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OBJECTIVE: Long QT syndrome type 7 (Andersen-Tawil syndrome, ATS), which is caused by KCNJ2 gene mutation, often leads to ventricular arrhythmia, periodic paralysis and skeletal malformations. The development, differentiation and electrophysiological maturation of cardiomyocytes (CMs) changes promote the pathophysiology of Long QT syndrome type 7(LQT7). We aimed to specifically reproduce the ATS disease phenotype and study the pathogenic mechanism. METHODS AND RESULTS: We established a cardiac cell model derived from human induced pluripotent stem cells (hiPSCs) to the phenotypes and electrophysiological function, and the establishment of a human myocardial cell model that specifically reproduces the symptoms of ATS provides a reliable platform for exploring the mechanism of this disease or potential drugs. The spontaneous pulsation rate of myocardial cells in the mutation group was significantly lower than that in the repair CRISPR group, the action potential duration was prolonged, and the Kir2.1 current of the inward rectifier potassium ion channel was decreased, which is consistent with the clinical symptoms of ATS patients. Only ZNF528, a chromatin-accessible TF related to pathogenicity, was continuously regulated beginning from the cardiac mesodermal precursor cell stage (day 4), and continued to be expressed at low levels, which was identified by WGCNA method and verified with ATAC-seq data in the mutation group. Subsequently, it indicated that seven pathways were downregulated (all p < 0.05) by used single sample Gene Set Enrichment Analysis to evaluate the overall regulation of potassium-related pathways enriched in the transcriptome and proteome of late mature CMs. Among them, the three pathways (GO: 0008076, GO: 1990573 and GO: 0030007) containing the mutated gene KCNJ2 is involved that are related to the whole process by which a potassium ion enters the cell via the inward rectifier potassium channel to exert its effect were inhibited. The other four pathways are related to regulation of the potassium transmembrane pathway and sodium:potassium exchange ATPase (p < 0.05). ZNF528 small interfering (si)-RNA was applied to hiPSC-derived cardiomyocytes for CRISPR group to explore changes in potassium ion currents and growth and development related target protein levels that affect disease phenotype. Three consistently downregulated proteins (KCNJ2, CTTN and ATP1B1) associated with pathogenicity were verificated through correlation and intersection analysis. CONCLUSION: This study uncovers TFs and target proteins related to electrophysiology and developmental pathogenicity in ATS myocardial cells, obtaining novel targets for potential therapeutic candidate development that does not rely on gene editing.
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Síndrome de Andersen , Células Madre Pluripotentes Inducidas , Humanos , Síndrome de Andersen/diagnóstico , Síndrome de Andersen/genética , Cromatina/metabolismo , Transcriptoma , Mutación/genética , Miocitos Cardíacos/metabolismo , Potasio/metabolismoRESUMEN
OBJECTIVE: To explore the genetic basis for a child with optic atrophy and global developmental delay. METHODS: A child who had presented at the Guangzhou Women and Children's Medical Center in January 2022 was selected as the study subject. Clinical data were collected. Whole exome sequencing (WES) was carried out for the child. Candidate variant was validated by Sanger sequencing and bioinformatic analysis. RESULTS: The child, a nine-month-old female, had manifested dysopia and global developmental delay. Genetic testing revealed that she has harbored a de novo c.425G>C (p.Arg142Pro) variant of the NR2F1 gene, which has been associated with Bosch-Boonstra-Schaaf syndrome. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as pathogenic (PS2+PM1+PM2_Supporting+PM5+PP3+PP4). CONCLUSION: The c.425G>C (p.Arg142Pro) variant of the NR2F1 gene probably underlay the pathogenesis in this child. Above finding has enriched the genotypic and phenotypic spectrum of the NR2F1 gene.
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Atrofia Óptica , Femenino , Humanos , Lactante , Biología Computacional , Factor de Transcripción COUP I/genética , Pruebas Genéticas , Genómica , Genotipo , Atrofia Óptica/genéticaRESUMEN
OBJECTIVE: To explore the genetic basis for a child featuring global developmental disorder with epilepsy. METHODS: A child who had presented at Guangzhou Women and Children's Medical Center in July 2022 was selected as the study subject. Clinical data was collected. Potential variant was detected by whole exome sequencing (WES). Candidate variant was validated by Sanger sequencing and bioinformatic analysis. RESULTS: The child, a three-year-old ethnic Zhuang Chinese girl, had presented with global developmental disorder and epilepsy, for which rehabilitation therapy was ineffective. Genetic testing revealed that she has harbored a homozygous c.821T>C (p.Leu274Pro) missense variant of the PIGW gene, for which both of her parents and sister were heterozygous carriers. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as variant of uncertain significance. CONCLUSION: The homozygous c.821T>C (p.Leu274Pro) variant of the PIGW gene probably underlay the onset of disease in this child. Above finding has enriched the mutational spectrum of the PIGW gene.
