RESUMEN
Functional nanomaterials such as iron oxide nanoparticles have been extensively explored for the diagnosis and treatment of central nervous system diseases. However, an insufficient understanding of the comprehensive nanomaterial-biological interactions in the brain hinders the nanomaterials from meeting the medical requirements for translational research. Here, FDA-approved ferumoxytol, an iron oxide nanoparticle, is chosen as the model nanomaterial for a systematic study of the dynamic interactions between ferumoxytol and immune cells, including microglia and macrophages, in the brain tumors. Strikingly, up to 90% of intratumorally injected ferumoxytol nanoparticles are recognized and phagocytized by tumor-associated microglia and macrophages. The dynamic trafficking progress of ferumoxytol in microglia and macrophages, including scavenger receptor-mediated endocytosis, lysosomal internalization, and extracellular vesicle-dominated excretion, is further studied. Importantly, the results demonstrate that extracellular vesicle-encapsulated nanoparticles could be gradually eliminated from the brain along with cerebrospinal fluid circulation over 21 days. Moreover, ferumoxytol exhibits no obvious long-term neurological toxicity after its injection. The study suggests that the dynamic biointeractions of nanoparticles with immune cells in the brain exert a key rate-limiting impact on the efficiency of targeting tumor cells and their in vivo fate and thus provide a deeper understanding of the nanomaterials in the brain for clinical applications.
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Neoplasias Encefálicas , Nanopartículas , Encéfalo , Óxido Ferrosoférrico , Humanos , Macrófagos , Imagen por Resonancia MagnéticaRESUMEN
The diagnosis of drowning is one of the most difficult in forensic medicine. Forensic diatomology has been proposed to be useful in solving the diagnosis of drowning and considered to be a reliable indicator of the site of drowning. The Yangtze River and Jialing River are the main rivers in the Chongqing area (China), and a large number of corpses are found in the rivers every year. However, the distribution of diatoms in the rivers was not fully studied. In the presented study, a Microwave Digestion-Vacuum Filtration-Scanning Electron Microscopy (MD-VF-SEM) method was performed to acquire the qualitative and quantitative data of diatoms of water samples collected from 10 different sites of the Yangtze River and Jialing River in Chongqing section during different seasons. Our study not only created the diatomological maps of water bodies in Chongqing section of the Yangtze River and Jialing River for the first time but also identified some seasonal and site-specific diatoms that can be taken as markers of particular sites or seasons of drowning. The results of our study may provide forensic scientists helpful reference in solving the drowning cases.
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Diatomeas/clasificación , Diatomeas/aislamiento & purificación , Ahogamiento/diagnóstico , Medicina Legal/métodos , Ríos/microbiología , Microbiología del Agua , China , Ahogamiento/microbiología , Humanos , Microscopía Electrónica de Rastreo , Imágenes de Microonda , Análisis Multivariante , Estaciones del AñoRESUMEN
Cantharidin is the major bioactive compound extracted from the blister beetle, a traditional Chinese medicine, and has been proved to be a natural component with widely antitumor activity. However, clinical application of cantharidin is relatively restricted due to its potential toxic effects, especially hepatotoxicity. Although cantharidin-induced liver injury has been reported, the underlying molecular mechanisms remain unclear. In the present study, an UPLC-Q-TOF/MS based metabolomics approach combined with blood biochemical analysis, histopathological examination, and cell apoptosis assay were used to investigate the mechanisms of cantharidin-induced hepatotoxicity. A total of 54 significantly changed metabolites and 14 disturbed metabolic pathways were identified in the cantharidin exposed groups. Among them, four metabolites (oxidized glutathione, glutathione, 3-sulfinoalanine, and deoxycholic acid 3-glucuronide) were selected based on their high impact value and potential biological function in the process of liver injury post cantharidin treatment. Our study provides a deeper understanding of the mechanisms of cantharidin-induced hepatotoxicity and may contribute to reduce the liver injury and gain more effective and safe clinical use of cantharidin. In addition, our results also demonstrated that cantharidin could impair multiple biological processes in liver, and future studies will be necessary to reveal the detailed molecular mechanisms of cantharidin-induced hepatotoxicity.
