RESUMEN
Activated neutrophil-derived exosomes reportedly contribute to the proliferation of airway smooth muscle cells (ASMCs), thereby aggravating the airway wall remodeling during asthma; however, the specific mechanism remains unclear. Lipopolysaccharide (LPS)-EXO and si-CRNDE-EXO were extracted from the media of human neutrophils treated with LPS and LPS + si-CRNDE (a siRNA targets long non-coding RNA CRNDE), respectively. Human ASMCs were co-cultured with LPS-EXO or si-CRNDE-EXO, and cell viability, proliferation and migration were measured. The interplay of colorectal neoplasia differentially expressed (CRNDE), inhibitor of nuclear factor kappa B kinase subunit beta (IKKß) and nuclear receptor subfamily 2 group C member 2 (TAK1) was explored using RNA immunoprecipitation (RIP) and Co-IP assays. A mouse model of asthma was induced using ovalbumin. CRNDE was upregulated in LPS-EXO and successfully transferred from LPS-treated neutrophils to ASMCs through exosome. Mechanically, CRNDE loaded in LPS-EXO reinforced TAK1-mediated IKKß phosphorylation, thereby activating the nuclear factor kappa B (NF-κB) pathway. Functionally, silencing CRNDE in LPS-EXO, an IKKß inhibitor, and an NF-κB inhibitor all removed the upregulation of cell viability, proliferation and migration induced by LPS-EXO in ASMCs. In the end, the in vivo experiment demonstrated that CRNDE knockdown in neutrophils effectively reduced the thickness of bronchial smooth muscle in a mouse model for asthma. Activated neutrophils-derived CRNDE was transferred to ASMCs through exosomes and activated the NF-κB pathway by enhancing IKKß phosphorylation. The latter promoted the proliferation and migration of ASMCs and then contributed to airway remodeling in asthma.
Asunto(s)
Asma , Neoplasias Colorrectales , ARN Largo no Codificante , Remodelación de las Vías Aéreas (Respiratorias) , Animales , Asma/genética , Proliferación Celular/genética , Neoplasias Colorrectales/metabolismo , Modelos Animales de Enfermedad , Humanos , Quinasa I-kappa B/genética , Quinasa I-kappa B/metabolismo , Lipopolisacáridos/farmacología , Ratones , Miocitos del Músculo Liso/metabolismo , FN-kappa B/genética , FN-kappa B/metabolismo , Neutrófilos/metabolismo , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismoRESUMEN
Heart failure with preserved ejection fraction (HFpEF) is closely associated with metabolic derangement. Sodium glucose cotransporter-2 inhibitors (SGLT2i) and glucagon-like peptide-1 receptor agonists (GLP-1RA) exert anti-HFpEF effects, but the underlying mechanisms remain unclear. In this study, we explored the anti-HFpEF effects of empagliflozin and liraglutide and the underlying molecular mechanisms in a mouse model of HFpEF. This model was established by high-fat diet (HFD) feeding plus Nω-nitro-L-arginine methyl ester (L-NAME) treatment. The mice were treated with empagliflozin (20 mg·kg-1·d-1, i.g.) or liraglutide (0.3 mg·kg-1·d-1, i.p.) or their combination for 4 weeks. At the end of the experimental protocol, cardiac function was measured using ultrasound, then mice were euthanized and heart, liver, and kidney tissues were collected. Nuclei were isolated from frozen mouse ventricular tissue for single-nucleus RNA-sequencing (snRNA-seq). We showed that administration of empagliflozin or liraglutide alone or in combination significantly improved diastolic function, ameliorated cardiomyocyte hypertrophy and cardiac fibrosis, as well as exercise tolerance but no synergism was observed in the combination group. Furthermore, empagliflozin and/or liraglutide lowered body weight, improved glucose metabolism, lowered blood pressure, and improved liver and kidney function. After the withdrawal of empagliflozin or liraglutide for 1 week, these beneficial effects tended to diminish. The snRNA-seq analysis revealed a subcluster of myocytes, in which Erbb4 expression was down-regulated under HFpEF conditions, and restored by empagliflozin or liraglutide. Pseudo-time trajectory analysis and cell-to-cell communication studies confirmed that the Erbb4 pathway was a prominent pathway essential for both drug actions. In the HFpEF mouse model, both empagliflozin and liraglutide reversed Erbb4 down-regulation. In rat h9c2 cells, we showed that palmitic acid- or high glucose-induced changes in PKCα and/or ERK1/2 phosphorylation at least in part through Erbb4. Collectively, the single-cell atlas reveals the anti-HFpEF mechanism of empagliflozin and liraglutide, suggesting that Erbb4 pathway represents a new therapeutic target for HFpEF. Effects and mechanisms of action of empagliflozin and liraglutide in HFpEF mice. HFpEF was induced with a high-fat diet and L-NAME for 15 weeks, and treatment with empagliflozin and liraglutide improved the HFpEF phenotype. Single nucleus RNA sequencing (snRNA-seq) was used to reveal the underlying mechanism of action of empagliflozin and liraglutide.
