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OBJECTIVE: Selenium (Se) is a key part of the body's oxidation defence system. However, it is unclear whether Se affects the development of aortic aneurysm (AA). An animal experiment was conducted to clarify the role of Se in AA development. METHODS: C57BL/6N male mice were fed with a Se deficient (Se-D, < 0.05 mg/kg), Se adequate (Se-A, 0.2 mg/kg), or Se supplemented (Se-S, 1 mg/kg) diet for 8 weeks. Subsequently, an AA murine model (Se-D, n = 11; Se-A, n = 12; Se-S, n = 15) was established using angiotensin II (Ang II, 1 mg/kg/min) for four weeks plus ß-aminopropionitrile (BAPN, 1 mg/mL) for the first two weeks. Saline replaced Ang II, and BAPN was removed during the modelling process for sham mice (Se-A, n = 9). To determine whether Se deficiency promoted aortic dilation via matrix metalloproteinase-2 (MMP-2), the non-specific MMP inhibitor doxycycline (Dox, 100 mg/kg/day) was given to Se-D AA mice (n = 7) for two weeks. RESULTS: The maximum aortic diameter in Se-D AA model mice was significantly increased compared with Se-A AA model mice. MMP-2 expression and activity in the aortic media of Se-D AA model mice was significantly increased compared with Se-A AA model mice. A large number of vascular smooth muscle cells (VSMCs) were found aggregating in the media of the non-dilated aorta of Se-D AA model mice, which was completely inhibited by Dox. The percentage of VSMCs in aortic media of Se-D AA model mice was significantly higher than in Se-A AA model mice. The maximum aortic diameter and occurrence rate of AA in Se-D AA model mice with Dox were significantly reduced compared with Se-D AA model mice. CONCLUSION: Se deficiency promoted dilatation of the aorta in AA model mice by increasing expression and activity of VSMC derived MMP-2, causing abnormal aggregation and proliferation of VSMCs in aortic media.
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Aneurisma de la Aorta , Selenio , Masculino , Ratones , Animales , Metaloproteinasa 2 de la Matriz/metabolismo , Músculo Liso Vascular/metabolismo , Dilatación , Selenio/farmacología , Selenio/metabolismo , Aminopropionitrilo/farmacología , Ratones Endogámicos C57BL , Aorta/metabolismo , Modelos Animales de Enfermedad , Miocitos del Músculo Liso/metabolismoRESUMEN
Candida albicans, one of the most prevalent conditional pathogenic fungi, can cause local superficial infections and lethal systemic infections, especially in the immunocompromised population. Secretory immunoglobulin A (sIgA) is an important immune protein regulating the pathogenicity of C. albicans. However, the actions and mechanisms that sIgA exerts directly against C. albicans are still unclear. Here, we investigated that sIgA directs against C. albicans hyphal growth and virulence to oral epithelial cells. Our results indicated that sIgA significantly inhibited C. albicans hyphal growth, adhesion, and damage to oral epithelial cells compared with IgG. According to the transcriptome and RT-PCR analysis, sIgA significantly affected the ergosterol biosynthesis pathway. Furthermore, sIgA significantly reduced the ergosterol levels, while the addition of exogenous ergosterol restored C. albicans hyphal growth and adhesion to oral epithelial cells, indicating that sIgA suppressed the growth of hyphae and the pathogenicity of C. albicans by reducing its ergosterol levels. By employing the key genes mutants (erg11Δ/Δ, erg3Δ/Δ, and erg3Δ/Δ erg11Δ/Δ) from the ergosterol pathway, sIgA lost the hyphal inhibition on these mutants, while sIgA also reduced the inhibitory effects of erg11Δ/Δ and erg3Δ/Δ and lost the inhibition of erg3Δ/Δ erg11Δ/Δ on the adhesion to oral epithelial cells, further proving the hyphal repression of sIgA through the ergosterol pathway. We demonstrated for the first time that sIgA inhibited C. albicans hyphal development and virulence by affecting ergosterol biosynthesis and suggest that ergosterol is a crucial regulator of C. albicans-host cell interactions. KEY POINTS: ⢠sIgA repressed C. albicans hyphal growth ⢠sIgA inhibited C. albicans virulence to host cells ⢠sIgA affected C. albicans hyphae and virulence by reducing its ergosterol levels.
