RESUMEN
Vascular calcification is a pathological process commonly associated with atherosclerosis, chronic kidney disease, and diabetes. Paraspeckle protein NONO is a multifunctional RNA/DNA binding protein involved in many nuclear biological processes but its role in vascular calcification remains unclear. Here, we observed that NONO expression was decreased in calcified arteries of mice and patients with CKD. We generated smooth muscle-specific NONO-knockout mice and established three different mouse models of vascular calcification by means of 5/6 nephrectomy, adenine diet to induce chronic kidney failure, or vitamin D injection. The knockout mice were more susceptible to the development of vascular calcification relative to control mice, as verified by an increased calcification severity and calcium deposition. Likewise, aortic rings from knockout mice showed more significant vascular calcification than those from control mice ex vivo. In vitro, NONO deficiency aggravated high phosphate-induced vascular smooth muscle cell osteogenic differentiation and apoptosis, whereas NONO overexpression had a protective effect. Mechanistically, we demonstrated that the regulation of vascular calcification by NONO was mediated by bone morphogenetic protein 2 (BMP2). NONO directly bound to the BMP2 promoter using its C-terminal region, exerting an inhibitory effect on the transcription of BMP2. Thus, our study reveals that NONO is a novel negative regulator of vascular calcification, which inhibits osteogenic differentiation of vascular smooth muscle cell and vascular calcification via negatively regulating BMP2 transcription. Hence, NONO may provide a promising target for the prevention and treatment of vascular calcification.
Asunto(s)
Proteína Morfogenética Ósea 2 , Modelos Animales de Enfermedad , Ratones Noqueados , Músculo Liso Vascular , Miocitos del Músculo Liso , Osteogénesis , Insuficiencia Renal Crónica , Transcripción Genética , Calcificación Vascular , Animales , Humanos , Masculino , Ratones , Enfermedades de la Aorta/genética , Enfermedades de la Aorta/prevención & control , Enfermedades de la Aorta/patología , Enfermedades de la Aorta/metabolismo , Apoptosis/efectos de los fármacos , Proteína Morfogenética Ósea 2/metabolismo , Proteína Morfogenética Ósea 2/genética , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Proteínas de Unión al ADN/metabolismo , Proteínas de Unión al ADN/genética , Ratones Endogámicos C57BL , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patología , Miocitos del Músculo Liso/metabolismo , Miocitos del Músculo Liso/patología , Miocitos del Músculo Liso/efectos de los fármacos , Osteogénesis/efectos de los fármacos , Regiones Promotoras Genéticas , Insuficiencia Renal Crónica/patología , Insuficiencia Renal Crónica/metabolismo , Insuficiencia Renal Crónica/genética , Insuficiencia Renal Crónica/prevención & control , Proteínas de Unión al ARN/metabolismo , Proteínas de Unión al ARN/genética , Calcificación Vascular/patología , Calcificación Vascular/prevención & control , Calcificación Vascular/metabolismo , Calcificación Vascular/genética , Calcificación Vascular/etiologíaRESUMEN
Technology at the single-cell level has advanced dramatically in characterizing molecular heterogeneity. These technologies have enabled cell subtype diversity to be seen in all tissues, including atherosclerotic plaques. Critical in atherosclerosis pathogenesis and progression are macrophages. Previous studies have only determined macrophage phenotypes within the plaque, mainly by bulk analysis. However, recent progress in single-cell technologies now enables the comprehensive mapping of macrophage subsets and phenotypes present in plaques. In this review, we have updated and discussed the definition and classification of macrophage subsets in mice and humans using single-cell RNA sequencing. We summarized the different classification methods and perspectives: traditional classification with an updated scoring system, inflammatory macrophages, foamy macrophages, and atherosclerotic-resident macrophages. In addition, some special types of macrophages were identified by specific markers, including IFN-inducible and cavity macrophages. Furthermore, we discussed macrophage subset-specific markers and their functions. In the future, these novel insights into the characteristics and phenotypes of these macrophage subsets within atherosclerotic plaques can provide additional therapeutic targets for cardiovascular diseases.