Asunto(s)
Compuestos de Bencidrilo , Glucósidos , Insuficiencia Cardíaca , Liraglutida , Ratones Endogámicos C57BL , Transducción de Señal , Inhibidores del Cotransportador de Sodio-Glucosa 2 , Animales , Compuestos de Bencidrilo/farmacología , Compuestos de Bencidrilo/uso terapéutico , Glucósidos/farmacología , Glucósidos/uso terapéutico , Liraglutida/farmacología , Liraglutida/uso terapéutico , Transducción de Señal/efectos de los fármacos , Masculino , Ratones , Insuficiencia Cardíaca/tratamiento farmacológico , Insuficiencia Cardíaca/metabolismo , Inhibidores del Cotransportador de Sodio-Glucosa 2/farmacología , Inhibidores del Cotransportador de Sodio-Glucosa 2/uso terapéutico , Dieta Alta en Grasa , Volumen Sistólico/efectos de los fármacos , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Modelos Animales de EnfermedadRESUMEN
Within the context of residual cardiovascular risk in post-statin era, emerging evidence from epidemiologic and human genetic studies have demonstrated that triglyceride (TG)-rich lipoproteins and their remnants are causally related to cardiovascular risk. While, carriers of loss-of-function mutations of ApoC3 have low TG levels and are protected from cardiovascular disease (CVD). Of translational significance, siRNAs/antisense oligonucleotide (ASO) targeting ApoC3 is beneficial for patients with atherosclerotic CVD. Therefore, animal models of atherosclerosis with both hypercholesterolemia and hypertriglyceridemia are important for the discovery of novel therapeutic strategies targeting TG-lowering on top of traditional cholesterol-lowering. In this study, we constructed a novel mouse model of familial combined hyperlipidemia through inserting a human ApoC3 transgene (hApoC3-Tg) into C57BL/6 J mice and injecting a gain-of-function variant of adeno-associated virus-proprotein convertase subtilisin/kexin type 9 (AAV-PCSK9)-D377Y concurrently with high cholesterol diet (HCD) feeding for 16 weeks. In the last 10 weeks, hApoC3-Tg mice were orally treated with a combination of atorvastatin (10 mg·kg-1·d-1) and fenofibrate (100 mg·kg-1·d-1). HCD-treated hApoC3-Tg mice demonstrated elevated levels of serum TG, total cholesterol (TC) and low density lipoprotein-cholesterol (LDL-C). Oral administration of atorvastatin and fenofibrate significantly decreased the plaque sizes of en face aorta, aortic sinus and innominate artery accompanied by improved lipid profile and distribution. In summary, this novel mouse model is of considerable clinical relevance for evaluation of anti-atherosclerotic drugs by targeting both hypercholesterolemia and hypertriglyceridemia.
Asunto(s)
Aterosclerosis , Modelos Animales de Enfermedad , Hiperlipidemia Familiar Combinada , Ratones Endogámicos C57BL , Ratones Transgénicos , Animales , Aterosclerosis/tratamiento farmacológico , Humanos , Ratones , Hiperlipidemia Familiar Combinada/tratamiento farmacológico , Hiperlipidemia Familiar Combinada/genética , Apolipoproteína C-III/genética , Masculino , Proproteína Convertasa 9/genética , Proproteína Convertasa 9/metabolismo , Hipolipemiantes/uso terapéutico , Hipolipemiantes/farmacología , Triglicéridos/sangre , Dieta Alta en Grasa , Atorvastatina/uso terapéutico , Atorvastatina/farmacologíaRESUMEN
Urolithin A (UroA), a dietary phytochemical, is produced by gut bacteria from fruits rich in natural polyphenols ellagitannins (ETs). The efficiency of ETs metabolism to UroA in humans depends on gut microbiota. UroA has shown a variety of pharmacological activities. In this study we investigated the effects of UroA on atherosclerotic lesion development and stability. Apolipoprotein E-deficient (ApoE-/-) mice were fed a high-fat and high-cholesterol diet for 3 months to establish atherosclerosis model. Meanwhile the mice were administered UroA (50 mg·kg-1·d-1, i.g.). We showed that UroA administration significantly decreased diet-induced atherosclerotic lesions in brachiocephalic arteries, macrophage content in plaques, expression of endothelial adhesion molecules, intraplaque hemorrhage and size of necrotic core, while increased the expression of smooth muscle actin and the thickness of fibrous cap, implying features of plaque stabilization. The underlying mechanisms were elucidated using TNF-α-stimulated human endothelial cells. Pretreatment with UroA (10, 25, 50 µM) dose-dependently inhibited TNF-α-induced endothelial cell activation and monocyte adhesion. However, the anti-inflammatory effects of UroA in TNF-α-stimulated human umbilical vein endothelial cells (HUVECs) were independent of NF-κB p65 pathway. We conducted RNA-sequencing profiling analysis to identify the differential expression of genes (DEGs) associated with vascular function, inflammatory responses, cell adhesion and thrombosis in UroA-pretreated HUVECs. Human disease enrichment analysis revealed that the DEGs were significantly correlated with cardiovascular diseases. We demonstrated that UroA pretreatment mitigated endothelial inflammation by promoting NO production and decreasing YAP/TAZ protein expression and TEAD transcriptional activity in TNF-α-stimulated HUVECs. On the other hand, we found that UroA administration modulated the transcription and cleavage of lipogenic transcription factors SREBP1/2 in the liver to ameliorate cholesterol metabolism in ApoE-/- mice. This study provides an experimental basis for new dietary therapeutic option to prevent atherosclerosis.