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Discapacidades del Desarrollo , Epilepsia , Preescolar , Femenino , Humanos , Biología Computacional , Epilepsia/genética , Pruebas Genéticas , HomocigotoRESUMEN
ATP-binding cassette transporter G1 (ABCG1) is a cellular transmembrane protein that transports oxysterol efflux from cells to high-density lipoprotein (HDL) particles in the plasma. Previous studies have demonstrated that an ABCG1 deficiency exerts an antiatherosclerotic function through the effects of oxysterol accumulation in cells to enhance apoptosis and regulate inflammatory processes. However, whether the deficiency of ABCG1 and the corresponding changes in the efflux of oxysterols could take a series of impacts on the proteomic composition of HDL remains unclear. Here, plasma HDL of ABCG1(-/-) mice and their wild-type controls on a normal chow diet (NCD) or a high-fat diet (HFD) were isolated by ultracentrifugation. The proportion of 7-ketocholesterol and the proteomic composition of samples were comparatively analyzed by LC-MS/MS. In NCD-fed mice, lipid metabolism-related protein (arachidonate 12-lipoxygenase) and antioxidative protein (pantetheinase) exhibited increased accumulation, and inflammatory response protein (alpha-1-antitrypsin) was decreased in accumulation in ABCG1(-/-) mice HDL. In HFD-fed mice, fewer proteins were detected than that of NCD-fed mice. The ABCG1(-/-) mice HDL exhibited increased accumulation of lipid metabolism-related proteins (e.g., carboxylesterase 1C, apolipoprotein (apo)C-4) and decreased accumulation of alpha-1-antitrypsin, as well as significantly reduced proportion of 7-ketocholesterol. Additionally, positive correlations were found between 7-ketocholesterol and some essential proteins on HDL, such as alpha-1-antitrypsin, apoA-4, apoB-100, and serum amyloid A (SAA). These results suggest a detrimental impact of oxysterols on HDL composition, which might affect the antiatherosclerotic properties of HDL.
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Dieta Alta en Grasa , Enfermedades no Transmisibles , Transportador de Casetes de Unión a ATP, Subfamilia G, Miembro 1/genética , Animales , Cromatografía Liquida , Dieta Alta en Grasa/efectos adversos , Lipoproteínas/metabolismo , Ratones , Ratones Noqueados , Proteómica , Espectrometría de Masas en TándemRESUMEN
The present study aimed to explore the pathogenesis of autoimmune myocarditis induced by PD-1 inhibitors and their potential therapeutic targets. Mouse models of autoimmune myocarditis induced by PD-1 inhibitor in mouse models of polymyositis were established. The expression level of PD-1 and regulatory T cells (Tregs), CD4, CD8 + T cells, inflammation, apoptosis and autophagy-related factors, including IL-6, TGF-ß, AMA-M2, Fas/FasL, LC3 and p62 were detected in peripheral blood, muscle or myocardium of mice in each group, using ELISA, RT-PCR, Western Blot and immunofluorescence. In addition, HE and TUNEL staining and ultrastructural scanning were performed on the myocardium of mice in each group. Results showed that the expression level of PD-1 in the two myositis groups was significantly lower than that in the control group, and the level of PD-1 was lower in the myocarditis group than that in the polymyositis group. In the myocardium, TGF-ß, p62, and Tregs proportion showed the same expression level trend as PD-1, while CD8, IL-6, IL-10 and LC3 showed the opposite trend. Levels of Fas/FasL were significantly higher in both myositis groups, but were slightly lower in the myocarditis group, as was AMA-M2. Inflammation, apoptosis, and autophagy were observed in both myositis groups, but were more severe in the myocarditis group. In summary, the decreased expression level of PD-1 leads to decreased Tregs level in the myocardium, aggravated inflammatory response, apoptosis and autophagy, which may be the pathological mechanism of myocarditis induced by PD-1 inhibitors.
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Miocarditis , Miositis , Polimiositis , Animales , Apoptosis , Autofagia , Inhibidores de Puntos de Control Inmunológico , Inflamación/patología , Interleucina-6/uso terapéutico , Ratones , Miocardio/patología , Miositis/tratamiento farmacológico , Miositis/patología , Polimiositis/patología , Receptor de Muerte Celular Programada 1 , Linfocitos T Reguladores/metabolismo , Factor de Crecimiento Transformador betaRESUMEN
BACKGROUND: This study investigated neutrophil activation and neutrophil-derived extracellular traps formation in coronary artery ectasia. METHODS: We enrolled 90 patients who underwent coronary angiography, and included 30 patients with coronary artery ectasia (CAE), 30 patients with obstructive coronary artery disease (CAD) and 30 patients with normal coronary arteries (CON). Intra-neutrophil mean myeloperoxidase index (MPXI) was determined using an automated blood cell counter (ADVIA2120 Hematology System). Serum concentrations of plasma adhesion molecules, cytokines, and neutrophil-derived extracellular traps were quantified. RESULTS: The intra-neutrophil mean myeloperoxidase index was reduced in CAE patients compared to CAD and CON patients (1.02 ± 3.01, 3.22 ± 3.03, 3.52 ± 4.25, respectively; CAE vs CAD, p = 0.016 and CAE vs CON, p = 0.007). Multiple logistic regression analysis showed that MPXI and dsDNA were independent factors that predicted the presence of CAE. CAE patients had higher levels of plasma adhesion molecules (P-selectin glycoprotein ligand-1, E-selectin, L-selectin) and interleukin 1 beta levels. Neutrophil extracellular trap concentrations were significantly higher in the CAE group compared to CAD and CON patients (284.31(258.33-449.91) ng/mL, 225.12(203.34-257.13) ng/mL, and 247.37(231.04-273.01) ng/mL, respectively; CAE vs CAD, p = 0.000 and CAE vs CON, p = 0.001). CONCLUSIONS: Peripheral neutrophils from CAE patients were activated and neutrophil extracellular traps were elevated in the plasma. IL-1ß and soluble adhesion molecules may be the causal factors for neutrophil activation.