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Candida albicans , Células Epiteliales , Virulencia , Candida albicans/genética , Ergosterol , Inmunoglobulina A SecretoraRESUMEN
Recent advances in intratracheal delivery strategies have sparked considerable biomedical interest in developing this promising approach for lung cancer diagnosis and treatment. However, there are very few relevant studies on the behavior and mechanism of imaging nanoparticles (NPs) after intratracheal delivery. Here, we found that nanosized perfluoro-15-crown-5-ether (PFCE NPs, â¼200 nm) exhibite significant 19F-MRI signal-to-noise ratio (SNR) enhancement than perfluorooctyl bromide (PFOB NPs) up to day 7 after intratracheal delivery. Alveolar macrophages (AMs) engulf PFCE NPs, become PFCE NPs-laden AMs, and then migrate into the tumor margin, resulting in increased tumor PFCE concentration and 19F-MRI signals. AMs-mediated translocation of PFCE NPs to lung draning lymph nodes (dLNs) decreases the background PFCE concentration. Our results shed light on the dynamic AMs-mediated translocation of intratracheally delivered PFC NPs for effective lung tumor visualization and reveal a pathway to develop and promote the clinical translation of an intratracheal delivery-based imaging strategy.
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Fluorocarburos , Neoplasias Pulmonares , Nanopartículas , Humanos , Macrófagos Alveolares , Imagen por Resonancia Magnética/métodos , Neoplasias Pulmonares/tratamiento farmacológicoRESUMEN
The Arabidopsis WRKY11 (AtWRKY11) protein is an important transcription factor involved in plant response to biotic and abiotic stresses. Its DNA-binding domain specifically binds to gene promoter regions harboring the W-box consensus motif. Herein we report the high-resolution structure of the AtWRKY11 DNA-binding domain (DBD) determined by solution NMR spectroscopy. The results show that AtWRKY11-DBD adopts an all-ß fold comprising five ß-strands packed in an antiparallel topology, stabilized by a zinc-finger motif. Structural comparison reveals that the long ß1-ß2 loop shows the highest structural variation from other available WRKY domain structures. Moreover, this loop was further found to contribute to the binding between AtWRKY11-DBD and W-box DNA. Our current study provides atomic-level structural basis for further understanding the structure-function relationship of plant WRKY proteins.
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Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Unión al ADN/metabolismo , Secuencia de Aminoácidos , Factores de Transcripción/metabolismo , Proteínas de Plantas/metabolismo , ADN , Regulación de la Expresión Génica de las Plantas , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismoRESUMEN
BACKGROUND: The accumulation and aggregation of α-synuclein (α-Syn) are characteristic of Parkinson's disease (PD). Epidemiological evidence indicates that hyperlipidemia is associated with an increased risk of PD. The levels of 27-hydroxycholesterol (27-OHC), a cholesterol oxidation derivative, are increased in the brain and cerebrospinal fluid of patients with PD. However, whether 27-OHC plays a role in α-Syn aggregation and propagation remains elusive. OBJECTIVE: The aim of this study was to determine whether 27-OHC regulates α-Syn aggregation and propagation. METHODS: Purified recombinant α-Syn, neuronal cultures, and α-Syn fibril-injected mouse model of PD were treated with 27-OHC. In addition, CYP27A1 knockout mice were used to investigate the effect of lowering 27-OHC on α-Syn pathology in vivo. RESULTS: 27-OHC accelerates the aggregation of α-Syn and enhances the seeding activity of α-Syn fibrils. Furthermore, the 27-OHC-modified α-Syn fibrils localize to the mitochondria and induce mitochondrial dysfunction and neurotoxicity. Injection of 27-OHC-modified α-Syn fibrils induces enhanced spread of α-Syn pathology and dopaminergic neurodegeneration compared with pure α-Syn fibrils. Similarly, subcutaneous administration of 27-OHC facilitates the seeding of α-Syn pathology. Genetic deletion of cytochrome P450 27A1 (CYP27A1), the enzyme that converts cholesterol to 27-OHC, ameliorates the spread of pathologic α-Syn, degeneration of the nigrostriatal dopaminergic pathway, and motor impairments. These results indicate that the cholesterol metabolite 27-OHC plays an important role in the pathogenesis of PD. CONCLUSIONS: 27-OHC promotes the aggregation and spread of α-Syn. Strategies aimed at inhibiting the CYP27A1-27-OHC axis may hold promise as a disease-modifying therapy to halt the progression of α-Syn pathology in PD. © 2023 International Parkinson and Movement Disorder Society.