Asunto(s)
Aterosclerosis , Placa Aterosclerótica , Humanos , Ratones , Animales , Placa Aterosclerótica/metabolismo , Aterosclerosis/metabolismo , Macrófagos/metabolismo , Fenotipo , Análisis de Secuencia de ARN/métodosRESUMEN
RATIONALE: The NLRP3 (NLR [NOD-like receptor] family, pyrin domain containing 3) inflammasome is an important driver of atherosclerosis. Our previous study shows that chaperone-mediated autophagy (CMA), one of the main lysosomal degradative process, has a regulatory role in lipid metabolism of macrophages. However, whether the NLRP3 inflammasome is regulated by CMA, and the role of CMA in atherosclerosis remains unclear. OBJECTIVE: To determine the role of CMA in the regulation of NLRP3 inflammasome and atherosclerosis. METHODS AND RESULTS: The expression of CMA marker, LAMP-2A (lysosome-associated membrane protein type 2A), was first analyzed in ApoE-/- mouse aortas and human coronary atherosclerotic plaques, and a significant downregulation of LAMP-2A in advanced atherosclerosis in both mice and humans was observed. To selectively block CMA, we generated macrophage-specific conditional LAMP-2A knockout mouse strains in C57BL/6 mice and ApoE-/- mice. Deletion of macrophage LAMP-2A accelerated atherosclerotic lesion formation in the aortic root and the whole aorta in ApoE-/- mice. Mechanistically, LAMP-2A deficiency promoted NLRP3 inflammasome activation and subsequent release of mature IL (interleukin)-1ß in macrophages and atherosclerotic plaques. Furthermore, gain-of-function studies verified that restoration of LAMP-2A levels in LAMP-2A-deficient macrophages greatly attenuated NLRP3 inflammasome activation. Importantly, we identified the NLRP3 protein as a CMA substrate and demonstrated that LAMP-2A deficiency did not affect the NLRP3 mRNA levels but hindered degradation of the NLRP3 protein through CMA pathway. CONCLUSIONS: CMA function becomes impaired during the progression of atherosclerosis, which increases NLRP3 inflammasome activation and secretion of IL-1ß, promoting vascular inflammation and atherosclerosis progression. Our study unveils a new mechanism by which NLRP3 inflammasome is regulated in macrophages and atherosclerosis, thus providing a new insight into the role of autophagy-lysosomal pathway in atherosclerosis. Pharmacological activation of CMA may provide a novel therapeutic strategy for atherosclerosis and other NLRP3 inflammasome/IL-1ß-driven diseases.
Asunto(s)
Aterosclerosis/metabolismo , Autofagia , Inflamasomas/metabolismo , Proteína 2 de la Membrana Asociada a los Lisosomas/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Animales , Aorta/metabolismo , Aorta/patología , Apolipoproteínas E/genética , Apolipoproteínas E/metabolismo , Interleucina-1beta/metabolismo , Proteína 2 de la Membrana Asociada a los Lisosomas/genética , Macrófagos/metabolismo , Ratones , Ratones Endogámicos C57BL , Proteína con Dominio Pirina 3 de la Familia NLR/genéticaRESUMEN
Mirabegron (Myrbetriq) is a ß3-adrenoreceptor agonist approved for treating overactive bladder syndrome in human patients. This drug can activate brown adipose tissue (BAT) in adult humans and rodents through the ß3-adrenoreceptor-mediated sympathetic activation. However, the effect of the mirabegron, approved by the US Food and Drug Administration, on atherosclerosis-related cardiovascular disease is unknown. Here, we show that the clinical dose of mirabegron-induced BAT activation and browning of white adipose tissue (WAT) exacerbate atherosclerotic plaque development. In apolipoprotein E-/- (ApoE-/-) and low-density lipoprotein (LDL) receptor-/- (Ldlr-/-) mice, oral administration of clinically relevant doses of mirabegron markedly accelerates atherosclerotic plaque growth and instability by a mechanism of increasing plasma levels of both LDL-cholesterol and very LDL-cholesterol remnants. Stimulation of atherosclerotic plaque development by mirabegron is dependent on thermogenesis-triggered lipolysis. Genetic deletion of the critical thermogenesis-dependent protein, uncoupling protein 1, completely abrogates the mirabegron-induced atherosclerosis. Together, our findings suggest that mirabegron may trigger cardiovascular and cerebrovascular diseases in patients who suffer from atherosclerosis.