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Enfermedad de la Arteria Coronaria/metabolismo , Vasos Coronarios/metabolismo , Trampas Extracelulares/metabolismo , Activación Neutrófila , Neutrófilos/metabolismo , Adulto , Anciano , Estudios de Casos y Controles , Moléculas de Adhesión Celular/sangre , Angiografía Coronaria , Enfermedad de la Arteria Coronaria/diagnóstico por imagen , Enfermedad de la Arteria Coronaria/inmunología , Vasos Coronarios/diagnóstico por imagen , Vasos Coronarios/inmunología , Dilatación Patológica , Trampas Extracelulares/inmunología , Femenino , Humanos , Interleucina-1beta/sangre , Masculino , Persona de Mediana Edad , Neutrófilos/inmunología , Peroxidasa/metabolismo , Estudios ProspectivosRESUMEN
The effects of high sodium intake on the functionality of resistance arteries have been repeatedly studied in vitro, but no study has focused on salt-sensitive hypertension in vivo. We studied the in vivo reactivity of mesenteric small arteries (MSAs) to vasoactive agents in Dahl salt-sensitive (DS) rats with various sodium diets. Twenty-four male DS rats were randomized into 3 groups: LS (0.3% NaCl diet), NS (0.6% NaCl diet), and HS (8% NaCl diet). After a 12-week intervention, the diameter changes of the MSAs after noradrenaline (NA) and acetylcholine (ACh) exposure were detected by a microscope, and changes in blood perfusion through the MSAs were measured by full-field laser perfusion imaging. HS enhanced the constrictive response of the MSAs to NA and attenuated the relaxing response to ACh. Low sodium intake reduced the response of the MSAs to NA and promoted ACh-induced vasodilatation. HS also aggravated NA-induced blood perfusion reduction and impaired ACh-induced hyperperfusion of the MSAs. Pathologically, HS was associated with arteriolar structural damage and fibrosis of the MSAs. We conclude that sodium intake affects the responsiveness of the MSAs to vasoactive agents in DS rats and might play important roles in modulating blood pressure in hypertensive individuals.
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Hipertensión/fisiopatología , Arterias Mesentéricas/fisiopatología , Cloruro de Sodio Dietético , Vasoconstricción , Vasodilatación , Animales , Velocidad del Flujo Sanguíneo , Dieta Hiposódica , Modelos Animales de Enfermedad , Fibrosis , Hipertensión/etiología , Hipertensión/metabolismo , Hipertensión/patología , Masculino , Arterias Mesentéricas/metabolismo , Arterias Mesentéricas/patología , Óxido Nítrico Sintasa de Tipo III/metabolismo , Peptidil-Dipeptidasa A/metabolismo , Ratas Endogámicas Dahl , Receptor de Angiotensina Tipo 1/metabolismo , Circulación Esplácnica , Remodelación Vascular , Resistencia VascularRESUMEN
BACKGROUND: X-linked agammaglobulinaemia (XLA) is a rare inherited primary immunodeficiency disease characterized by the B cell developmental defect, caused by mutations in the gene coding for Bruton's tyrosine kinase (BTK), which may cause serious recurrent infections. The diagnosis of XLA is sometimes challenging because a few number of patients have higher levels of serum immunoglobulins than expected. In this study, we reported an atypical case with recurrent meningitis, delayed diagnosis with XLA by genetic analysis at the second episode of meningitis at the age of 8 years. CASE REPORT: An 8-year-old Chinese boy presented with fever, dizziness and recurrent vomiting for 3 days. The cerebrospinal fluid (CSF) and magnetic resonance imaging (MRI) results were suggestive of bacterial meningoencephalitis, despite the negative gram staining and cultures of the CSF. The patient was treated with broad-spectrum antibiotics and responded well to the treatment. He had history of another episode of acute pneumococci meningitis 4 years before. The respective level of Immunoglobulin G (IgG), Immunoglobulin A (IgA) and Immunoglobulin M (IgM) was 4.85 g/L, 0.93 g/L and 0.1 g/L at 1st episode, whereas 1.9 g/L, 0.27 g/L and 0 g/L at second episode. The B lymphocytes were 0.21 and 0.06% of peripheral blood lymphocytes at first and second episode respectively. Sequencing of the BTK coding regions showed that the patient had a point mutation in the intron 14, hemizyous c.1349 + 5G > A, while his mother had a heterozygous mutation. It was a splice site mutation predicted to lead to exon skipping and cause a truncated BTK protein. CONCLUSION: Immunity function should be routinely checked in patients with severe intracranial bacterial infection. Absence of B cells even with normal level of serum immunoglobulin suggests the possibility of XLA, although this happens only in rare instances. Mutational analysis of BTK gene is crucial for accurate diagnosis to atypical patients with XLA.