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Enfermedad de Parkinson , Humanos , Ratones , Animales , Enfermedad de Parkinson/genética , alfa-Sinucleína/genética , alfa-Sinucleína/metabolismo , Hidroxicolesteroles/farmacología , ColesterolRESUMEN
Candida albicans is the main conditional pathogenic fungus among the human microbiome. Extracellular vesicles (EVs) secreted by C. albicans are important for its pathogenesis. However, the effects and mechanisms of EVs on C. albicans own growth are not clear. Here, we isolated EVs from C. albicans cells grown in four culture media, including RPMI 1640, DMEM, YPD, and YNB, and measured their effects on the own growth of C. albicans in these media. All the C. albicans EVs from the four media could promote the growth of C. albicans in RPMI 1640 and DMEM media, but had no effects in YPD and YNB media, indicating that the effects of EVs on C. albicans growth were dependent on some media contents. By comparing the media contents and transcriptome analysis, arginine was identified as the key factor for the growth promotion of C. albicans EVs. EVs activated the L-arginine/nitric oxide pathway to promote the growth of C. albicans through that EVs increased the NO levels and upregulated the expression of NO dioxygenase gene YHB1 to reduce the intracellular reactive oxygen species (ROS) and cell apoptosis. During the host cell infections, C. albicans EVs synergistically enhanced the destructive effects of C. albicans to host cells, including RAW264.7, HOK, TR146, and HGEC, suggesting that the growth promotion by EVs enhanced the pathogenesis of C. albicans. Our results demonstrated the important roles of EVs on C. albicans own growth for the first time and highlight its synergism with C. albicans to increase the pathogenesis. KEY POINTS: ⢠C. albicans extracellular vesicles (EVs) promoted its own growth. ⢠EVs activated the l-arginine/NO pathway to reduce ROS and apoptosis of C. albicans. ⢠EVs enhanced the damage to the host cell caused by C. albicans.
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Candida albicans , Vesículas Extracelulares , Humanos , Óxido Nítrico/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Vesículas Extracelulares/metabolismo , Arginina/metabolismoRESUMEN
Cardiac shockwave therapy (CSWT) is a noninvasive treatment for patients with refractory angina or myocardial ischemia. This study aims to evaluate the potential beneficial effect and safety of CSWT in patients with severe coronary artery disease (CAD) who have undergone coronary artery bypass grafting (CABG).This was a single-arm prospective cohort study. A total of 30 patients with severe CAD who were not suitable for coronary revascularization and who had undergone CABG were enrolled. All patients received CSWT for nine sessions. Evaluation was performed before and after CSWT, including the Canadian Cardiovascular Society (CCS) classification, New York Heart Association (NYHA) classification, 6-minute walk test (6MWT), Seattle Angina Questionnaire (SAQ) score, nitroglycerin dosage, echocardiography, myocardial perfusion imaging (MPI), and safety parameters. All patients were followed up at both 1 month and 9 months after CSWT.After treatment, CSWT significantly improved CCS classification (P < 0.05), NYHA classification (P < 0.05), nitroglycerin dosage (P < 0.001), and 6MWT (P < 0.05) at 1 month and 9 months after CSWT. SAQ score (P < 0.05) and left ventricular ejection fraction (LVEF; P = 0.037) by echocardiography significantly improved at 1 month after CSWT. Significant decreases in summed stress score (SSS), summed difference score (SDS), ischemic area stress, and ischemic area difference by MPI were observed at 1 month and 9 months after CSWT (P < 0.01). There were no changes in safety parameters before and after CSWT.CSWT may have a beneficial effect on improving myocardial perfusion, clinical symptoms, exertional capacity, and quality of life and is a safe alternative treatment for patients with severe CAD who have undergone CABG.