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Antineoplásicos/toxicidad , Cantaridina/toxicidad , Enfermedad Hepática Inducida por Sustancias y Drogas/metabolismo , Animales , Apoptosis/efectos de los fármacos , Línea Celular , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Cromatografía Líquida de Alta Presión , Humanos , Hígado/efectos de los fármacos , Hígado/metabolismo , Hígado/patología , Masculino , Espectrometría de Masas , Metabolómica/métodos , RatonesRESUMEN
Mycobacterium tuberculosis (Mtb) is an intracellular pathogen that triggers several survival mechanisms against the host immune system. Many studies show that the diverse components of Mtb can modulate apoptosis in various types of cells differently. So far, apoptosis induced by ESAT-6, an early secreted antigenic target of 6-kDa of Mtb, has been studied but the details of molecular mechanism and signaling pathway remain incompletely defined. This study investigated the role of recombinant ESAT-6 in inducing apoptosis in primary bone marrow-derived macrophages (BMDMs) of mice using Annexin V/PI assay with FACS analysis and Western blotting technique. It has been found that ESAT-6-induced apoptosis in BMDMs in a dose- and time-dependent pattern. Apoptosis induced by ESAT-6 was mainly via the intrinsic pathway with elevated protein levels of cleaved caspase-9 and -3. Furthermore, ESAT-6 also induced Bim activation during this process. Interestingly, this event was TLR2-dependent since the effect of ESAT-6 on apoptosis vanished in BMDM from mice with TLR2 deficiency. Furthermore, ROS generation and MAPKs phosphorylation induced by ESAT-6 were also involved in caspase-9 and caspase-3 activation. Taken together, these data suggest that ESAT-6-mediated apoptosis is involved in ROS-MAPKs signaling and further activating the intrinsic pathway, which provides new insights into the basic physiology of macrophage death in tuberculosis.
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Antígenos Bacterianos/farmacología , Apoptosis/efectos de los fármacos , Proteínas Bacterianas/farmacología , Caspasa 3/metabolismo , Caspasa 9/metabolismo , Macrófagos/metabolismo , Mycobacterium tuberculosis/química , Animales , Antígenos Bacterianos/química , Proteínas Bacterianas/química , Células de la Médula Ósea/metabolismo , Células de la Médula Ósea/patología , Macrófagos/patología , Masculino , RatonesRESUMEN
T cell activation is a well-established model for studying cellular responses to exogenous stimulation. Using strand-specific RNA-seq, we observed that intron retention is prevalent in polyadenylated transcripts in resting CD4(+) T cells and is significantly reduced upon T cell activation. Several lines of evidence suggest that intron-retained transcripts are less stable than fully spliced transcripts. Strikingly, the decrease in intron retention (IR) levels correlate with the increase in steady-state mRNA levels. Further, the majority of the genes upregulated in activated T cells are accompanied by a significant reduction in IR. Of these 1583 genes, 185 genes are predominantly regulated at the IR level, and highly enriched in the proteasome pathway, which is essential for proper T cell proliferation and cytokine release. These observations were corroborated in both human and mouse CD4(+) T cells. Our study revealed a novel post-transcriptional regulatory mechanism that may potentially contribute to coordinated and/or quick cellular responses to extracellular stimuli such as an acute infection.
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Linfocitos T CD4-Positivos/metabolismo , Regulación de la Expresión Génica , Intrones/genética , Activación de Linfocitos/genética , Animales , Secuencia Conservada/genética , Humanos , Péptidos y Proteínas de Señalización Intracelular , Ratones , Modelos Biológicos , Complejo de la Endopetidasa Proteasomal/genética , Complejo de la Endopetidasa Proteasomal/metabolismo , Estabilidad del ARN/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Regulación hacia Arriba/genéticaRESUMEN
OBJECTIVE: IL-1ß is a master switch of inflammation and plays an important role in the pathogenesis of vascular disease. During early atherosclerosis development, it is not clearly understood how oxidized low density lipoprotein (oxLDL)induced signaling pathways control NLRP3 inflammasome activation and produce IL-1ß and promote foam cells formation. METHODS: The study used THP-1 macrophage as cell model. Western blot quantified the oxLDL-induced NLRP3 inflammasome related proteins. The FACS detected the expression of SR-A and CD36 receptors on the cells, and caspase-1 activation in the cells. The DCFH-DA assayed the reactive oxygen species (ROS). Oil red O staining techniques examined the intracellular lipid droplet. RESULTS: The OxLDL remarkably increased not only IL-1ß mRNA transcription and pro-IL-1ß protein synthesis but also IL-1ß secretion in human macrophages. The activation of the NLRP3 inflammasome depended on oxLDL-induced generation of ROS, potassium efflux and cathepsin B activity. The OxLDL-induced ROS production that mediates IL-1ß maturation mainly depended on the scavenger receptor of CD36 but not SR-A. The secreted IL-1ß served as an autocrine function for promoting macrophage foam cells formation. CONCLUSIONS: These findings suggest that oxLDL-induced NLRP3 inflammasome activation mainly depends on CD36 involved in the progression of atherosclerosis by promoting oxLDL-mediated inflammation and foam cell formation.