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Agammaglobulinemia/complicaciones , Agammaglobulinemia/diagnóstico , Enfermedades Genéticas Ligadas al Cromosoma X/complicaciones , Enfermedades Genéticas Ligadas al Cromosoma X/diagnóstico , Encefalitis Infecciosa/genética , Agammaglobulinemia Tirosina Quinasa/genética , Agammaglobulinemia/genética , Niño , Análisis Mutacional de ADN , Diagnóstico Tardío , Enfermedades Genéticas Ligadas al Cromosoma X/genética , Humanos , Masculino , MutaciónRESUMEN
BACKGROUND Leukocyte telomere length (LTL) is regarded as a potential marker of biological aging. Oxidative stress plays a major role in the rate of telomeric DNA loss. The aim of this study was to explore whether the LTL was shorter in Chinese patients with premature coronary artery disease (PCAD) than in non-CAD controls and to determine the relationship between oxidative stress and LTL shortening in this population. MATERIAL AND METHODS Patients for coronary angiography were recruited. In total, 128 patients with PCAD and 128 non-CAD controls were enrolled. Samples of circulating leukocytes and plasma were collected. The mean LTL was measured using a polymerase chain reaction-based assay and expressed as the ratio of telomere repeat copies to single-copy gene (SCG) copies (T/S ratio). Reactive oxygen species (ROS) levels and total antioxidant capacity (T-AOC) were determined in plasma. RESULTS Both the T/S ratio (0.88±0.86 vs. 1.10±0.57, P=0.015) and telomere base pairs (4.97±1.37 kb vs. 5.32±0.91 kb, P=0.015) were significantly shorter in the PCAD group than in non-CAD controls. The T-AOC levels of the PCAD group were significantly lower than those of the non-CAD controls (0.482 mM [0.279, 0.603 mM]) vs. 0.778 mM [0.421, 0.924 mM], P=0.000). The ratio of T-AOC to ROS in the PCAD patients was significantly decreased compared to that of the non-CAD controls (0.1026±0. 1587 [Mm*ml/ng] vs. 0.1435±0.1946 [Mm*ml/ng], P=0.013). CONCLUSIONS The results point to a potential link between reduced LTLs in patients with PCAD and early onset of atherosclerosis. The decline in antioxidant capacity may play an important role in accelerating the attrition of telomeres in PCAD patients.
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Enfermedad de la Arteria Coronaria/genética , Estrés Oxidativo/genética , Telómero/fisiología , Adulto , Anciano , Pueblo Asiatico/genética , Aterosclerosis/genética , Aterosclerosis/fisiopatología , Biomarcadores/sangre , China , Enfermedad de la Arteria Coronaria/fisiopatología , Femenino , Humanos , Leucocitos/fisiología , Masculino , Persona de Mediana Edad , Estrés Oxidativo/fisiología , Especies Reactivas de Oxígeno/sangre , Especies Reactivas de Oxígeno/metabolismo , Telómero/genética , Homeostasis del Telómero/genética , Homeostasis del Telómero/fisiologíaRESUMEN
OBJECTIVES: This study sought to investigate the clinical correlates and prognostic roles of urokinase-type plasminogen activator receptor (uPAR) on circulating monocytes in patients with coronary artery disease (CAD). METHODS: 263 angina patients were included in this study. The percentage of uPAR expressing monocytes (PUEM) and the mean fluorescence intensity (MFI) index of uPAR were measured using flow cytometry. Patient follow-up was on average 604 days. Major adverse cardiac events (MACE) were defined as a composite of cardiac death, reinfarction, acute heart failure and hospitalization for revascularization. RESULTS: The PUEM and MFI index levels were significantly more elevated in acute coronary syndrome patients than in stable ones. uPAR expressions on circulating monocytes at admission were correlated to inflammatory biomarkers and myocardial necrosis. Logistic regression analysis revealed that PUEM ≥15% (OR 21.96, 95% CI 7.31-65.98, p < 0.001) and uPAR MFI index ≥3.00 (OR 3.54, 95% CI 1.18-10.59, p = 0.024) were independent determinants of clinical instability in patients with CAD. When followed up, a high PUEM level at admission was an independent prognostic parameter for long-term MACE (HR 3.99, 95% CI 1.31-12.11, p = 0.015). CONCLUSIONS: uPAR expression on circulating monocytes is associated with clinical instability and myocardial necrosis and independently predicts the risk of MACE in patients with CAD.
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Receptores del Activador de Plasminógeno Tipo Uroquinasa/metabolismo , Enfermedad Aguda , Anciano , Angina de Pecho , Biomarcadores/metabolismo , Angiografía Coronaria , Enfermedad de la Arteria Coronaria , Femenino , Insuficiencia Cardíaca/etiología , Hospitalización/estadística & datos numéricos , Humanos , Masculino , Persona de Mediana Edad , Monocitos/metabolismo , Infarto del Miocardio/etiología , Pronóstico , Estudios Prospectivos , RecurrenciaRESUMEN
Dysregulation of the metabolism of the extracellular matrix (ECM) may contribute to coronary artery ectasia (CAE). This study evaluated the turnover of main ECM components and related proteolytic enzymes activities. In this study, thirty patients with CAE, 30 patients with coronary artery disease (CAD) and 30 subjects with normal coronary arteries (Control) were selected. The following circulating ECM metabolism markers were measured: soluble elastin (sElastin), collagen type I cross-linked telopeptides (ICTP), procollagen type I carboxy terminal peptide (PICP), protocollagen III N-terminal propeptide (PIIINP), and procollagen a1(III) C-terminal propeptide (PIIICP). Serum total elastase activity and total matrix metalloproteinase (MMP) activity were also determined. The level of sElastin was higher in the CAE group than in the CAD and Control groups (P1 = 0.009, P2 = 0.000). There was no difference in ICTP (P = 0.168) or PIIICP (P = 0.079) among the three groups. PICP was significantly elevated in CAE (P1 = 0.001, P2 = 0.002). PIIINP was also significantly increased in CAE (P1 = 0.002, P2 = 0.007). Total elastase activity was higher in the CAE group than in the other two groups (P1 = 0.006, P2 = 0.022). Total MMP activity was significantly higher in the CAE group than the Control group (P2 = 0.013) but not higher than the CAD group (P1 = 0.477). In conclusion, within CAE patients the main changes in ECM metabolism were increased degradation of elastin fibres and the transition of collagen from type III to type I. Elastase and MMPs appear to be associated with this kind of ECM turnover.