Asunto(s)
Acetanilidas/farmacología , Tejido Adiposo Pardo/efectos de los fármacos , Aterosclerosis/patología , Lipólisis/efectos de los fármacos , Tiazoles/farmacología , Agentes Urológicos/farmacología , Tejido Adiposo Pardo/metabolismo , Tejido Adiposo Blanco/efectos de los fármacos , Tejido Adiposo Blanco/metabolismo , Animales , Aterosclerosis/fisiopatología , LDL-Colesterol/sangre , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados para ApoE , Receptores de LDL/genéticaRESUMEN
The inflammation of adipose tissue is one of the most common secondary pathological changes in atherosclerosis, which in turn influences the process of atherosclerosis. Natriuretic peptides have been revealed important effect in regulating adipose metabolism. However, the relationship between natriuretic peptide receptor C and inflammation of adipose tissue in atherosclerosis remains unknown. This study aims to explore the effect natriuretic peptide receptor C exerts on the regulation of the adipose inflammation in atherosclerotic mice induced by western-type diet and its overlying mechanisms. To clarify the importance of NPRC of adipose inflammation in atherosclerotic mice, NPRC expression was measured in mice fed with chow diet and western-type diet for 12 weeks and we found a considerable increase in adipose tissue of atherosclerotic mice. Global NPRC knockout in mice was bred onto ApoE-/- mice to generate NPRC-/- ApoE-/- mice, which displayed remarked increase in browning of white adipose tissue and lipolysis of adipose tissue and decrease in adipose inflammation manifested by decreased macrophage invasion to form less CLS (crown-like structure), reduced oxidative stress and alleviated expression of TNFα, IL-6, IL-1ß and MCP1, but increased expression of adiponectin in adipose tissue. Moreover, our study showed that white adipose tissue browning in NPRC-/- ApoE-/- atherosclerotic mice was associated with decreased inflammatory response through cAMP/PKA signalling activation. These results identify NPRC as a novel regulator for adipose inflammation in atherosclerotic mice by modulating white adipose tissue browning.
Asunto(s)
Apolipoproteínas E/deficiencia , Hipercolesterolemia/complicaciones , Paniculitis/etiología , Paniculitis/metabolismo , Receptores del Factor Natriurético Atrial/deficiencia , Animales , Biomarcadores , AMP Cíclico , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Modelos Animales de Enfermedad , Susceptibilidad a Enfermedades , Regulación de la Expresión Génica , Hipercolesterolemia/genética , Hipercolesterolemia/metabolismo , Inmunohistoquímica , Inflamasomas/metabolismo , Ratones , Ratones Noqueados , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Estrés Oxidativo , Paniculitis/patología , Transducción de SeñalRESUMEN
Histone acetylation, an epigenetic modification, plays an important role in long-term memory formation. Recently, histone deacetylase (HDAC) inhibitors were demonstrated to promote memory formation, which raises the intriguing possibility that they may be used to rescue memory deficits. However, additional research is necessary to clarify the roles of individual HDACs in memory. In this study, we demonstrated that HDAC7, within the dorsal hippocampus of C57BL6J mice, had a late and persistent decrease after contextual fear conditioning (CFC) training (4-24 h), which was involved in long-term CFC memory formation. We also showed that HDAC7 decreased via ubiquitin-dependent degradation. CBX4 was one of the HDAC7 E3 ligases involved in this process. Nur77, as one of the target genes of HDAC7, increased 6-24 h after CFC training and, accordingly, modulated the formation of CFC memory. Finally, HDAC7 was involved in the formation of other hippocampal-dependent memories, including the Morris water maze and object location test. The current findings facilitate an understanding of the molecular and cellular mechanisms of HDAC7 in the regulation of hippocampal-dependent memory.SIGNIFICANCE STATEMENT The current findings demonstrated the effects of histone deacetylase 7 (HDAC7) on hippocampal-dependent memories. Moreover, we determined the mechanism of decreased HDAC7 in contextual fear conditioning (CFC) through ubiquitin-dependent protein degradation. We also verified that CBX4 was one of the HDAC7 E3 ligases. Finally, we demonstrated that Nur77, as one of the important targets for HDAC7, was involved in CFC memory formation. All of these proteins, including HDAC7, CBX4, and Nur77, could be potential therapeutic targets for preventing memory deficits in aging and neurological diseases.