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Antígenos CD36/genética , Proteínas Portadoras/genética , Células Espumosas/citología , Inflamasomas/genética , Lipoproteínas LDL/farmacología , Caspasa 1/metabolismo , Línea Celular Tumoral , Células Cultivadas , Humanos , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Leucocitos Mononucleares/citología , Macrófagos/citología , Proteína con Dominio Pirina 3 de la Familia NLR , ARN Mensajero/metabolismo , ARN Interferente Pequeño/genética , Especies Reactivas de Oxígeno/metabolismo , Receptores Depuradores de Clase A/genéticaRESUMEN
BACKGROUND: Innate immune response constitutes the first line of defense against pathogens. Inflammatory responses involve close contact between different populations of cells. These adhesive interactions mediate migration of cells to sites of infection leading the effective action of cells within the lesions. Cell adhesion molecules are critical to controlling immune response mediating cell adhesion or chemotaxis, as well as coordinating actin-based cell motility during phagocytosis and chemotaxis. Recently, a newly discovered neuroplastin (Np) adhesion molecule is found to play an important role in the nervous system. However, there is limited information on Np functions in immune response. To understand how Np is involved in innate immune response, a mouse model of intraperitoneal infection was established to investigate the effect of Np on macrophage-mediated clearance of E. coli infection and its possible molecular mechanisms. METHODS: Specific deficiency mice with Nptn gene controlling Np65 isoform were employed in this study. The expression levels of mRNA and proteins were detected by qPCR and western blot, or evaluated by flow cytometry. The expression level of NO and ROS were measured with their specific indicators. Cell cycle and apoptosis were detected by specific detection kits. Acid phosphatase activity was measured by flow cytometry after labelling with LysoRed fluorescent probe. Bone marrow derived macrophages (BMDMs) were isolated from bone marrow of mice hind legs. Cell proliferation was detected by CCK8 assay. Cell migration was measured by wound healing assay or transwell assay. RESULTS: The lethal dose of E. coli infection in Np65-/- mice dropped to the half of lethal dose in WT mice. The bacterial load in the spleen, kidney and liver from Np65-/- mice were significantly higher than that from WT mice, which were due to the dramatic reduction of NO and ROS production in phagocytes from Np65-/- mice. Np65 gene deficiency remarkably impaired phagocytosis and function of lysosome in macrophage. Furthermore, Np65 molecule was involved in maturation and proliferation, even in migration and chemotaxis of BMDM in vitro. CONCLUSION: This study for the first time demonstrates that Np is involved in multi-function of phagocytes during bacterial infection, proposing that Np adhesion molecule plays a critical role in clearing pathogen infection in innate immunity.
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Infecciones por Escherichia coli , Escherichia coli , Fosfatasa Ácida , Actinas , Animales , Moléculas de Adhesión Celular , Escherichia coli/metabolismo , Colorantes Fluorescentes , Macrófagos , Glicoproteínas de Membrana/metabolismo , Ratones , Isoformas de Proteínas , ARN Mensajero , Especies Reactivas de OxígenoRESUMEN
Here, we investigated the therapeutic potential of IL-10 by testing its effects on oxLDL-induced lipoprotein uptake and apoptosis by flow cytometry in THP-1-derived macrophages. The mRNA and protein expressions of lipid scavenger receptors (SR-A, CD36) and apoptosis-related proteins (Bcl-2, Bak-1) were also detected. Co-incubation of oxLDL with IL-10 reduced DiI-oxLDL uptake by 16.1±3.8%, 35.2±3.8% and 28.9±1.8% at 6, 12 and 24h of treatment, respectively. Furthermore, treatment with oxLDL for 24h enhanced the SR-A mRNA and protein expressions by 89.3±17.1% and 70.1±17.6%, respectively. IL-10 abrogated the oxLDL-induced SR-A mRNA expression by 50.2±3.9% and its protein by 45.6±1.9%. Meanwhile IL-10 had no effect on the oxLDL-induced increase of CD36 expression. IL-10 inhibited the oxLDL-induced cell apoptosis in a time-dependent manner by 17.3±3.3%, 36.4±2.8% and 31.0±4.3% at 6, 12 and 24h, respectively. OxLDL increased Bak-1 mRNA and protein expressions by 38.4±13.3% and 36.9±12.1%, respectively. However co-stimulation of oxLDL with IL-10 for 24h inhibited Bak-1 expression to 28.4±7.2% (mRNA) and 25.7±6.3% (protein). Meanwhile, IL-10 had no effect on the oxLDL-induced decrease of Bcl-2 expression. Our findings suggested that IL-10 reduced the oxLDL-induced lipoprotein uptake and apoptosis partly via down-regulating the oxLDL-induced expression of SR-A and Bak-1 in THP-1-derived macrophages.