RESUMEN
Urban areas experience numerous environmental challenges, among which the anthropogenic emissions of heat and carbon are two major contributors, the former is responsible for the notorious urban heat effect, the latter longterm climate changes. Moreover, the exchange of heat and carbon dioxide are closely interlinked in the built environment, and can form positive feedback loops that accelerate the degradation of urban environmental quality. Among a handful countermeasures for heat and carbon mitigation, urban irrigation is believed to be effective in cooling, yet the understanding of its impact on the co-evolution of heat and carbon emission remains obscure. In this study, we conducted multiphysics urban climate modeling for all urban areas in the contiguous United States, and evaluated the irrigation-induced cooling and carbon mitigation. Furthermore, we assessed the impact of urban irrigation on the potential heat-carbon feedback loop, with their strength of coupling quantified by an advanced causal inference method using the convergent cross mapping algorithms. It is found that the impact of urban irrigation varies vastly in geographically different cities, with its local and non-local effect unraveling distinct pathways of heat-carbon feedback mechanism.
Asunto(s)
Frío , Calor , Estados Unidos , Ciudades , Temperatura , RetroalimentaciónRESUMEN
The quantification of cross-regional interactions for the atmospheric transport processes is of crucial importance to improve the predictive capacity of climatic and environmental system modeling. The dynamic interactions in these complex systems are often nonlinear and non-separable, making conventional approaches of causal inference, such as statistical correlation or Granger causality, infeasible or ineffective. In this study, we applied an advanced approach, based on the convergent cross mapping algorithm, to detect and quantify the causal influence among different climate regions in the contiguous U.S. in response to temperature perturbations using the long-term (1901-2018) climatology of near surface air temperature record. Our results show that the directed causal network constructed by convergent cross mapping algorithm, enables us to distinguish the causal links from spurious ones rendered by statistical correlation. We also find that the Ohio Valley region, as an atmospheric convergent zone, acts as the regional gateway and mediator to the long-term thermal environments in the U.S. In addition, the temporal evolution of dynamic causality of temperature exhibits superposition of periodicities at various time scales, highlighting the impact of prominent low frequency climate variabilities such as El Niño-Southern Oscillation. The proposed method in this work will help to promote novel system-based and data-driven framework in studying the integrated environmental system dynamics.
Asunto(s)
Algoritmos , Ohio , Temperatura , Estados UnidosRESUMEN
The present study was aimed to explore the involvement of dopamine D1 receptor of the anterior cingulate cortex (ACC) in the regulation of chronic inflammatory pain-related emotion. On the first day, the rats were acclimated to the environment and the baseline indices were measured. On the second day, the rats were administered with the dopamine D1 receptor antagonist SCH-23390 or agonist SKF38393 in the ACC, and then they were subcutaneously injected with complete Freund's adjuvant (CFA, 0.08 mL) in the left hind paw to establish conditioned place avoidance (CPA) response after pairing with specific environment. On the third day, the CPA response and the firing frequency of ACC neurons were observed synchronously, and the open-field behavior, mechanical pain behavior and paw withdrawal latency (PWL) tests were also observed subsequently. In other experiments, rats were given subcutaneous injection of normal saline (NS) on the left hind paw after SCH-23390 or SKF-38393 was administered in the ACC, and then the same observations were performed. The results showed that: (1) Compared with the control group, the PWL and mechanical pain thresholds of rats injected with CFA on the left hind paw were significantly decreased (P < 0.05); (2) The residence time of rats injected with CFA in the "pain environment" and open field center was significantly shortened (P < 0.05); (3) Pre-injection of antagonist SCH-23390 in ACC (10 µg) alleviated the anxiety-like negative behavior response induced by CFA (P < 0.05) and reversed CFA-induced increases of discharge frequency of ACC neurons (P < 0.05); (4) Pre-injection of agonist SKF-38393 in the ACC (10 µg) induced CPA-like behavioral response in rats injected with NS in the left hind paw, and increased the firing frequency of ACC neurons (P < 0.05); (5) Immunofluorescence detection showed that dopamine D1 receptor and NMDA receptor were co-expressed in the same neuron. These results suggest that inhibition of dopamine D1 receptor in ACC can alleviate the negative emotional response induced by persistent pain.