Asunto(s)
Miedo/fisiología , Histona Desacetilasas/metabolismo , Ligasas/metabolismo , Memoria/fisiología , Complejo Represivo Polycomb 1/metabolismo , Ubiquitina-Proteína Ligasas/metabolismo , Ubiquitinación/fisiología , Animales , Condicionamiento Psicológico/fisiología , Miedo/psicología , Células HEK293 , Hipocampo/metabolismo , Humanos , Masculino , Aprendizaje por Laberinto/fisiología , Ratones , Ratones Endogámicos C57BLRESUMEN
Brain-derived neurotrophic factor (BDNF) plays an important role in neuronal survival, neurite outgrowth and synaptic plasticity by activating the receptor tropomyosin receptor kinase B (TrkB, also known as NTRK2). TrkB has been shown to undergo recycling after BDNF stimulation. We have previously reported that full-length TrkB (TrkB-FL) are recycled through a Rab11-dependent pathway upon BDNF stimuli, which is important for the translocation of TrkB-FL into dendritic spines and for the maintenance of prolonged BDNF downstream signaling during long-term potentiation (LTP). However, the identity of the motor protein that mediates the local transfer of recycled TrkB-FL back to the plasma membrane remains unclear. Here, we report that the F-actin-based motor protein myosin Va (Myo5a) mediates the postendocytic recycling of TrkB-FL. Blocking the interaction between Rab11 and Myo5a by use of a TAT-tagged peptide consisting of amino acids 55-66 of the Myo5a ExonE domain weakened the association between TrkB-FL and Myo5a and thus impaired TrkB-FL recycling and BDNF-induced TrkB-FL translocation into dendritic spines. Finally, inhibiting Myo5a-mediated TrkB-FL recycling led to a significant reduction in prolonged BDNF downstream signaling. Taken together, these results show that Myo5a mediates BDNF-dependent TrkB-FL recycling and contributes to BDNF-induced TrkB spine translocation and prolonged downstream signaling.
Asunto(s)
Factor Neurotrófico Derivado del Encéfalo/metabolismo , Espinas Dendríticas/metabolismo , Endocitosis/fisiología , Hipocampo/metabolismo , Cadenas Pesadas de Miosina/metabolismo , Miosina Tipo V/metabolismo , Neuronas/metabolismo , Receptor trkB/metabolismo , Animales , Western Blotting , Factor Neurotrófico Derivado del Encéfalo/genética , Células Cultivadas , Hipocampo/citología , Potenciación a Largo Plazo , Espectrometría de Masas , Ratones , Cadenas Pesadas de Miosina/genética , Miosina Tipo V/genética , Plasticidad Neuronal , Neuronas/citología , Transporte de Proteínas , Ratas , Receptor trkB/genética , Transducción de SeñalRESUMEN
Brain-derived neurotrophic factor (BDNF) plays an important role in the activity-dependent regulation of synaptic structure and function via tropomyosin related kinase B (TrkB) receptor activation. However, whether BDNF could regulate TrkB levels at synapse during long-term potentiation (LTP) is still unknown. We show in cultured rat hippocampal neurons that chemical LTP (cLTP) stimuli selectively promote endocytic recycling of BDNF-dependent full-length TrkB (TrkB-FL) receptors, but not isoform T1 (TrkB.T1) receptors, via a Rab11-dependent pathway. Moreover, neuronal-activity-enhanced TrkB-FL recycling could facilitate receptor translocation to postsynaptic density and enhance BDNF-induced extracellular signal-regulated kinase and phosphatidylinositol 3-kinase activation and rat hippocampal neuron survival. Finally, we found that cLTP could stimulate the switch of Rab11 from an inactive to an active form and that GTP-bound Rab11 could enhance the interaction between TrkB-FL and PSD-95. Therefore, the recycling endosome could serve as a reserve pool to supply TrkB-FL receptors for LTP maintenance. These findings provide a mechanistic link between Rab11-dependent endocytic recycling and TrkB modulation of synaptic plasticity.