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Enfermedad de la Arteria Coronaria/sangre , Vasos Coronarios/metabolismo , Matriz Extracelular/metabolismo , Fragmentos de Péptidos/sangre , Anciano , Biomarcadores/sangre , Estudios de Casos y Controles , Colágeno Tipo I/sangre , Colágeno Tipo III/sangre , Enfermedad de la Arteria Coronaria/diagnóstico por imagen , Vasos Coronarios/diagnóstico por imagen , Dilatación Patológica , Elastina/sangre , Femenino , Humanos , Masculino , Metaloproteinasas de la Matriz/metabolismo , Persona de Mediana Edad , Elastasa Pancreática/metabolismo , ProteolisisRESUMEN
OBJECTIVE: Proteolytic enzymes might contribute to coronary artery ectasia (CAE) through the destruction of extracellular matrix (ECM) vessel components. This study aimed to find out if peripheral blood mononuclear cells (PBMCs) served as a source of those proteinases and their regulators. METHOD: In this study, transcriptional expression profiles of the main proteinases and their regulators were detected in the PBMCs of CAE patients as follows: (1) matrix metalloproteinases (MMP) 1, 2, 3, 8, 9, 10, 12 and 13; (2) the serine proteinases elastase: cathepsin G and proteinases 3; (3) the cysteine proteinases: cathepsin L and cathepsin S; (4) the main endogenous inhibitors for the above proteinases: tissue inhibitors of metalloproteinase (TIMP) 1 and 2, α1-antitrypsin (α1-PI) and α2-macroglobulin (A2M); (5) twelve cytokines that could regulate the above proteinases. RESULT: The characteristic changes in CAE were: (1) MMP1 and MMP9 increased while the serine and cysteine families did not change; (2) the four proteinase inhibitors did not change in the CAE group; (3) among the 12 cytokines, interleukin-1 alpha (IL1A), platelet-derived growth factor (PDGF), interferon gamma (IFNγ) and growth differentiation factor 15 (GDF15) were elevated. Partial correlation analysis showed that MMP1 significantly correlated with IL1A and with IFNγ, and MMP9 correlated with IFNγ and with GDF 15. CONCLUSION: PBMCs might participate in the pathological process of CAE by the increased expression of MMP1, MMP9, IL1A, IFNγ and GDF15.
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Enfermedad de la Arteria Coronaria , Vasos Coronarios/patología , Leucocitos Mononucleares/enzimología , Anciano , Angiografía Coronaria/métodos , Enfermedad de la Arteria Coronaria/sangre , Enfermedad de la Arteria Coronaria/diagnóstico , Enfermedad de la Arteria Coronaria/enzimología , Dilatación Patológica/sangre , Dilatación Patológica/diagnóstico , Dilatación Patológica/enzimología , Femenino , Factor 15 de Diferenciación de Crecimiento/metabolismo , Humanos , Interferón gamma/metabolismo , Interleucina-1alfa/metabolismo , Masculino , Metaloproteinasa 1 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Persona de Mediana Edad , TranscriptomaRESUMEN
We investigated ATP-binding cassette transporters A1/G1 expression and function in mediating cholesterol efflux by examining the macrophages of cigarette-smoking patients with coronary artery disease (CAD) before and after smoking abstinence. Peripheral blood monocyte cells were collected from nonsmokers (n = 17), non-CAD (NCAD) smokers (n = 35), and CAD smokers (n = 32) before and after 3 months of smoking cessation. We found that the ABCA1 expression level was lower in macrophages from NCAD and CAD smokers than from nonsmokers at baseline. The ABCA1 function of mediating cholesterol efflux was reduced in NCAD and CAD smokers as compared with nonsmokers. After 3 months of smoking cessation, ABCA1 expression and function were improved in CAD smokers. However, ABCG1 expression and function did not change after smoking cessation. Furthermore, ABCA1 expression was inhibited by tar in human acute monocytic leukemia cell line THP-1-derived macrophages through the inhibition of liver X receptors. Nicotine and carbon monoxide did not inhibit ABCA1 expression. Our results indicate that chronic cigarette smoking impaired ABCA1-mediated cholesterol efflux in macrophages and that tobacco abstinence reversed the function and expression of ABCA1, especially in CAD patients. It was tobacco tar, rather than nicotine or carbon monoxide, that played a major role in the tobacco-induced disturbance of cellular cholesterol homeostasis.