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Enfermedad de la Arteria Coronaria , Ondas de Choque de Alta Energía , Humanos , Enfermedad de la Arteria Coronaria/cirugía , Enfermedad de la Arteria Coronaria/diagnóstico , Nitroglicerina , Ondas de Choque de Alta Energía/uso terapéutico , Volumen Sistólico , Estudios Prospectivos , Calidad de Vida , Resultado del Tratamiento , Función Ventricular Izquierda , Canadá , Puente de Arteria CoronariaRESUMEN
BACKGROUND: Immune abnormalities and inflammatory responses play critical roles in progression of hypertension. Basic studies have confirmed that Th17 cell and related cytokines are important in promoting hypertension-mediated organ damage, but few clinical evidences have been published. Therefore, our study aimed to investigate the relationship between Th17 cell and its related cytokines and hypertension-mediated organ damage in human. METHODS: This study enrolled 179 patients with hypertension (including 92 with hypertension-mediated organ damage and 87 without hypertension-mediated organ damage) and 63 healthy participants. The proportion of Th17 cells in peripheral blood mononuclear cells was measured by flow cytometry. The concentrations of interleukin-17 and interleukin-23 were detected by enzyme-linked immunosorbent assay. Real time-polymerase chain reaction was used to detect the mRNA expression levels of interleukin-17, retinoic acid-related orphan receptor (ROR) γt and signal transducer and activator of transcription-3 (STAT-3). RESULTS: The proportion of Th17 cells, the concentration of interleukin-17 and interleukin-23 and the mRNA expression levels of interleukin-17, retinoic acid-related orphan receptor γt and signal transducer and activator of transcription-3 were significantly increased in hypertension-mediated organ damage group compared with those in non-hypertension-mediated organ damage group and control group (P < 0.005). CONCLUSION: Th17 cells and their associated cytokines may be involved in hypertension-mediated organ damage formation and may be able to serve as new biomarkers of hypertension-mediated organ damage and potential therapeutic targets.
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Hipertensión , Células Th17 , Estudios de Casos y Controles , Citocinas/metabolismo , Humanos , Hipertensión/diagnóstico , Hipertensión/genética , Hipertensión/metabolismo , Interleucina-17/genética , Interleucina-23/genética , Leucocitos Mononucleares/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Linfocitos T Reguladores/metabolismo , Células Th17/metabolismo , Tretinoina/metabolismoRESUMEN
Human Y-box binding protein 1 (YB-1) is a multifunctional protein and overexpressed in many types of cancer. It specifically recognizes DNA/RNA through a cold shock domain (CSD) and regulates nucleic acid metabolism. The C-terminal extension of CSD and the phosphorylation of S102 are indispensable for YB-1 function. Until now, the roles of the C-terminal extension and phosphorylation in gene transcription and translation are still largely unknown. Here, we solved the structure of human YB-1 CSD with a C-terminal extension sequence (CSDex). The structure reveals that the extension interacts with several residues in the conventional CSD and adopts a rigid structure instead of being disordered. Either deletion of this extension or phosphorylation of S102 destabilizes the protein and results in partial unfolding. Structural characterization of CSDex in complex with a ssDNA heptamer shows that all the seven nucleotides are involved in DNA-protein interactions and the C-terminal extension provides a unique DNA binding site. Our DNA-binding study indicates that CSDex can recognize more DNA sequences than previously thought and the phosphorylation reduces its binding to ssDNA dramatically. Our results suggest that gene transcription and translation can be regulated by changing the affinity of CSDex binding to DNA and RNA through phosphorylation, respectively.
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Respuesta al Choque por Frío/genética , ADN/genética , ARN/genética , Proteína 1 de Unión a la Caja Y/genética , Secuencia de Aminoácidos , Sitios de Unión/genética , ADN de Cadena Simple/genética , Proteínas de Unión al ADN/genética , Humanos , Fosforilación/genética , Dominios Proteicos/genética , Proteínas de Unión al ARN/genéticaRESUMEN
The coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been raging across the world for over two years, but the daily reported numbers of new infections and deaths are still increasing. The newly identified Omicron variant has significant changes in its transmissibility and pathogenicity due to the multiple mutations in the spike protein, posing new challenges to the global public health. World Health Organization has categorized Omicron as a variant of concern (VOC). The spread of SARS-CoV-2 and its variants has caused disruptions to the dental practice worldwide. During the course of diagnosis and treatment of dental care, face-to-face communication at close quarters, droplets, aerosols, and exposure to saliva and blood increase the risks of SARS-CoV-2 transmission. The emergence of new variants, especially the Omicron variant, has formed new challenges to dental healthcare provision. In addition, oral tissues, including the tongue and oral mucosa, can overexpress the angiotensin converting enzyme 2 (ACE2), which is also the binding receptors of SARS-CoV-2. As a result, the oral cavity is one of the target sites of SARS-CoV-2 infection. SARS-CoV-2 infection in oral cavity may cause different oral complications, such as loss of taste. However, there are few reports about Omicron and the other variants of SARS-CoV-2 and their impacts on dental healthcare provision. Herein we made an overview of the Omicron variant and its characteristics, including its pathogenicity and immune evasion, and its potential impact on dental practice. We also proposed some control measures with the aim of reducing the possible transmission of SARS-CoV-2 and its variants during dental care.