Asunto(s)
Dolor Crónico , Giro del Cíngulo , 2,3,4,5-Tetrahidro-7,8-dihidroxi-1-fenil-1H-3-benzazepina/efectos adversos , Animales , Ansiedad , Hiperalgesia , Ratas , Receptores de Dopamina D1/metabolismoRESUMEN
X-linked adrenoleukodystrophy (X-ALD) is an inherited disease caused by a mutation in the adenosine 5'-triphosphate binding cassette subfamily D member 1 (ABCD1) gene encoding a peroxisomal transmembrane protein, which has various clinical manifestations and a rapid progression from initial symptoms to fatal inflammatory demyelination. Therefore, identification of early clinical symptoms and further early diagnosis as well as treatment can effectively prevent disease development. In this study, we reported the laboratory and radiographic features in a rare case of X-ALD with 3-year skin hyperpigmentation as the only manifestation. And the ABCD1 gene was sequenced for the patient and his parents by a high-throughput sequencing method. The results of laboratory examination showed adrenocortical hypofunction and increased serum concentrations of very long-chain fatty acids. Brain MRI showed no obvious abnormal signal shadow. A hemizygous mutation of c.521A>C was detected in the ABCD1 gene of the patient, and his mother has the same site heterozygous mutation. Therefore, this patient was diagnosed as "X-linked adrenoleukodystrophy". During the follow-up, adrenocortical hypothyroidism did not improve, and brain MRI showed few high-FLAIR signals in the white matter of the right radial corona and left parietal lobe, suggesting possible brain injury. X-ALD patients with only skin manifestations but no neurological abnormalities are easily neglected, but early diagnosis and early intervention are important ways to delay the progression of this disease. Therefore, genetic testing for early X-ALD is recommended in all male children patients with skin pigmentation as the sole clinical presentation and subsequent diagnosis of adrenal hypofunction.
Asunto(s)
Adrenoleucodistrofia , Hiperpigmentación , Niño , Humanos , Masculino , Adrenoleucodistrofia/diagnóstico , Adrenoleucodistrofia/genética , Adrenoleucodistrofia/complicaciones , Miembro 1 de la Subfamilia D de Transportador de Casetes de Unión al ATP/genética , Pruebas Genéticas , Hiperpigmentación/etiología , Hiperpigmentación/genética , Imagen por Resonancia Magnética/efectos adversos , Imagen por Resonancia Magnética/métodosRESUMEN
Urban greening has been as a popular and effective strategy for ameliorating urban thermal environment and air quality. Nevertheless, it remains an outstanding challenge for numerical urban models to disentangle and quantify the complex interplay between heat and carbon dynamics. In this study, we used a newly developed coupled urban canopy-carbon dynamics model to investigate the environmental co-benefits for mitigating urban heat stress as well as the reduction of carbon dioxide (CO2) emission. In particular, we evaluated the impact of specific components of urban greening, viz. fraction of the urban lawn, bare soil, tree coverage, and irrigation on heat and carbon fluxes in the built environment. The results of numerical simulations show that the expansion of urban green space, in general, leads to environmental cooling and reduced CO2 emission, albeit the efficacy varies for different vegetation types. In addition, adequate irrigation is essential to effect plant physiological functions for cooling and CO2 uptake, whereas further improvement becomes marginal with excessive irrigation. The findings of this study, along with its implications on environmental management, will help to promote sustainable urban development strategies for achieving desirable environmental co-benefits for urban residents and practitioners.