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Colesterol/sangre , Enfermedad de la Arteria Coronaria/sangre , Macrófagos/metabolismo , Cese del Hábito de Fumar , Fumar/sangre , Transportador 1 de Casete de Unión a ATP/biosíntesis , Adulto , Anciano , Anciano de 80 o más Años , Línea Celular Tumoral , Enfermedad de la Arteria Coronaria/patología , Femenino , Regulación de la Expresión Génica , Humanos , Macrófagos/patología , Masculino , Persona de Mediana Edad , Fumar/patología , Factores de TiempoRESUMEN
BACKGROUND: Obstructive sleep apnea-hypopnea syndrome (OSAHS) is associated with premature atherosclerosis. However, the associated mechanism remains unknown. This study investigates the expression of adenosine triphosphate (ATP)-binding cassette transporter protein A1 (ABCA1) and cellular cholesterol efflux in cultured macrophages from OSAHS patients. METHODS: Of the 18 subjects enrolled in this study, six subjects with apnea-hypopnea index (AHI) <5 were placed into the control group, and 12 subjects with AHI ≥5 were placed into the OSAHS group. Peripheral blood mononuclear cells (PBMCs) from each subject were isolated, purified, cultured, and differentiated into macrophages in vitro. ABCA1 mRNA and protein expression were evaluated by reverse transcription PCR and Western Blot, respectively. Both ABCA1-mediated and autologous serum induced cholesterol efflux were measured by isotopic cholesterol efflux assays. RESULTS: The levels of AHI and high sensitivity C-reactive protein (hsCRP) were significantly higher in the OSAHS group than in the control group. ABCA1 mRNA and protein expressions in PBMCs-derived macrophages were significantly reduced in patients with OSAHS compared to that in controls (p < 0.05). Both ABCA1-mediated and autologous serum-induced cholesterol efflux were significantly lower in the OSAHS group than that in the control group (p = 0.033 and p = 0.01, respectively). Pearson's correlation analysis revealed a negative correlation between AHI and the mRNA (r = -0.7726, p = 0.0007) and protein (r = -0.8112, p = 0.0044) expression of ABCA1, a positive correlation between ABCA1-mediated cholesterol efflux and the minimum oxygen saturation (r = 0.7954, p < 0.0001), and a negative correlation between AHI and autologous serum induced cholesterol efflux (r = -0.7756, p = 0.0002). CONCLUSION: ABCA1 expression and cellular cholesterol efflux in macrophages were significantly decreased in OSAHS patients, which closely correlated with the severity of disease. Our findings provide meaningful insights into the mechanism of atherogenesis in OSAHS patients.
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Transportador 1 de Casete de Unión a ATP/genética , Aterosclerosis/genética , Aterosclerosis/fisiopatología , Colesterol/metabolismo , Macrófagos/fisiología , Apnea Obstructiva del Sueño/genética , Apnea Obstructiva del Sueño/fisiopatología , Adulto , Estudios de Casos y Controles , Células Cultivadas , Femenino , Regulación de la Expresión Génica/genética , Humanos , Técnicas In Vitro , Masculino , Persona de Mediana Edad , ARN Mensajero/genética , Valores de Referencia , Estadística como AsuntoRESUMEN
OBJECTIVE: To observe the effects of coroanry artery ectasia (CAE) patients' pooled serum on the main proteinases and extracellular matrix (ECM) synthesis and explore whether the growth differentiation factor 15(GDF 15) can regulate the characteristic changes induced by CAE patients' pooled serum. METHODS: Serum samples were collected from 32 CAE patients, 30 patients with coronary heart disease (CHD), and 31 subjects with normal coronary arteries (CON) and then mixed in the same volumes by groups. Then human umbilical vein smooth muscle cells were cultured with the media containing 25% pooled serum. After having been disposed, proteinase system and ECM synthesis system were detected in the cell and culture media samples. GDF15 or GDF15 antibodies was added into the 25% pooled serum in each group to observe if GDF 15 could impact the characteristic changes induced by CAE patients' pooled serum. RESULTS: The expression of matrix metalloproteinases (MMP) 1 mRNA in CAE group was significantly higher than CON group (P=0.002) and CHD group (P=0.000), the secretory MMP1 protein and total MMPs activity in culture media were also upregulated in CAE group (both P<0.01). After adding GDF 15 into the culture media (GDF15+CAE group), the MMP1 mRNA ,secretory MMP1 protein, and total MMPs activity were significantly lower than CAE group (all P<0.01), while in the GDF15 antibody+CAE group, the MMP1 mRNA and total MMPs activities were significantly higher than in GDF15+CAE group (both P<0.01), but the secretory MMP1 protein was not different from GDF 15+CAE group (P>0.05). CONCLUSION: The vascular smooth muscle cells may participate in the CAE process mainly by regulating MMPs system but not the elastase 2 or ECM synthesis system, and GDF15 may be an compensatory factor to prohibit the over-destruction of coronary ECM induced by MMPs.
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Enfermedad de la Arteria Coronaria , Biomarcadores/sangre , Dilatación Patológica , Factor 15 de Diferenciación de Crecimiento , Humanos , Metaloproteinasa 1 de la MatrizRESUMEN
OBJECTIVE: To investgate the effects of rapamycin(RPM)and RPM-loaded poly(lactic-co-glycolic)acid(PLGA)nanoparticles(NPs)on the apoptosis of human umbilical arterial vascular smooth muscle cells(HUASMCs)in vitro and expression of bcl-2 and p27(kip1) protein. METHODS: HUASMCs were cultured in vitro and divided to RPM and RPM-PLGA-NPs groups treated at 3 different concentration by 12 and 24 hours,with M231-smooth muscle growth supplements medium and null-PLGA-NPs treated groups as controlled. The apoptosis of HUASMCs was determined by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling staining and flow cytometry. The expressions of bcl-2 and p27(kip1) were detected by streptacidin/peroxidase immunohistochemical method. The effect on cellular proliferation was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromidecolorimetry. RESULTS: The proliferation of HUASMCs was inhibited by RPM and RPM-PLGA-NPs in a dose-dependent manner. DNA electrophoresis showed DNA ladder in RPM and RPM-PLGA-NPs groups and classical scalar strips in control groups. The apoptotic indexes of RPM 100 ng/ml group and RPM-PLGA-NPs 500 ng/ml group detected by flow cytometry were(45.45<2.36)% and(35.04<5.64)%,respectively,which were significantly higher than that of M231-smooth muscle growth supplements control group [(2.60<0.95)%,all P<0.01]. The apoptotic indexes of groups incubated with RPM and RPM-PLGA-NPs for 24 hours were significantly higher than those of groups which incubated for 12 hours(P<0.05,P<0.01). The positive expression indexes(PEI)of p27(kip1) and bcl-2 protein were higher in RPM and RPM-PLGA-NPs groups than that of control groups. The Spearman's rank correlation coefficient test showed that there was no significant correlation between the PEI of p27(kip1) and the apoptotic indexes in the RPM group and RPM-PLGA-NPs group(P>0.05). CONCLUSIONS: Rapamycin-loaded PLGA nanoparticles and rapamycin have similar effects in inhibiting proliferation and inducing apoptosis;meanwhile,they upregulate the expression of p27(kip1) protein without downregulating the expression of bcl-2 protein in HUASMCs in vitro. RPM-PLGA-NPs has more potent pro-apoptotic effect than equivalent dose of RPM but is not linearly correlated with the p27(kip1) expression level.