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COVID-19 , SARS-CoV-2 , Humanos , Mutación , Pandemias , SARS-CoV-2/genéticaRESUMEN
BACKGROUND: MicroRNAs (miRNAs) and long non-coding RNAs (lncRNAs) could interact with each other to play a vital role in the pathogenesis of cancers. We aimed to examine the expression profile, clinical significance and regulatory relationship of miR-130a-3p and its predicted interactive lncRNA in clear cell renal cell carcinoma (ccRCC). METHODS: Bioinformatics analysis was used to predict lncRNAs binding with miR-130a-3p. qRT-PCR was employed to detect the expression levels of miR-130a-3p and the miRNA-targeted lncRNA, and their clinical values in ccRCC were clarified. The lncRNA sponge potential of miR-130a-3p was assessed through dual-luciferase reporter assay and the biological effects of them were observed. RESULTS: Colon cancer associated transcript 1 (CCAT1) directly interacted with miR-130a-3p and negatively regulated miR-130a-3p expression. CCAT1 was upregulated and miR-130a-3p was downregulated in ccRCC cell line and tissues (all P < 0.05). High CCAT1 and low miR-130a-3p expression was correlated with larger tumor size and advanced TNM stage in ccRCC patients. High CCAT1 level suggested a poor survival prognosis. There was a negative association between CCAT1 and miR-130a-3p expression (r = - 0.373, P = 0.010). MiR-130a-3p mimic and si-CCAT1 inhibited ccRCC cell proliferation and invasion, and induced apoptosis. CONCLUSIONS: CCAT1/miR-130a-3p axis may have potential to serve as a novel diagnostic and prognostic target of ccRCC patients.
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BACKGROUND: Our study aimed to explore the expression and the biological role of lysine-specific demethylase 2A (KDM2A) in clear cell renal cell carcinoma (ccRCC). METHODS: In vitro, KDM2A expression was measured by qRT-PCR and western blot. A total of 50 patients with ccRCC were included, and KDM2A expression in ccRCC tissues was assessed by qRT-PCR and immunohistochemistry. The effects of KDM2A expression on cell biological behavior were examined by cell counting kit-8 assay, transwell assay and flow cytometry, respectively. The prognostic value of KDM2A in ccRCC was evaluated by Kaplan-Meier method. RESULTS: The KDM2A expression was significantly upregulated in ccRCC cell line (P < 0.05). Compared with para cancer tissues, ccRCC samples showed a higher KMD2A mRNA level and a larger proportion of high KDM2A expression (all P < 0.05). High KDM2A mRNA expression was more likely to occur in ccRCC tissues with tumor size > 7 cm (P = 0.005) and T3-T4 stage (P = 0.047). Knockdown of KDM2A significantly suppressed the proliferation and invasion, and promoted the apoptosis of ccRCC cells (all P < 0.05). Moreover, Kaplan-Meier survival analysis revealed that higher level of KDM2A expression in ccRCC patients was associated with lower survival rate (P = 0.004). CONCLUSIONS: Our findings demonstrated a vital role of KDM2A in the pathogenesis of ccRCC, which provides a new perspective to understand the underlying molecular mechanisms in ccRCC.
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Carcinoma de Células Renales/genética , Proteínas F-Box/genética , Regulación Neoplásica de la Expresión Génica , Histona Demetilasas con Dominio de Jumonji/genética , Neoplasias Renales/genética , Apoptosis , Carcinoma de Células Renales/metabolismo , Carcinoma de Células Renales/mortalidad , Línea Celular Tumoral , Proteínas F-Box/metabolismo , Femenino , Humanos , Técnicas In Vitro , Histona Demetilasas con Dominio de Jumonji/metabolismo , Estimación de Kaplan-Meier , Neoplasias Renales/metabolismo , Neoplasias Renales/mortalidad , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Pronóstico , ARN Mensajero/metabolismo , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Detoxifying enzyme mRNAs are potentially useful stress biomarkers. Glutathione S-transferase (GST) metabolises lipophilic organic contaminants and mitigates oxidative damage caused by environmental pollutants. Herein, 12 Chironomus kiiensis GSTs (CkGSTs1-6, CkGSTt1-2, CkGSTd1-2, CkGSTm1-2) were cloned and grouped into sigma, theta, delta and microsomal subclasses. Open reading frames (450-699 bp) encode 170-232 amino acid proteins with predicted molecular masses of 17.31-26.84 kDa and isoelectric points from 4.94 to 9.58. All 12 GSTs were expressed during all tested developmental stages, and 11 displayed higher expression in fourth-instar larvae than eggs. GST activity after 24 h of phenol exposure was used to estimate environmental phenol contamination. After exposure to sublethal concentrations of phenol for 48 h, expression and activity of CkGSTs were inhibited in C. kiiensis larvae. Expression of CkGSTd1-2 and CkGSTs1-2 varied with phenol concentration, indicating potential use as biomarkers for monitoring environmental phenol contamination.