Asunto(s)
Contaminación del Aire , Ciclo del Carbono , Dióxido de Carbono/análisis , Plantas , SueloRESUMEN
Cardiac hypertrophy is a common pathological process of various cardiovascular diseases and eventually develops into heart failure. This paper was aimed to study the different pathological characteristics exhibited by different mouse strains after hypertrophy stimulation. Two mouse strains, A/J and FVB/nJ, were treated with isoproterenol (ISO) by osmotic pump to induce cardiac hypertrophy. Echocardiography was performed to monitor heart morphology and function. Mitochondria were isolated from hearts in each group, and oxidative phosphorylation function was assayed in vitro. The results showed that both strains showed a compensatory enhancement of heart contractile function after 1-week ISO treatment. The A/J mice, but not the FVB/nJ mice, developed significant cardiac hypertrophy after 3-week ISO treatment as evidenced by increases in left ventricular posterior wall thickness, heart weight/body weight ratio, cross sectional area of cardiomyocytes and cardiac hypertrophic markers. Interestingly, the heart from A/J mice contained higher mitochondrial DNA copy number compared with that from FVB/nJ mice. Functionally, the mitochondria from A/J mice displayed faster O2 consumption at state III with either complex I substrates or complex II substrate, compared with those from FVB/nJ mice. ISO treatment did not affect mitochondrial respiratory control rate (RCR), but significantly suppressed the ADP/O ratio generated from the complex II substrate in both strains. The ADP/O ratio generated from the complex I substrates in A/J mice declined by 50% after ISO treatment, whereas FVB/nJ mice were not affected. These results suggest that, compared with FVB/nJ mice, A/J mice possesses a poor integrity of mitochondrial respiratory chain that might contribute to its vulnerability to ISO-induced cardiac hypertrophy.
Asunto(s)
Cardiomegalia , Insuficiencia Cardíaca , Animales , Cardiomegalia/inducido químicamente , Isoproterenol/metabolismo , Isoproterenol/toxicidad , Ratones , Mitocondrias , Miocitos Cardíacos/metabolismoRESUMEN
Model evaluation is a critical component in the development and applications of environmental modeling systems. Conventional metrics such as Pearson product-moment correlation coefficient (r), root-mean-square error (RMSE), and mean absolute error (MAE), albeit process-based and limited to point-to-point statistical comparison, have been widely used in model evaluations. In this study, we propose a network-based toolkit for evaluation of model performance and multi-model comparisons with applications to weather prediction and climate modeling. The model outputs are topologically quantified through a range of network metrics to provide a holistic measure of system dynamics. We first use this toolkit to evaluate the performance of air temperature simulated by the Weather Research and Forecasting model with station measurements over the contiguous United States (CONUS). Results of network analysis suggest a good match between simulation and measurement, as indicated by conventional metrics (r, RMSE, and MAE) as well. The sensitivity of these network metrics is then analyzed based on CONUS station measurements with additive random errors using Monte Carlo simulations. Network metrics show more sensitive and highly nonlinear responses to increasing random errors as compared to conventional ones. Moreover, we use the new toolkit for intercomparison of the downscaled historical air temperature outputs from four global climate models. The similarity in both metrics and spatial structure highlights the capability of network analysis for capturing system dynamics in models alike. The network theory is therefore promising for evaluation and intercomparison of various environmental modeling systems with complex dynamics.
Asunto(s)
Clima , Tiempo (Meteorología) , Predicción , TemperaturaRESUMEN
The microRNA (miRNA) miR-125b is abnormally expressed in many different types of tumors, including osteosarcoma (OS). How aberrantly expressed miR-125b participates in regulating the initiation and progression of OS is still poorly understood. In the current study, we found that in OS, miR-125b can suppress the expression of MAP kinase kinase 7 (MKK7), which can dephosphorylate and inactivate signal transducer and activator of transcription 3 (STAT3). We also identified an elevated expression level of MKK7 in OS and an association between MKK7 expression and poor prognosis. Further, miR-125b inhibited OS cell proliferation and invasion by targeting and downregulating MKK7 in vitro and suppressed tumor formation in vivo. Moreover, using Western blot analysis, we preliminarily proved that the activation (phosphorylation) of STAT3 was regulated by MKK7 at the epigenetic level. MKK7 was overexpressed in OS and associated with poor clinical results. The miR-125b-MAPK-STAT3 axis may be one of the mechanisms of OS oncogenesis and a potential target for the treatment of OS.
RESUMEN
Copper nanowires (Cu NWs) are among ideal candidates for fabricating various advanced nanodevices, especially flexible electronics and transparent conductive electrodes. However, although many efforts have been made, the commercialization of Cu NWs is still difficult. Herein, we report an in situ seed-mediated two-step strategy to synthesize well-defined Cu NWs in high yield. In the first step, that is, seed formation process, most Cu ions (85%) in situ transform to nondecahedral Cu nanodots (NDs). These Cu NDs can promote the formation of decahedral multiply twinned particles (DMTPs) and the subsequent growth of Cu NWs by selectively inhibiting the spontaneous ripening of nanoparticle (NP) byproducts in the second step. The amount and quality of Cu NDs play an important role in high-yield production of Cu NWs, and the yield was successfully increased to 2.4 times higher than that of the conventional methods. Furthermore, an effective shaking-rotating purification technique was developed to fully separate Cu NWs from the final product solution. After scaling up the reaction, 50 g of high-quality Cu NWs can be produced with a uniform size and high aspect ratio at a very low material cost of $ 0.99/g. These promising results not only provide a high-yield and low-cost synthetic route but also can promote the widespread commercialization of Cu NWs in advanced nanodevices.