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Regulación hacia Abajo , Interleucina-10/metabolismo , Lipoproteínas LDL/metabolismo , Macrófagos/metabolismo , Receptores Depuradores de Clase A/genética , Proteína Destructora del Antagonista Homólogo bcl-2/genética , Apoptosis , Antígenos CD36/genética , Línea Celular , Humanos , Macrófagos/citología , ARN Mensajero/genéticaRESUMEN
BACKGROUND: Atherosclerosis (AS) is a common cardiovascular disease. Transformation of macrophages to form foam cells by internalizing modified low density-lipoprotein (LDL) via scavenger receptor (SR) is a key pathogenic process in the onset of AS. It has been demonstrated that SR-PSOX functions as either a scavenger receptor for uptake of atherogenic lipoproteins and bacteria or a membrane-anchored chemokine for adhesion of macrophages and T-cells to the endothelium. Therefore, SR-PSOX plays an important role in the development of AS. In this study the key basic amino acids in the chemokine domain of SR-PSOX have been identified for its functions. RESULTS: A cell model to study the functions of SR-PSOX was successfully established. Based on the cell model, a series of mutants of human SR-PSOX were constructed by replacing the single basic amino acid residue in the non-conservative region of the chemokine domain (arginine 62, arginine 78, histidine 80, arginine 82, histidine 85, lysine 105, lysine 119, histidine 123) with alanine (designated as R62A, R78A, H80A, R82A, H85A, K105A, K119A and H123A, respectively). Functional studies showed that the mutants with H80A, H85A, and K105A significantly increased the activities of oxLDL uptake and bacterial phagocytosis compared with the wild-type SR-PSOX. In addition, we have also found that mutagenesis of either of those amino acids strongly reduced the adhesive activity of SR-PSOX by using a highly non-overlapping set of basic amino acid residues. CONCLUSION: Our study demonstrates that basic amino acid residues in the non-conservative region of the chemokine domain of SR-PSOX are critical for its functions. Mutation of H80, H85, and K105 is responsible for increasing SR-PSOX binding with oxLDL and bacteria. All the basic amino acids in this region are important in the cells adhesion via SR-PSOX. These findings suggest that mutagenesis of the basic amino acids in the chemokine domain of SR-PSOX may contribute to atherogenesis.
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Aminoácidos Básicos/genética , Quimiocinas CXC/genética , Quimiocinas CXC/metabolismo , Mutación Puntual/genética , Receptores Depuradores/genética , Receptores Depuradores/metabolismo , Sustitución de Aminoácidos/genética , Carbocianinas/metabolismo , Adhesión Celular , Línea Celular , Membrana Celular/metabolismo , Quimiocina CXCL16 , Escherichia coli/metabolismo , Vectores Genéticos/genética , Proteínas Fluorescentes Verdes/metabolismo , Humanos , Lipoproteínas LDL , Proteínas Mutantes/metabolismo , Fagocitosis , TransfecciónRESUMEN
BACKGROUND: Tuberculosis (TB) is a chronic infectious disease caused by Mycobacterium tuberculosis (Mtb). Granuloma is a pathological feature of tuberculosis and is a tight immune cell aggregation caused by Mtb. The main constituent cells are macrophages and their derivative cells including epithelioid macrophages. However, the molecular mechanism of the transition has not been reported. The purpose of this study was to investigate whether early secreted antigenic target of 6-kDa (ESAT6) can induce the transition of bone marrow-derived macrophages (BMDMs) into epithelioid macrophages and its possible molecular mechanism. METHODS: The recombinant ESAT6 protein was obtained from E.coli carrying esat6 gene after isopropyl ß-d-thiogalactopyranoside (IPTG) induction. BMDMs were isolated from bone marrow of mice hind legs. Cells viability was detected by Cell Counting Kit 8 (CCK8) assays. The expression levels of mRNA and proteins were detected by qPCR and Western blot, or evaluated by flow cytometry. The expression level of nitric oxide (NO) was measured with a nitric oxide indicator. RESULTS: ESAT6 could significantly induce mRNA and protein expression levels of a group of epithelioid macrophages marker molecules (EMMMs), including E-cadherin, junction plakoglobin, ZO1, desmoplakin, desmoglein3 and catenin porteins, in BMDMs. These events could be abrogated in macrophage from TLR2 deficiency mice. ESAT6 could also markedly induce iNOS/NO production that could significantly inhibit trimethylation of H3K27 in the cells. ESAT6-induced expressions of epithelioid macrophages marker molecules were significantly inhibited in the presence of H3K27 histone demethylase inhibitor GSK J1. Furthermore, ROS scavenging agent N,N'-Dimethylthiourea (DMTU) could markedly inhibit the transition induced by ESAT6 in macrophages. CONCLUSION: This study demonstrates that ESAT6 bound with TLR2 can activate iNOS/NO and ROS signalings to reduce the trimethylation of H3K27 resulting in the increment of EMMMs expression that is beneficial to the transition of macrophages into epithelioid macrophages. However, hypoxia can inhibit this transition event. This study has provided new evidence of pathogenesis of granuloma caused by Mtb and also proposed new ideas for the treatment of TB.