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Chironomidae/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Glutatión Transferasa/genética , Proteínas de Insectos/genética , Fenol/toxicidad , Contaminantes Químicos del Agua/toxicidad , Animales , Chironomidae/enzimología , Chironomidae/genética , Chironomidae/crecimiento & desarrollo , Glutatión Transferasa/metabolismo , Proteínas de Insectos/metabolismo , Larva/efectos de los fármacos , Larva/enzimología , Larva/genética , Larva/crecimiento & desarrollo , Óvulo/efectos de los fármacos , Óvulo/enzimología , Distribución AleatoriaRESUMEN
In this paper, we study targeted immunization in a multilayer Boolean network model for genetic regulatory networks. Given a specific set of nodes immune to perturbations, we find that the stability of a multilayer Boolean network is determined by the largest eigenvalue of the weighted non-backtracking matrix of corresponding aggregated network. Aimed to minimize this largest eigenvalue, we developed the metric of multilayer collective influence (MCI) to quantify the impact of immunizing individual nodes on the stability of the system. Compared with other competing heuristics, immunizing nodes with high MCI scores can stabilize an unstable multilayer network with higher efficiency on both synthetic and real-world networks. Moreover, despite that coupling nodes can exert direct influence across multiple layers, they are found to exhibit less importance as measured by the MCI score. Our work reveals the mechanism of maintaining the stability of multilayer Boolean networks and provides an efficient targeted immunization strategy, which can be potentially applied to the location of pathogenesis of diseases and the development of targeted therapy.
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Inmunización , Algoritmos , Simulación por Computador , Redes Reguladoras de Genes/inmunología , Humanos , Modelos GenéticosRESUMEN
Serine 727 (Ser727) phosphorylation of STAT3 plays a role in the regulation of mitochondrial respiration. This study aimed to test if zinc could regulate mitochondrial respiration through phosphorylation of STAT3 at Ser727 in the setting of ischemia/reperfusion in the heart. Under normoxic conditions, treatment of isolated rat hearts with ZnCl2 increased cytosolic STAT3 phosphorylation at Ser727 followed by phospho-STAT3 translocation to mitochondria. In isolated rat hearts subjected to 30â¯min regional ischemia followed by 20â¯min of reperfusion, ZnCl2 given 5â¯min before the onset of reperfusion also increased mitochondrial phospho-STAT3. ZnCl2 enhanced ERK phosphorylation and PD98059 reversed the effect of ZnCl2 on STAT3 phosphorylation. ZnCl2 improved the mitochondrial oxidative phosphorylation at reperfusion. This effect was abolished by STAT3S727A, a mutant in which Ser727 is replaced with alanine, in H9c2 cells subjected to hypoxia/reoxygenation. In addition, ZnCl2 increased the mRNA level of the complex I subunit ND6, which was also reversed by STAT3S727A. Moreover, ZnCl2 attenuated mitochondrial ROS generation and dissipation of mitochondrial membrane potential (ΔΨm) at reoxygenation through Ser727 phosphorylation. Finally, ZnCl2 suppression of succinate dehydrogenase (SDH) activity upon the onset of reperfusion was nullified by the Ser727 mutation. In conclusion, zinc improves cardiac oxidative phosphorylation and inhibits mitochondrial ROS generation at reperfusion by increasing mitochondrial STAT3 phosphorylation at Ser727 via ERK. The preservation of ND6 mtDNA and the inhibition of SDH activity may account for the role of STAT3 in the beneficial action of zinc on the mitochondrial oxidative phosphorylation and ROS generation at reperfusion.