RESUMEN
The blind signature is widely used in cryptography applications because it can prevent the signer from gaining the original message. Owing to the unconditional security, the quantum blind signature is more advantageous than the classical one. In this paper, we propose a new provable secure quantum blind signature scheme with the nonorthogonal single-photon BB84-state and provide a new method to encode classical messages into quantum signature states. The message owner injects a randomizing factor into the original message and then strips the blind factor from the quantum blind signature signed by the blind signer. The verifier can validate the quantum signature and announce it publicly. At last, the analytical results show that the proposed scheme satisfies all of the security requirements of the blind signature: blindness, unforgeability, non-repudiation, unlinkability, and traceability. Due to there being no use of quantum entanglement states, the total feasibility and practicability of the scheme are obviously better than the previous ones.
RESUMEN
BACKGROUND: The mechanisms underlying neointima formation remain unclear. Interferon regulatory factors (IRFs), which are key innate immune regulators, play important roles in cardiometabolic diseases. However, the function of IRF4 in arterial restenosis is unknown. METHODS: IRF4 expression was first detected in human and mouse restenotic arteries. Then, the effects of IRF4 on neointima formation were evaluated with universal IRF4-deficient mouse and rat carotid artery injury models. We performed immunostaining to identify IRF4-expressing cells in the lesions. Smooth muscle cell (SMC)-specific IRF4-knockout (KO) and -transgenic (TG) mice were generated to evaluate the effects of SMC-IRF4 on neointima formation. We used microarray, bioinformatics analysis, and chromatin immunoprecipitation assay to identify the downstream signals of IRF4 and to verify the targets in vitro. We compared SMC-IRF4-KO/Krüppel-like factor 4 (KLF4)-TG mice with SMC-IRF4-KO mice and SMC-specific IRF4-TG/KLF4-KO mice with SMC-specific IRF4-TG mice to investigate whether the effect of IRF4 on neointima formation is KLF4-dependent. The effect of IRF4 on SMC phenotype switching was also evaluated. RESULTS: IRF4 expression in both the human and mouse restenotic arteries is eventually downregulated. Universal IRF4 ablation potentiates neointima formation in both mice and rats. Immunostaining indicated that IRF4 was expressed primarily in SMCs in restenotic arteries. After injury, SMC-IRF4-KO mice developed a thicker neointima than control mice. This change was accompanied by increased SMC proliferation and migration. However, SMC-specific IRF4-TG mice exhibited the opposite phenotype, demonstrating that IRF4 exerts protective effects against neointima formation. The mechanistic study indicated that IRF4 promotes KLF4 expression by directly binding to its promoter. Genetic overexpression of KLF4 in SMCs largely reversed the neointima-promoting effect of IRF4 ablation, whereas ablation of KLF4 abolished the protective function of IRF4, indicating that the protective effects of IRF4 against neointima formation are KLF4-dependent. In addition, IRF4 promoted SMC dedifferentiation. CONCLUSIONS: IRF4 protects arteries against neointima formation by promoting the expression of KLF4 by directly binding to its promoter. Our findings suggest that this previously undiscovered IRF4-KLF4 axis plays a key role in vasculoproliferative pathology and may be a promising therapeutic target for the treatment of arterial restenosis.