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Antígenos Bacterianos/metabolismo , Proteínas Bacterianas/metabolismo , Transdiferenciación Celular/fisiología , Macrófagos/metabolismo , Transducción de Señal/fisiología , Tuberculosis/metabolismo , Animales , Metilación de ADN/fisiología , Regulación hacia Abajo , Granuloma/metabolismo , Granuloma/microbiología , Granuloma/patología , Histonas , Macrófagos/patología , Ratones , Mycobacterium tuberculosis , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Tuberculosis/patologíaRESUMEN
Interfacial self-assembly has become a powerful force for regulating the amphipathy of Pickering emulsions on the oil/water interface. Herein, metal-phenolic supramolecular coatings, acting as a regulator on the oil/water interface, were fabricated on the surface of zein nanoparticles (NPs), as a consequence of which the prepared Pickering emulsions stabilized by the decorated zein NPs exhibited diverse properties, decided by different concentrations of zein, tannic acid (TA), and metal ions (Fe3+). Metal-phenolic network-decorated zein NPs named ZTFex NPs (ZTFe NPs represented zein/TA/Fe3+ NPs, and x represented different concentrations of compounds) exhibited increasing diameters of 100-110 nm. Three-phase contact angles also showed that the strong hydrophobicity of zein NPs could be decreased as a result of the formation of metal-phenolic networks. As for corresponding Pickering emulsions, the covering of TA-Fe3+ networks on zein NPs could enhance the stability of zein NP-based emulsion obviously, which might be due to the fact that ZTFex NPs revealed the ability to form strong films on the oil/water interfaces. ZTFe4 was selected as an optimal concentration because of its minimum size and excellent storage stability. Besides, it was also found that the diameter of ZTFe4-based emulsion enhanced with the increase in the oil phase. The rheological measurement results showed that both G' and Gâ³ increased with the increase of x and the oil phase. In general, our paper not only highlighted a straightforward method for the interfacial nanofabrication of solid particles but also provided a novel and potential strategy in Pickering emulsion applications.
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Hierro/química , Nanopartículas/química , Aceites/química , Fenoles/química , Agua/química , Zeína/química , Emulsiones/química , Interacciones Hidrofóbicas e Hidrofílicas , Tamaño de la Partícula , Reología , Taninos/químicaRESUMEN
OBJECTIVE: To investigate the effects of Astragalus membranaccus (As) on cardiac function and SERCA2a gene expression in left ventricular tissues of rats with chronic heart failure. METHODS: Heart failure was induced by clipping the abdominal aorta 60 male SD rats were divided into four groups: sham-operated (Sham), aortic stenosis (Model), Model + As (20 g/kg) and Model + Captopril (0.05 g/kg). The drugs were administered orally from the 13th week after surgery. Rats were examined after 12 weeks' treatment with drugs. The parameters of hemodynamics including LVSP, LVEDP, and +/- LVdp/dt(max) were measured. SERCA2a mRNA and protein expressions in left ventricular tissues were determined by half-quantitative RT-PCR and Western blot normalized to abundance of GAPDH mRNA and portein, respectively. RESULTS: LVSP and LVEDP were obviously enhanced (P < 0.01 or P < 0.001) in model rats in vivo. Both Captopril and As prevented the increase of LVSP (P < 0.05 or P < 0.01) and LVEDP (P < 0.05 or P < 0.01). RT-PCR and Western blot results demonstrated that SERCA2a gene expression was downregulated (P < 0.05) significantly in model group compared with sham group. As upregulated SERCA2a gene expression (P < 0.05), whereas Captopril had no effect on that. CONCLUSION: As can ameliorate abnormity of cardiac function, especially diastoilc function in rats with pressure overload-induced heart failure, and that may be partly related to its up-regulation of SERCA2a gene expressions in left ventricular tissues.
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Astragalus propinquus , Medicamentos Herbarios Chinos/farmacología , Insuficiencia Cardíaca/tratamiento farmacológico , Corazón/efectos de los fármacos , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico/metabolismo , Animales , Astragalus propinquus/química , Captopril/farmacología , Enfermedad Crónica , Modelos Animales de Enfermedad , Medicamentos Herbarios Chinos/química , Regulación de la Expresión Génica/efectos de los fármacos , Corazón/fisiopatología , Insuficiencia Cardíaca/metabolismo , Insuficiencia Cardíaca/fisiopatología , Ventrículos Cardíacos/efectos de los fármacos , Ventrículos Cardíacos/metabolismo , Masculino , ARN Mensajero/metabolismo , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico/genéticaRESUMEN
The effects of Cd2+ and Cu2+ at 300 mg/L on anaerobic microbial communities that degrade 2-cholorophenol (2-CP) were examined. Based on the polymerase chain reaction (PCR) of 16S rDNA, bacterial community diversity and archaeal community structure were analyzed with denaturing gradient gel electrophoresis (DGGE) and cloning, respectively. Degradation capabilities of the anaerobic microbial community were drastically abated and the degradation efficiency of 2-CP was reduced to 60% after shock by Cu2+ and Cd2+, respectively. The bacterial community structure was disturbed and the biodiversity was reduced after shock by Cu2+ and Cd2+ for 3 d. Some new metal-resistant microbes which could cope with the new condition appeared. The sequence analysis showed that there existed common Archaea species in control sludge and systems when treated with Cu2+ and Cd2+, such as Methanothrix soehngenii, Methanosaeta concilii, uncultured euryarchaeote, and so on. Both the abundance and diversity of archaeal species were altered with addition of Cd2+ and Cu2+ at high concentration. Although the abundance of the predominant archaeal species decreased with Cd2+ and Cu2+ addition for 3 d, they recovered to some extent after 10 d. The diversity of archaeal species was remarkably reduced after recovery for 10 d and the shift in archaeal composition seemed to be irreversible. The 2-CP-degradation anaerobic system was more sensitive to Cu2+ than Cd2+.