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Apoptosis , Músculo Liso Vascular , Proliferación Celular , Células Cultivadas , Inhibidor p27 de las Quinasas Dependientes de la Ciclina , Humanos , Etiquetado Corte-Fin in Situ , Ácido Láctico , Miocitos del Músculo Liso , Nanopartículas , Ácido Poliglicólico , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Sirolimus , Arterias UmbilicalesRESUMEN
PURPOSE: Glucose-regulated protein 78 (GRP78) is a chaperone protein in the endoplasmic reticulum (ER). Previous studies have suggested that statins favorably affect ER stress by upregulating GRP78. This study was designed to investigate whether the anti-atherosclerotic effect of atorvastatin is modulated by a GRP78-involved pathway. METHODS: Hamsters were made diabetic and randomly divided into a diabetic control group (DMC), a diabetic group with low-dose atorvastatin (DML, 2.5 mg/kg/day), and a diabetic group with high-dose atorvastatin (DMH, 5 mg/kg/day). Pathological examinations of the aortic root were performed, and the level of GRP78 and CD68 expression in the aortic root was detected by immunohistochemistry and RT-PCR analysis. In vitro THP-1 macrophages were treated with glucose and atorvastatin, and their GRP78 and CD68 protein expression levels were measured by Western blot. Next, with and without co-incubation with the GRP78 inhibitor, deoxynivalenol (DON), CD68 protein expression was again analyzed. RESULTS: We found that in vivo atorvastatin prominently limited the area of macrophage infiltration in the subendothelial spaces of the aortic root in the DML and DMH groups, and significantly inhibited CD68 expression (DML or DMH vs. DMC, all p < 0.001) and increased GRP78 expression (DML or DMH vs. DMC, p < 0.05 ~ 0.001). In vitro Western blot results showed that atorvastatin decreased CD68 and increased GRP78 protein expression in glucose-treated THP-1 macrophages, and the suppressing effect of atorvastatin on CD68 expression was almost abolished by co-incubation with the GRP78 inhibitor. CONCLUSIONS: Our results clearly showed that atorvastatin inhibited CD68 expression through GRP78 regulation, and that GRP78 could exert a protective effect in the early stages of atherosclerosis beyond being a chaperone protein, providing a new perspective into the anti-atherosclerosis mechanism of atorvastatin.
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Antígenos CD/metabolismo , Antígenos de Diferenciación Mielomonocítica/metabolismo , Aorta/efectos de los fármacos , Proteínas de Choque Térmico/metabolismo , Ácidos Heptanoicos/farmacología , Pirroles/farmacología , Transducción de Señal/efectos de los fármacos , Animales , Anticolesterolemiantes/farmacología , Aorta/metabolismo , Aterosclerosis/tratamiento farmacológico , Aterosclerosis/metabolismo , Atorvastatina , Chaperón BiP del Retículo Endoplásmico , Glucosa/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Masculino , Mesocricetus , Tricotecenos/farmacologíaRESUMEN
OBJECTIVE: To evaluation the prevalence of hypertension, diabetes, dyslipidemia in systemic lupus erythematosus (SLE) patients, and investigate the factors that affecting lipid levels in SLE patients. METHODS: A total of 540 adult SLE patients hospitalized in Peking Union Medical College Hospital from March 2010 to March 2013 were retrospectively included (SLE group), and 1 080 gender and age matched (1: 2) healthy controls were selected from our medical examination center (control group). The prevalence rate of hypertension, diabetes, dyslipidemia and the levels of serum lipid were compared between the two groups, the factors affecting lipid levels in SLE patients were also analyzed. RESULTS: The percentage of hypertension, diabetes, dyslipidemia, elevated total cholesterol (TC), elevated triglyceride (TG), decreased high density lipoprotein cholesterol (HDL-C) and elevated low density lipoprotein cholesterol (LDL-C) in SLE patients were significantly higher than those in healthy controls (all P < 0.01) . Compared with the control group, SLE patients had significantly higher TC, TG, LDL-C levels and significantly lower HDL-C levels (all P < 0.01) . Multifactor regression analysis showed that TC and LDL-C levels were positively correlated with lupus nephritis (ß = 0.695,0.437), corticosteroids therapy (ß = 1.195, 0.715), complement C4 levels (ß = 4.817, 3.382) and 24 hours urine protein content (ß = 0.112, 0.078) (all P < 0.01) , but negatively correlated with serum albumin (Alb) (ß = -0.107, -0.077) and high sensitive C reactive protein (hsCRP) levels (ß = -0.021, -0.014) (all P < 0.01). TG levels were positively correlated with lupus nephritis (ß = 0.359) and 24 hours urine protein content (ß = 0.045) (both P < 0.05), negatively correlated with male gender (ß = -0.605), age (ß = -0.014) and Alb levels (ß = -0.053) (P < 0.01 or 0.05). HDL-C levels were positively correlated with age (ß = 0.007), lupus nephritis (ß = 0.188), corticosteroids therapy (ß = 0.342), consecutive 30 days cumulative corticosteroids dose before serum lipid were measured (ß<0.001), and complement C3 levels(ß = 0.351) (all P < 0.01) , negatively correlated with hsCRP levels (ß = -0.005, P < 0.01). Serum lipid levels did not correlate with disease duration, disease activity, corticosteroids therapy time, corticosteroids daily dose before serum lipid measurement, serum creatinine levels and erythrocyte sedimentation rate (ESR) (all P > 0.05). CONCLUSION: The prevalence rate of hypertension, diabetes and dyslipidemia in SLE hospitalized patients are significantly higher compared to normal controls and lipid levels of SLE patients are related to various SLE disease factors.