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Mitocondrias/metabolismo , Daño por Reperfusión Miocárdica/metabolismo , Fosfoserina/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Zinc/farmacología , Animales , Línea Celular , Respiración de la Célula/efectos de los fármacos , Etilenodiaminas/farmacología , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Masculino , Mitocondrias/efectos de los fármacos , Daño por Reperfusión Miocárdica/patología , Fosforilación/efectos de los fármacos , Ratas Wistar , Factor de Transcripción STAT3/metabolismo , Succinato Deshidrogenasa/metabolismoRESUMEN
Background: Chronic kidney disease (CKD) is a common complication of diabetes, and requires long-term medical care. However, besides the blood glucose level, no reliable method is currently available to link the quality of care and the development of CKD. We therefore developed a long-term quality-of-care score for predicting the occurrence of CKD in patients with type 2 diabetes. Methods: In this retrospective cohort study, using Taiwan's Longitudinal Cohort of Diabetes Patients Database and the medical records in a medical center, we identified incident patients with type 2 diabetes during 1999-2003 and followed them until 2011. A quality-of-care score (from 0 to 8) was calculated according to process indicators (frequencies of HbA1c and lipid profile testing and urine, foot, and retinal examinations), intermediate outcome indicators (low-density lipoprotein, blood pressure, and HbA1c) and comorbidity of hypertension. We used Cox regression models to evaluate the association between the score and the incidence of CKD. Results: Of the 4754 patients enrolled, 1407 developed CKD after a mean follow-up of 9.06 years. Compared with the risk of developing a CKD event in patients with scores ≤2, the risk was 69% lower in those with quality-of-care scores ≥5 (hazard ratio [HR] 0.31; 95% confidence interval [CI] 0.25-0.40) and 33% lower in those with scores between 3 and 4 (HR 0.67; 95% CI 0.59-0.77). Conclusions: Good quality of care can reduce the risk of CKD in patients with type 2 diabetes. The score developed in this study had a significant association with the risk of CKD and thus can be applied to guide the care for these patients.
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Diabetes Mellitus Tipo 2/complicaciones , Diabetes Mellitus Tipo 2/terapia , Calidad de la Atención de Salud/normas , Insuficiencia Renal Crónica/epidemiología , Adulto , Anciano , Presión Sanguínea/fisiología , Estudios de Cohortes , Comorbilidad , Femenino , Hemoglobina Glucada/análisis , Humanos , Hipertensión/complicaciones , Incidencia , Lípidos/análisis , Lipoproteínas LDL/análisis , Masculino , Persona de Mediana Edad , Modelos de Riesgos Proporcionales , Calidad de la Atención de Salud/estadística & datos numéricos , Insuficiencia Renal Crónica/prevención & control , Estudios Retrospectivos , Taiwán/epidemiologíaRESUMEN
BACKGROUND: Although it is well known that remifentanil (Rem) elicits cardiac protection against ischemia/reperfusion (I/R) injury, the underlying mechanism remains unclear. This study tested if Rem can protect the heart from I/R injury by inhibiting endoplasmic reticulum (ER) stress through the maintenance of zinc (Zn) homeostasis. METHODS: Isolated rat hearts were subjected to 30 minutes of regional ischemia followed by 2 hours of reperfusion. Rem was given by 3 consecutive 5-minute infusions, and each infusion was followed by a 5-minute drug-free perfusion before ischemia. Total Zn concentrations in cardiac tissue, cardiac function, infarct size, and apoptosis were assessed. H9c2 cells were subjected to 6 hours of hypoxia and 2 hours of reoxygenation (hypoxia/reoxygenation [H/R]), and Rem was given for 30 minutes before hypoxia. Metal-responsive transcription factor 1 (MTF1) overexpression plasmids were transfected into H9c2 cells 48 hours before hypoxia. Intracellular Zn level, cell viability, and mitochondrial injury parameters were evaluated. A Zn chelator N,N,N',N'-tetrakis-(2-pyridylmethyl) ethylenediamine (TPEN) or an ER stress activator thapsigargin was administrated during in vitro and ex vivo studies. The regulatory molecules related to Zn homeostasis and ER stress in cardiac tissue, and cardiomyocytes were analyzed by Western blotting. RESULTS: Rem caused significant reversion of Zn loss from the heart (Rem + I/R versus I/R, 9.43 ± 0.55 vs 7.53 ± 1.18; P < .05) by suppressing the expression of MTF1 and Zn transporter 1 (ZnT1). The inhibited expression of ER stress markers after Rem preconditioning was abolished by TPEN. Rem preconditioning improved the cardiac function accompanied by the reduction of infarct size (Rem + I/R versus I/R, 21% ± 4% vs 40% ± 6%; P < .05). The protective effects of Rem could be reserved by TPEN and thapsigargin. Similar effects were observed in H9c2 cells exposed to H/R. In addition, MTF1 overexpression blocked the inhibitory effects of Rem on ZnT1 expression and ER stress at reoxygenation. Rem attenuated the collapse of mitochondrial membrane potential (ΔΨm) and the generation of mitochondrial reactive oxygen species by inhibiting ER stress via cardiac Zn restoration (Rem + H/R versus H/R, 79.57% ± 10.62% vs 58.27% ± 4.32%; P < .05). CONCLUSIONS: Rem maintains Zn homeostasis at reperfusion by inhibiting MTF1 and ZnT1 expression, leading to the attenuation of ER stress and cardiac injury. Our findings provide a promising therapeutic approach for managing acute myocardial I/R injury.