Asunto(s)
Regulación de la Expresión Génica , Factores Reguladores del Interferón , Factores de Transcripción de Tipo Kruppel , Músculo Liso Vascular , Neointima , Animales , Humanos , Factores Reguladores del Interferón/genética , Factores Reguladores del Interferón/metabolismo , Factor 4 Similar a Kruppel , Factores de Transcripción de Tipo Kruppel/genética , Factores de Transcripción de Tipo Kruppel/metabolismo , Ratones , Ratones Noqueados , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patología , Neointima/genética , Neointima/metabolismo , Neointima/patología , Análisis de Secuencia por Matrices de Oligonucleótidos , RatasRESUMEN
BACKGROUND/AIMS: Arctigenin (ATG) has been shown to possess anti-inflammatory, immunemodulatory, anti-viral, anti-microbial, anti-carcinogenic, vasodilatory and anti-platelet aggregation properties. However, the protective role of ATG in prevention of arrhythmias induced by myocardial ischemia/reperfusion is unknown. The aim of this study was to investigate the anti-arrhythmia effect of ATG in an ischemia/reperfusion injured rat heart model and explore the related mechanisms. METHODS: Rats were randomly exposed to sham operation, myocardial ischemia/ reperfusion (MI/R) alone, ATG+ MI/R, pretreated with ATG in low (12.5 mg/kg/day), medium (50 mg/kg/day) and high dose (200 mg/kg/day), respectively. Ventricular arrhythmias were assessed. The activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and the level of malondialdehyde (MDA) in myocardial tissue were determined by chemical analysis. RESULTS: Compared to MI/R, rats pretreated with ATG in doses of 50 mg/kg/day and 200 mg/kg/day showed significantly reduced incidence and duration of ventricular fibrillation, ventricular tachycardia and ventricular ectopic beat (VEB), and decreased the arrhythmia score during the 30-min ischemia. Incidence and duration of ventricular tachycardia, infarction size and arrhythmia scores in these groups were significantly decreased during the 120-min reperfusion. No ventricular fibrillation occurred during the period of reperfusion. Rats pretreated with ATG in doses of 50 mg/kg/day and 200 mg/kg/ day markedly enhanced the activities of antioxidant enzymes SOD and GSH-Px, reduced the level of MDA. No differences were observed between the group pretreated with a low dose of ATG and the sham group. Administration of ATG significantly increased the expression of antioxidant stress protein Nrf2, Trx1 and Nox1. CONCLUSION: Our data suggested that ATG plays anti-arrhythmia role in ischemia/reperfusion injury, which is probably associated with attenuating oxidative stress by Nrf2 signaling pathway.
Asunto(s)
Arritmias Cardíacas/prevención & control , Furanos/farmacología , Lignanos/farmacología , Estrés Oxidativo/efectos de los fármacos , Animales , Arritmias Cardíacas/etiología , Arritmias Cardíacas/metabolismo , Modelos Animales de Enfermedad , Furanos/uso terapéutico , Glutatión Peroxidasa/metabolismo , Lignanos/uso terapéutico , Masculino , Malondialdehído/metabolismo , Infarto del Miocardio/patología , Miocardio/metabolismo , NADPH Oxidasa 1/metabolismo , Factor 2 Relacionado con NF-E2/metabolismo , Ratas , Ratas Sprague-Dawley , Daño por Reperfusión/complicaciones , Superóxido Dismutasa/metabolismo , Tiorredoxinas/metabolismoRESUMEN
Chronic intermittent hypoxia (CIH), the hallmark of obstructive sleep apnea syndrome (OSAS), has been reported to play a key role in the development of OSAS-associated cardiovascular diseases including cardiac remodeling. RhoA/Rho-kinase (ROCK) pathway has also been implicated in myocardial remodeling, but the exact mechanisms are not fully elucidated. This study's purpose is to investigate the influence of fasudil, a selective ROCK inhibitor, on CIH-induced left ventricular remodeling in rats and its possible mechanisms. Adult male Sprague-Dawley rats suffered from CIH or normoxia stimulus and were intervened with vehicle or fasudil (10 mg·kg·d, intraperitoneal injection) for 6 weeks. In this study, treatment with fasudil significantly reversed intermittent hypoxia-induced histopathological transformations and ultrastructural changes in rat myocardium. Moreover, fasudil downregulated the protein levels of RhoA and phosphorylation of myosin phosphatase targeting subunit-1 (MYPT1), thus effectively inhibited the activation of RhoA/ROCK signaling pathway. Simultaneously, activity of nuclear factor (NF)-kB was suppressed by fasudil, which was accompanied by reduced NF-kB downstream inflammatory genes including interleukin-6, tumor necrosis factor-a and monocyte chemotactic protein-1, and apoptosis. These results suggest that fasudil attenuates myocardial remodeling in CIH rats, at least partly by suppressing activation of NF-kB. Inhibition of the RhoA/ROCK pathway could become an important therapeutic target in the prevention of OSAS-related cardiomyopathy.