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Archaea/metabolismo , Bacterias/metabolismo , Reactores Biológicos , Cadmio/metabolismo , Clorofenoles/metabolismo , Cobre/metabolismo , Anaerobiosis , Archaea/clasificación , Archaea/genética , Bacterias/clasificación , Bacterias/genética , Reactores Biológicos/microbiología , ADN Ribosómico/análisis , ADN Ribosómico/genética , Datos de Secuencia Molecular , Filogenia , Aguas del Alcantarillado/microbiologíaRESUMEN
OBJECTIVE: To investigate the effect of astragalus (As) on calcium accumulation and SERCA2a gene expression in left ventricular tissues in rats with pressure overload-induced cardiac hypertrophy. METHOD: cardiac hypertrophy was induced by clipping the abdominal aorta in rats. Male SD rats were allocated to six groups: sham-operrated (Sham), aortic stenosis (Model), model +As-L (5 g x kg(-1) x d(-1)), model+As-M (10 g x kg(-1) x d(-1)), model+As-H (20 g x kg(-1) x d(-1)) and model + captopril (0.05 mg x kg(-1) x d(-1), a positive control). The drugs were administered orally from the 13 th week after surgery. Rats were examined after 12 week treatment with drugs. The cardiac hypertrophy was evaluated by left ventricular mass index (LVMI, left ventricular weight/ body weight). The calcium content in left ventricular tissue was measured by atomic absorption spectrometry. SERCA2a mRNA and protein expressions in left ventricular tissues were determined by half-quantitative RT-PCR and Western blot normalized to abundance of GAPDH mRNA and protein, respectively. RESULT: The increase of LVMI was dose-dependently lessened by As (P < 0.01, P < 0.001). The effect of As-H was similar to that of Captopril. As markedly attenuated calcium accumulation in myocardial tissure (P < 0.01). RT-PCR and Western blot results demonstrated that SERCA2a gene expressions were downregulated (P < 0.05) significantly in model group compared with sham group. As-H upregulated SERCA2a gene expressions (P < 0.05), whereas Captopril had no effect on that. CONCLUSION: The inhibition of As on left ventricular hypertrophy induced by pressure overload in rats may partly contribute to its attenuation of calcium accumulation and up-regulation of SERCA2a gene expressions in left ventricular tissues.
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Planta del Astrágalo/química , Calcio/metabolismo , Medicamentos Herbarios Chinos/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Hipertrofia Ventricular Izquierda/metabolismo , Miocardio/metabolismo , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico/metabolismo , Animales , Western Blotting , Medicamentos Herbarios Chinos/química , Corazón/efectos de los fármacos , Hipertrofia Ventricular Izquierda/fisiopatología , Masculino , Ratas , Ratas Sprague-Dawley , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico/genéticaRESUMEN
Diffuse axonal injury (DAI) is much common during traumatic brain injury (TBI) and is associated with high mortality and poor neurological outcome. Although many studies have been examined, there are still no reliable objective diagnostic modalities available for clinicians to make an early diagnosis of DAI. Therefore, we established a rat model of DAI, applying an integrated 1H NMR- and UPLC-Q-TOF/MS-based metabonomics approach to identify differentially changed metabolites in plasma. A total of twenty-two metabolites in the injury group were identified as differentially changed. Among them, four metabolites, glutamine, pyruvate, glycerol and phosphocholine, were identified as candidate biomarkers based on their high fold-changes and biological functions, and may play important roles in axonal injury progression in DAI. Our study not only identified several novel biomarkers that improved our understanding of the metabolic events underlying DAI, but also may provide some potential novel therapeutic targets for preventing axonal injury in DAI.
