrose esters and oligosacchride multi-esters are greatly dependent on the habitat (the sample-AG with high levels of components of tenuifoliside B and tenuifoliside C, and the sample-FY with high levels of 3,6'-disinapoyl sucrose, tenuifoliose S, tenuifoliose L, and tenuifoliose V). There is no obvious effect of habitat and growth year on xanthone. The contents of triterpene saponins are greatly dependent on the growth year, and the content of parts of triterpene saponins increased as time goes on.The result indicated that the effect of habitat and growth year on different types of secondary metabolites is not completely equivalent. This study will contribute to the breeding of P. tenuifolia and amendment of current commodity criteria.
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Polygala/química , Saponinas/análisis , Triterpenos/análisis , Cromatografía Líquida de Alta Presión , Ecosistema , Ésteres/análisis , Espectrometría de Masas , Oligosacáridos/análisis , Fitoquímicos/análisis , Metabolismo SecundarioRESUMEN
Inorganic elements are important components of medicinal herbs, and provide valuable experimental evidence for the quality evaluation and control of traditional Chinese medicine (TCM). In this study, to investigate the relationship between the inorganic elemental fingerprint and geographical origin identification of cultivated Polygala tenuifolia, 41 elemental fingerprints of P. tenuifolia from four major polygala-producing regions (Shanxi, Hebei, Henan, and Shaanxi) were evaluated to determine the importance of inorganic elements to cultivated P. tenuifolia. A total of 15 elemental (B, Ca, Cl, Cu, Fe, K, Mg, Mn, Na, N, Mo, S, Sr, P, and Zn) concentrations of cultivated P. tenuifolia were measured using inductively coupled plasma mass spectroscopy (ICP-MS). The element composition samples were classified by radar plot, elemental fingerprint, and multivariate data analyses, such as hierarchical cluster analysis (HCA), principle component analysis (PCA), and discriminant analysis (DA). This study shows that radar plots and multivariate data analysis can satisfactorily distinguish the geographical origin of cultivated P. tenuifolia. Furthermore, PCA results revealed that N, Cu, K, Mo, Sr, Ca, and Zn are the characteristic elements of cultivated P. tenuifolia. Therefore, multi-element fingerprinting coupled with multivariate statistical techniques can be considered an effective tool to discriminate geographical origin of cultivated P. tenuifolia.
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Plantas Medicinales/química , Polygala/química , Oligoelementos/química , Análisis Discriminante , Geografía , Espectrometría de Masas , Plantas Medicinales/clasificación , Polygala/clasificación , Análisis de Componente Principal , Análisis Espectral , Oligoelementos/aislamiento & purificaciónRESUMEN
As one of the most important traditional Chinese medicine, the quality of Polygala tenuifolia is difficult to control and a new method must be established to facilitate/assist the breeding of P. tenuifolia. In this study, UPLC/Q-TOF-MS-based metabolomics analysis was performed to determine the chemical composition and screen metabolite biomarkers according to agronomic traits. A total of 29 compounds and 18 metabolite biomarkers were found. AFLP-based marker-assisted selection (MAS) was used to identify molecular marker bands and screen characteristic bands associated with specific agronomic traits. 184 bands and 76 characteristic AFLP bands were found. The correlation network between compounds and characteristic AFLP bands was built, so we may directly breed certain P. tenuifolia herbs with special agronomic traits (or characteristic AFLP bands), which exhibit specific pharmacological functions depending on the content of the active compounds. The proposed method of metabolomics coupled with MAS could facilitate/assist the breeding of P. tenuifolia.
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Análisis del Polimorfismo de Longitud de Fragmentos Amplificados , Metaboloma , Fitomejoramiento , Polygala/crecimiento & desarrollo , Cromatografía Líquida de Alta Presión , Análisis por Conglomerados , ADN de Plantas/genética , Espectrometría de Masas , Metabolómica , Raíces de Plantas/genética , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/metabolismo , Polygala/genética , Polygala/metabolismoRESUMEN
The agronomic traits (plant height, root diameter, root length, first lateral root height, lateral root amount, root weight) of 18 Polygala tenuifolia samples with different agronomic traits were analyzed, respectively. HPLC was used to analyze three main characteristic components including tenuifolin, polygalaxanthone â ¢, and 3,6'-disinapoyl sucrose. At last, the correlation between six agronomic traits and three main characteristic components were analyzed by scatter plot. We found no significant correlation between root diameter and three main characteristic components. There were no obvious correlations between tenuifolin and the remaining five agronomic traits. Short root length and first lateral root height as well as high lateral root amount resulted in high levels of polygalaxanthone â ¢ in P. tenuifolia samples. High levels of 3,6'-disinapoyl sucrose were observed in P. tenuifolia samples with longer root. So, the current commodity criteria and traditional breeding of P. tenuifolia did not conform to pharmacopoeia standards, which excellent medicinal materials should have high contents of the main characteristic components. It was urgent to revise the current commodity criteria and breeding methods.