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Enfermedades Cardiovasculares/complicaciones , Lupus Eritematoso Sistémico/complicaciones , Adulto , Enfermedades Cardiovasculares/epidemiología , Estudios de Casos y Controles , Colesterol , HDL-Colesterol , LDL-Colesterol , Dislipidemias , Femenino , Humanos , Hipertensión , Masculino , Prevalencia , Análisis de Regresión , Factores de Riesgo , TriglicéridosRESUMEN
BACKGROUND: The pathogenesis of chronic thromboembolic pulmonary hypertension (CTEPH) is multifactorial and growing evidence has indicated that hematological disorders are involved. Clonal hematopoiesis of indeterminate potential (CHIP) has recently been associated with an increased risk of both hematological malignancies and cardiovascular diseases. However, the prevalence and clinical relevance of CHIP in patients with CTEPH remains unclear. METHODS: Using stepwise calling on next-generation sequencing data from 499 patients with CTEPH referred to 3 centers between October 2006 and December 2021, CHIP mutations were identified. We associated CHIP with all-cause mortality in patients with CTEPH. To provide insights into potential mechanisms, the associations between CHIP and inflammatory markers were also determined. RESULTS: In total, 47 (9.4%) patients with CTEPH carried at least 1 CHIP mutation at a variant allele frequency of ≥2%. The most common mutations were in DNMT3A, TET2, RUNX1, and ASXL1. During follow-up (mean, 55 months), deaths occurred in 22 (46.8%) and 104 (23.0%) patients in the CHIP and non-CHIP groups, respectively (P<0.001, log-rank test). The association of CHIP with mortality remained robust in the fully adjusted model (hazard ratio, 2.190 [95% CI, 1.257-3.816]; P=0.006). Moreover, patients with CHIP mutations showed higher circulating interleukin-1ß and interleukin-6 and lower interleukin-4 and IgG galactosylation levels. CONCLUSIONS: This is the first study to show that CHIP mutations occurred in 9.4% of patients with CTEPH are associated with a severe inflammatory state and confer a poorer prognosis in long-term follow-up.
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Enfermedades Cardiovasculares , Hipertensión Pulmonar , Humanos , Hematopoyesis Clonal , Hipertensión Pulmonar/etiología , Hipertensión Pulmonar/genética , Hematopoyesis/genética , Enfermedades Cardiovasculares/genética , MutaciónRESUMEN
Recent studies showed that statins reduce ATP-binding cassette transporter A1 expression and ATP-binding cassette transporter A1-mediated cholesterol efflux in macrophages, whereas the effect of statins on ATP-binding cassette transporter G1 (ABCG1), another important lipid transporter, is still unclear. Here, we investigated the expression and functionality of ABCG1 on statins in THP-1-derived macrophages and human peripheral blood mononuclear cells. Simvastatin and atorvastatin were used in this study. Treatment with statins significantly decreased ABCG1-mediated cholesterol efflux in human macrophages (from 33.8% ± 2.8% of control to 22.9% ± 1.7% of 10 µM simvastatin or to 23.3% ± 3.3% of 10 µM atorvastatin; P < 0.01, n = 4), whereas the protein expression of ABCG1 remained unaltered on statins. Further analysis revealed that 2 major ABCG1 isoforms responded to statins differently. The expression of ABCG1-S, which exhibited higher activity in cholesterol efflux than ABCG1-L, was significantly decreased on statins compared with increased expression of ABCG1-L. The results suggested that the proportion change of ABCG1 isoform expressions could contribute to reduced ABCG1 functionality under treatment with statins. The effects of statins on ABCG1 isoform expression and functionality were reversed by low-dose liver X receptor agonist, TO-901317, indicating that statins' downregulation of ABCG1 functionality was likely through liver X receptor-dependent pathway. In conclusion, simvastatin and atorvastatin decreased ABCG1-mediated cholesterol efflux in human macrophages without alteration of total ABCG1 protein level. The proportion change of ABCG1 isoforms expressions may be involved in the down-regulation of ABCG1 functionality by statins, which provided a novel mechanism for the regulation of ABCG1 activity.