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Lesión Axonal Difusa/sangre , Metabolómica/métodos , Animales , Biomarcadores/sangre , Encéfalo/patología , Cromatografía Liquida , Lesión Axonal Difusa/patología , Modelos Animales de Enfermedad , Masculino , Espectrometría de Masas , Metaboloma , Resonancia Magnética Nuclear Biomolecular , Espectroscopía de Protones por Resonancia Magnética , Distribución Aleatoria , Ratas Sprague-DawleyRESUMEN
Delayed encephalopathy after acute carbon monoxide poisoning (DEACMP) is a difficult-to-manage neurological complication that can severely affect the life quality of patients. Although the central nervous system (CNS) injuries have been reported, the underlying molecular mechanisms are still unclear. Therefore, we established a rat model of DEACMP, applying isobaric tags for a relative and absolute quantification (iTRAQ)-based proteomics approach to identify differentially expressed proteins in cerebral tissue. A total of 170 proteins in the CO exposure groups were identified as differentially changed. Bioinformatics analysis suggested that these proteins are mainly involved in the biological processes, such as energy metabolism and many neurodegenerative diseases. Three proteins, Glial fibrillary acidic protein (GFAP), mitochondrial malate dehydrogenase (MDHM), and isocitrate dehydrogenase [NAD] subunit alpha (IDH3A), were identified as playing important roles in CNS injuries in DEACMP, and were successfully confirmed by immunohistochemistry analysis. Our study not only offers us new insights into the pathophysiological mechanisms of CNS injuries in DEACMP, but also may provide clinicians with important references in early prevention and treatment.
Asunto(s)
Encefalopatías/etiología , Encefalopatías/metabolismo , Encéfalo/metabolismo , Intoxicación por Monóxido de Carbono/metabolismo , Proteoma , Animales , Encéfalo/patología , Encefalopatías/patología , Encefalopatías/psicología , Intoxicación por Monóxido de Carbono/patología , Intoxicación por Monóxido de Carbono/psicología , Supervivencia Celular , Biología Computacional , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Proteína Ácida Fibrilar de la Glía/metabolismo , Inmunohistoquímica , Isocitrato Deshidrogenasa/metabolismo , Malato Deshidrogenasa/metabolismo , Masculino , Aprendizaje por Laberinto , Proteómica , Distribución Aleatoria , Ratas Sprague-Dawley , Memoria Espacial , Factores de TiempoRESUMEN
OBJECTIVES: To reveal whether B-myb is involved in preventing senescence of vascular endothelial cells, and if so, to identify possible mechanisms for it. MATERIALS AND METHODS: C57/BL6 male mice and primary human aortic endothelial cells (HAECs) were used. Bleomycin was applied to induce stress-related premature senescence. B-myb knockdown was achieved using an siRNA technique and cell senescence was assessed using the senescence-associated ß-galactosidase (SA-ß-gal) assay. Intracellular reactive oxygen species (ROS) production was analysed using an ROS assay kit and cell proliferation was evaluated using KFluor488 EdU kit. Capillary tube network formation was determined by Matrigel assay. Expressions of mRNA and protein levels were detected by real-time PCR and western blotting. RESULTS: B-myb expression significantly decreased, while p53 and p21 expressions increased in the aortas of aged mice. This expression pattern was also found in replicative senescent HAECs and senescent HAECs induced by bleomycin. B-myb knockdown resulted in upregulation of p22phox , ROS accumulation and cell senescence of HAECs. Downregulation of B-myb significantly inhibited cell proliferation and capillary tube network formation and activated the p53/p21 signalling pathway. Blocking ROS production or inhibiting p53 activation remarkably attenuated SA-ß-gal activity and delayed cell senescence induced by B-myb-silencing. CONCLUSION: Downregulation of B-myb induced senescence by upregulation of p22phox and activation of the ROS/p53/p21 pathway, in our vascular endothelial cells, suggesting that B-myb may be a novel candidate for regulating cell senescence to protect against endothelial senescence-related cardiovascular diseases.
Asunto(s)
Proteínas de Ciclo Celular/metabolismo , Proliferación Celular/fisiología , Células Endoteliales/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal , Transactivadores/metabolismo , Animales , Senescencia Celular/fisiología , Regulación hacia Abajo , Masculino , Ratones Endogámicos C57BL , ARN Interferente Pequeño/metabolismo , Transducción de Señal/fisiología , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
Endothelial dysfunction caused by endothelial cells senescence and chronic inflammation is tightly linked to the development of cardiovascular diseases. NLRP3 (NOD-like receptor family pyrin domain-containing3) inflammasome plays a central role in inflammatory response that is associated with diverse inflammatory diseases. This study explores the effects and possible mechanisms of NLRP3 inflammasome in endothelial cells senescence. Results show an increment of pro-inflammatory cytokine interleukin (IL) -1ß secretion and caspase-1 activation during the senescence of endothelial cells induced by bleomycin. Moreover, secreted IL-1ß promoted endothelial cells senescence through up-regulation of p53/p21 protein expression. NLRP3 inflammasome was found to mediate IL-1ß secretion through the production of ROS (reactive oxygen species) during the senescence of endothelial cells. Furthermore, the association of TXNIP (thioredoxin-interacting protein) with NLRP3 induced by ROS promoted NLRP3 inflammasome activation in senescent endothelial cells. In addition, the expressions of NLRP3 inflammasome related genes, ASC (apoptosis associated speck-like protein containing a CARD), TXNIP, cleaved caspase-1 and IL-1ß, were also increased in vitro and in vivo studies. These findings indicate that endothelial senescence could be mediated through ROS and NLRP3 inflammasome signaling pathways, suggesting a potential target for the prevention of endothelial senescence-related cardiovascular diseases.