Asunto(s)
Apoptosis/fisiología , Hipoxia/metabolismo , Miocardio/metabolismo , Inhibidores de Proteínas Quinasas/farmacología , Remodelación Ventricular/fisiología , Quinasas Asociadas a rho/metabolismo , 1-(5-Isoquinolinesulfonil)-2-Metilpiperazina/análogos & derivados , 1-(5-Isoquinolinesulfonil)-2-Metilpiperazina/farmacología , 1-(5-Isoquinolinesulfonil)-2-Metilpiperazina/uso terapéutico , Animales , Apoptosis/efectos de los fármacos , Hipoxia/tratamiento farmacológico , Hipoxia/patología , Inflamación/tratamiento farmacológico , Inflamación/metabolismo , Inflamación/patología , Masculino , Miocardio/patología , Inhibidores de Proteínas Quinasas/uso terapéutico , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Función Ventricular Izquierda/efectos de los fármacos , Función Ventricular Izquierda/fisiología , Remodelación Ventricular/efectos de los fármacos , Quinasas Asociadas a rho/antagonistas & inhibidoresRESUMEN
BACKGROUND Osteomyelitis is one of the refractory diseases encountered in orthopedics, while Staphylococcus aureus (S. aureus) is the most common causative organism in osteomyelitis. However, the precise mechanisms underlying the bone loss caused by S. aureus infection have not been well defined. Here, we investigated the effect of S. aureus on osteoclast differentiation and the probable molecular mechanism. MATERIAL AND METHODS RAW 264.7 cells were treated for 5 days with live S. aureus, inactivated S. aureus, and S. aureus filtrate. Then, the formation of osteoclast-like cells and resorption pits was observed, and the expression of osteoclast-specific genes (TRAP, MMP-9, cathepsin K, CTR and Atp6v0d2) was detected by real-time PCR. Moreover, key proteins in the signaling pathway associated with osteoclast differentiation were detected with Western blot. RESULTS The data showed that live S. aureus, inactivated S. aureus, and S. aureus filtrate induced osteoclast formation, promoted bone resorption, and increased the expression of osteoclast-specific genes in a dose-dependent manner in the absence RANKL. In addition, we found that the S. aureus-induced osteoclastogenesis was related to the degradation of IκB-a, phosphorylation of NF-κB p65, and increased expression of NFATc1. Thus, we used JSH-23 to inhibit NF-κB transcriptional activity. The effect of the S. aureus-induced osteoclastogenesis and the expression of osteoclast-specific genes and NFATc1 were inhibited, which indicated that the NF-κB signaling pathway plays a role in S. aureus-induced osteoclastogenesis. CONCLUSIONS This study demonstrated that S. aureus induces osteoclastogenesis through its cell wall compound and secretion of small soluble molecules, and the NF-κB signaling pathway plays a role in this process.
Asunto(s)
FN-kappa B/fisiología , Osteogénesis/efectos de los fármacos , Staphylococcus aureus/patogenicidad , Animales , Resorción Ósea/metabolismo , Resorción Ósea/microbiología , Diferenciación Celular/fisiología , Regulación de la Expresión Génica/genética , Macrófagos/metabolismo , Ratones , Proteínas Quinasas Activadas por Mitógenos/metabolismo , FN-kappa B/inmunología , FN-kappa B/metabolismo , Subunidad p52 de NF-kappa B/fisiología , Factores de Transcripción NFATC , Osteoclastos/metabolismo , Osteoclastos/microbiología , Osteogénesis/inmunología , Osteogénesis/fisiología , Osteomielitis/microbiología , Células RAW 264.7 , Transducción de Señal/efectos de los fármacos , Staphylococcus aureus/metabolismo , Factor de Transcripción ReIA/metabolismoRESUMEN
Graphene-based composites have gained great attention in the field of gas sensor fabrication due to their higher surface area with additional functional groups. Decorating one-dimensional (1D) semiconductor nanomaterials on graphene also show potential benefits in gas sensing applications. Here we demonstrate the one-pot and low cost synthesis of W18O49 NWs/rGO composites with different amount of reduced graphene oxide (rGO) which show excellent gas-sensing properties towards toluene and strong dependence on their chemical composition. As compared to pure W18O49 NWs, an improved gas sensing response (2.8 times higher) was achieved in case of W18O49 NWs composite with 0.5 wt. % rGO. Promisingly, this strategy can be extended to prepare other nanowire based composites with excellent gas-sensing performance.
RESUMEN
Circulating tumor cells (CTCs) are free tumor cells shed from tumor site and enter into blood circulation. CTCs represent a reliable source of tumor cells for the molecular characteristics of the original tumor. However, the extraordinary rarity of CTCs makes the subsequent molecular and functional analysis technically challenging. Here, we describe a one-step microfludics-based immunomagnetic isolation method to isolate CTCs directly from the whole blood of lung adenocarcinoma patients. This method avoids harsh sample preparation and enrichment steps, and therefore preserves the viability (>90%) of CTCs during the in vitro isolation. The isolated CTCs are enriched in small volume (80 µL) and cultured ex vivo that leads to successful ex vivo expansion. The expanded CTCs can be frozen and thawed, which shows cell line property. Genetic sequencing on EGFRãKRASãPIK3CAãTP53 and BRAF and metabolic assay (2-NBDG) are utilized to characterize the expanded CTCs. Our results demostrated that this method is suitable for ex vivo expansion of CTCs facilitates. The genomic, proteomic and metabolic analyses of CTCs have guiding significance in tumor precise treatment.