Asunto(s)
Fitomejoramiento , Polygala , Cromatografía Líquida de Alta Presión , Ácidos Cumáricos/análisis , Diterpenos de Tipo Kaurano/análisis , Medicamentos Herbarios Chinos/química , Glicósidos/análisis , Raíces de Plantas/crecimiento & desarrollo , Plantas Medicinales/química , Plantas Medicinales/crecimiento & desarrollo , Polygala/química , Polygala/crecimiento & desarrollo , Sacarosa/análogos & derivados , Sacarosa/análisis , Xantonas/análisisRESUMEN
The content changes of chemical components in different phenological phase of the cultivated Polygala tenuifolia is one of the important factors for determination of the best harvest time in the production practice. In this study, the digital gene expression (DGE) profiles of the cultivated P. tenuifolia were analyzed in different phenological phase (flowering fruit bearing stage, wilting stage, dormancy stage). The differentially expressed genes were found in the biosynthesis of chemical composition in P. tenuifolia, and the representational ones were validated by RT-q PCR. Then, the key enzymes(CYP450s and UGTs) involved in the downstream of the triterpenoid saponins biosynthesis pathway in P. tenuifolia were predicted through the correlation analysis of gene expression. The number of down-regulated genes was more than that of up-regulated in P. tenuifolia from flowering fruit bearing stage to dormancy stage. Six differentially expressed genes (HMGS, PMK, FPPS, SQS, SE, ß-AS) and five (PAL, C4 H, 4CL, CAD, peroxidase) were annotated to the triterpenoid saponins and phenylpropanoid biosynthesis pathway in P. tenuifolia, respectively. Compared to wilting and dormancy stages, the saponins, xanthones, and lignins were largely synthesized at the flowering fruit bearing stage of P. tenuifolia. Furthermore, UGT83A1, CYP716B1, CYP98A3, CYP86B1, and CYP94A1 may be the part of key enzymes in the downstream of the triterpenoid saponins biosynthesis pathway in P. tenuifolia. This study provides evidence to support the correctness of traditional harvest time of P. tenuifolia at the level of transcription, and lays the scientific foundation for gene cloning and functional verification of CYP450 s and UGTs in the downstream of the triterpenoid saponins biosynthesis pathway in P. tenuifolia in the future.
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Polygala/genética , Transcriptoma , Sistema Enzimático del Citocromo P-450/metabolismo , Flores , Frutas , Glucuronosiltransferasa/metabolismo , Lignina/biosíntesis , Latencia en las Plantas , Saponinas/biosíntesis , Triterpenos/metabolismo , Xantonas/metabolismoRESUMEN
Radix polygalae, the dried roots of Polygala tenuifolia and P. sibirica, is one of the most well-known traditional Chinese medicinal plants. Radix polygalae contains various saponins, xanthones, and oligosaccharide esters and these compounds are responsible for several pharmacological properties. To provide basic breeding information, enhance molecular biological analysis, and determine secondary metabolite biosynthetic pathways of P. tenuifolia, we applied Illumina sequencing technology and de novo assembly. We also applied this technique to gain an overview of P. tenuifolia transcriptome from samples with different years. Using Illumina sequencing, approximately 67.2% of unique sequences were annotated by basic local alignment search tool similarity searches against public sequence databases. We classified the annotated unigenes by using Nr, Nt, GO, COG, and KEGG databases compared with NCBI. We also obtained many candidates CYP450s and UGTs by the analysis of genes in the secondary metabolite biosynthetic pathways, including putative terpenoid backbone and phenylpropanoid biosynthesis pathway. With this transcriptome sequencing, future genetic and genomics studies related to the molecular mechanisms associated with the chemical composition of P. tenuifolia may be improved. Genes involved in the enrichment of secondary metabolite biosynthesis-related pathways could enhance the potential applications of P. tenuifolia in pharmaceutical industries.
RESUMEN
OBJECTIVE AND DESIGN: Molecular mechanisms of microgravity-caused immunosuppression are not fully elucidated. In the present study, we investigated the effects of simulated microgravity on macrophage functions and tried to identify the related intracellular signal pathways. MATERIAL OR SUBJECTS: Primary mouse macrophages were used in the present study. The gene expression and function of IL-4-treated mouse macrophages were detected after simulated microgravity or 1 g control. METHODS: Freshly isolated primary mouse macrophages were cultured in a standard simulated microgravity situation using a rotary cell culture system (RCCS-1) and 1 g control conditions. Real-time PCR, western blots and flow cytometry were used to investigate the related intracellular signals and molecule expression. RESULTS: The arginase mRNA and protein levels in freshly isolated primary mouse macrophages under simulated microgravity using RCCS-1 were significantly higher than those under normal gravity. Meanwhile, simulated microgravity induced over-expression of C/EBPß, a transcription factor of arginase promoter, and activation of p38 MAPK, which could increase C/EBPß expression. Furthermore, up-regulation of Interleukin-6 (IL-6) and down-regulation of IL-12 p40 (IL-12B) in LPS-stimulated macrophages were also detected after simulated microgravity, which is regulated by C/EBPß. CONCLUSIONS: Simulated microgravity activates a p38 MAPK-C/EBPß pathway in macrophages to up-regulate arginase and IL-6 expression and down-regulate IL-12B expression. Both increased arginase expression and decreased IL-12B expression in macrophages during inflammation could result in immunosuppression under microgravity.