Asunto(s)
Proteínas Portadoras/metabolismo , Células Endoteliales/citología , Células Endoteliales/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Envejecimiento/genética , Envejecimiento/metabolismo , Envejecimiento/patología , Animales , Aorta/metabolismo , Aorta/patología , Proteínas Reguladoras de la Apoptosis/antagonistas & inhibidores , Proteínas Reguladoras de la Apoptosis/genética , Proteínas Reguladoras de la Apoptosis/metabolismo , Bleomicina/toxicidad , Proteínas Adaptadoras de Señalización CARD , Proteínas Portadoras/antagonistas & inhibidores , Proteínas Portadoras/genética , Senescencia Celular/efectos de los fármacos , Senescencia Celular/fisiología , Proteínas del Citoesqueleto/antagonistas & inhibidores , Proteínas del Citoesqueleto/genética , Proteínas del Citoesqueleto/metabolismo , Células Endoteliales/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana , Humanos , Inflamasomas/metabolismo , Interleucina-1beta/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Modelos Cardiovasculares , Proteína con Dominio Pirina 3 de la Familia NLR/antagonistas & inhibidores , Proteína con Dominio Pirina 3 de la Familia NLR/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , ARN Interferente Pequeño/genética , Especies Reactivas de Oxígeno/metabolismo , Tiorredoxinas/metabolismoRESUMEN
Regulation of leukocyte recruitment is an important determinant of the host response to microbial infection. Because tissue infiltration by inflammatory cells represents a potential source of unnecessary tissue damage, the process may be controlled by modulating the expression of chemoattractants and the receptors through which they promote directed leukocyte migration. In the present report, we show that expression of the receptor for chemotactic formylated peptides (FPR1)is negatively regulated in both macrophages and neutrophils by interleukin-4 (IL-4). The reduction of FPR1 mRNA occurs rapidly in response to both IL-4 and IL-13 but endures for <4 h after the removal of IL-4. As with many other responses to IL-4 and IL-13, suppression of FPR1 expression is dependent on the activation of Stat6. The inhibitory effect exhibits relative stimulus specificity in that other Stat-activating cytokines, such as interferon-gamma (IFN-gamma), IFN-beta, and IL-10, have no effect. Using nuclear run-on analysis, the rate of FPR1 gene transcription is high but is not suppressed by IL-4. Moreover, IL-4 does not appear to alter the rate of FPR mRNA decay. Nevertheless, FPR mRNA exhibits a short half-life (< or =2 h), and this appears to be a critical feature of the ability of IL-4 to reduce expression. Taken together, the data suggest that IL-4 and IL-13 suppress the expression of FPR1 mRNA via a mechanism that operates to eliminate primary transcripts prior to maturation and depends on the constitutive instability of preexisiting mRNA.
Asunto(s)
Regulación de la Expresión Génica/fisiología , Interleucina-4/metabolismo , Macrófagos Peritoneales/metabolismo , Receptores de Formil Péptido/genética , Animales , Ratones , ARN Mensajero/metabolismo , Receptores de Formil Péptido/biosíntesis , Factor de Transcripción STAT6 , Transactivadores/metabolismoRESUMEN
Cell-cell communication depends on cytokine and growth factor network. Bound to their receptors on the surface of target cell, these glycoproteins initiate a range of intracellular events. Subsequent dissipation of receptor signaling is essential to ensure the response of the cell does not become pathogenic. The Suppressors of cytokine signaling (SOCS) proteins are a family of proteins induced to attenuate cytokine signal transduction in response to signals from a diverse range of cytokines and growth factors. Current evidence indicates that intracellular JAK-STAT (Janus kinase-signal transducer and activator of transcription) signaling not only governs cytokine-induced immunological responses but also rapidly initiates SOCS expression and its biological functions. This review focuses on current understanding of SOCS3, a member of SOCS family. SOCS3 binds to JAK, certain cytokine receptors in intracellular domain, and some signaling molecules, which results in suppressing further signaling events in the cell. Studies using conditional knockout mice have shown that SOCS3 protein is the key physiological regulator and plays an important pathological role in immune homeostasis. Dysregulation of SOCS3 functions can cause a variety of diseases, including allergy, autoimmune diseases, inflammation and cancer.