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Estrés del Retículo Endoplásmico/efectos de los fármacos , Infarto del Miocardio/prevención & control , Daño por Reperfusión Miocárdica/prevención & control , Miocitos Cardíacos/efectos de los fármacos , Remifentanilo/farmacología , Zinc/metabolismo , Animales , Apoptosis/efectos de los fármacos , Proteínas de Transporte de Catión/genética , Proteínas de Transporte de Catión/metabolismo , Línea Celular , Citoprotección , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Modelos Animales de Enfermedad , Homeostasis , Preparación de Corazón Aislado , Masculino , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Mitocondrias Cardíacas/efectos de los fármacos , Mitocondrias Cardíacas/metabolismo , Mitocondrias Cardíacas/patología , Infarto del Miocardio/metabolismo , Infarto del Miocardio/patología , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/patología , Daño por Reperfusión Miocárdica/fisiopatología , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/patología , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Función Ventricular Izquierda/efectos de los fármacos , Factor de Transcripción MTF-1RESUMEN
A novel organic monolith was successfully fabricated by a one-pot thiol-ene click reaction of triallyl isocyanurate with pentaerythritol tetrakis-(2-mercaptoacetate) and mercaptopropionic acid in the presence of porogens. We investigated the effects of the ratio of monomer and cross-linking agent, the type and ratio of porogen, and click reaction temperature on the permeability and morphology of the prepared poly triallyl isocyanurate-co-pentaerythritol tetrakis (2-mercaptoacetate) monoliths. The monolith was also characterized by scanning electron microscopy and Fourier transform infrared spectroscopy. The results indicated that the monoliths had continuous porous framework, good permeability, and high mechanical stability. A series of analytes with different properties such as alkylbenzenes, polycyclic aromatic hydrocarbons, anilines, and phenols were used to evaluate the electrochromatographic performance of the prepared monoliths in pressurized capillary electrochromatography. The prepared polymer monolith showed typical reversed-phase electrochromatographic behavior for hydrophobic substances. Moreover, the prepared monolith showed a mix of reversed-phase and cation exchange interaction modes for basic aniline compounds. The minimum plate height of the monolith was 8.76 µm (132 100 plates/m) for propylbenzene. These results demonstrated that one-pot thiol-ene click chemistry can provide a simple and reliable method for the preparation of organic monoliths.
RESUMEN
Voltage-gated sodium channels, which are involved in pain pathways, have emerged as major targets for therapeutic intervention in pain disorders. Nav1.7, the tetrodotoxin-sensitive voltage-gated sodium channel isoform encoded by SCN9A and predominantly expressed in pain-sensing neurons in the dorsal root ganglion, plays a crucial role in nociception. MicroRNAs are highly conserved, small non-coding RNAs. Through binding to the 3' untranslated region of their target mRNAs, microRNAs induce the cleavage and/or inhibition of protein translation. Based on bioinformatics analysis using TargetScan software, we determined that miR-30b directly targets SCN9A To investigate the roles of Nav1.7 and miR-30b in neuropathic pain, we examined changes in the expression of Nav1.7 in the dorsal root ganglion by miR-30b over-expression or knockdown in rats with spared nerve injury. Our results demonstrated that the expression of miR-30b and Nav1.7 was down-regulated and up-regulated, respectively, in the dorsal root ganglion of spared nerve injury rats. MiR-30b over-expression in spared nerve injury rats inhibited SCN9A transcription, resulting in pain relief. In addition, miR-30b knockdown significantly increased hypersensitivity to pain in naive rats. We also observed that miR-30b decreased Nav1.7 expression in PC12 cells. Taken together, our results suggest that miR-30b plays an important role in neuropathic pain by regulating Nav1.7 expression. Therefore, miR-30b may be a promising target for the treatment of chronic neuropathic pain.