Asunto(s)
Factor Neurotrófico Derivado del Encéfalo/metabolismo , Potenciación a Largo Plazo/fisiología , Neuronas/fisiología , Densidad Postsináptica/fisiología , Receptor trkB/metabolismo , Proteínas de Unión al GTP rab/metabolismo , Análisis de Varianza , Animales , Animales Recién Nacidos , Biotinilación , Homólogo 4 de la Proteína Discs Large , Estimulación Eléctrica , Embrión de Mamíferos , Femenino , Hipocampo/citología , Inmunoprecipitación , Etiquetado Corte-Fin in Situ , Técnicas In Vitro , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Potenciación a Largo Plazo/efectos de los fármacos , Masculino , Proteínas de la Membrana/metabolismo , Neuronas/ultraestructura , Técnicas de Placa-Clamp , Densidad Postsináptica/genética , Unión Proteica , Transporte de Proteínas/fisiología , Ratas , Ratas Sprague-Dawley , Fracciones Subcelulares/metabolismo , Proteínas de Unión al GTP rab/genéticaRESUMEN
At present, the association between circulating selenium and stroke is still in dispute. Thus, this study aimed to ascertain the association with a larger sample size than the previous study, based on the National Health and Nutrition Examination Survey (NHANES) from 2011 to 2018. In total, 13755 adults over the age of 20 years were included in our study. Multivariate logistic regression models were applied to analyze the correlation between blood selenium levels and stroke. The smooth curve fitting was performed to test the dose-response effects between blood selenium levels and stroke. After adjusting for all confounders, blood selenium levels were negatively associated with stroke (odds ratio [OR] = 0.57, 95% confidence interval [CI]: 0.37-0.87, P = 0.014). In the fully adjusted model, the highest tertile of blood selenium levels was negatively associated with stroke compared with the lowest tertile (OR = 0.70, 95% CI: 0.53-0.93, P for trend = 0.016). Moreover, the relationship between blood selenium levels and stroke was linear. In subgroup analyses, we observed that the test for interactions was significant for body mass index (BMI) and uric acid (P for interaction < 0.05). The negative relationship was stronger in participants with BMI 25-30 kg/m2 (OR = 0.23, 95% CI: 0.13-0.44, P < 0.001). Therefore, in American adults, the relationship between blood selenium levels and stroke was negative, with a linear tendency. In the future, a cohort study is warranted to further confirm this relationship.
Asunto(s)
Selenio , Accidente Cerebrovascular , Adulto , Humanos , Estados Unidos , Adulto Joven , Encuestas Nutricionales , Estudios de Cohortes , Índice de Masa CorporalRESUMEN
Our study aimed to examine whether whole blood selenium (WBSe) levels are related to the triglyceride-to-high-density lipoprotein cholesterol (TG/HDL-C) ratio among the general population. A total of 13,470 adults were included and analyzed from the National Health and Nutrition Examination Survey (NHANES) 2011-2018. In multivariable analyses, LnWBSe levels were significantly related to Ln(TG/HDL-C) ratio in fully adjusted model (ß = 0.35; 95% confidence interval (CI): 0.22, 0.48; P < 0.001). Furthermore, the highest quartile of LnWBSe levels was positively correlated with Ln(TG/HDL-C) ratio compared with the lowest quartile (ß = 0.15; 95% CI: 0.10, 0.20; P for trend < 0.001). In the dose-response analyses, the correlation was non-linear. While LnWBSe levels < 1.10, LnWBSe levels were positively related to Ln(TG/HDL-C) ratio (ß = 0.41; 95% CI: 0.31, 0.50; P < 0.001), whereas LnWBSe levels ≥ 1.10, the relationship was not significantly (ß = - 0.20; 95% CI: - 0.54, 0.13; P = 0.228). The interaction test was significant for age, sex, total cholesterol (TC), and diastolic blood pressure (DBP) (all P for interaction < 0.05). Overall, WBSe levels were positively related to TG/HDL-C ratio, with a non-linear trend. Further research is required to determine these underlying mechanisms.
Asunto(s)
Selenio , Adulto , Humanos , Triglicéridos , HDL-Colesterol , Encuestas Nutricionales , Presión SanguíneaRESUMEN
The lipid accumulation product (LAP) is a reliable marker of metabolic syndrome, which includes conditions like obesity. However, the correlation between the circulating selenium (CSe) concentration and the LAP is currently unclear. This study aimed to ascertain this correlation. Overall, 12,815 adults aged ≥20 years were enrolled in this study. After adjusting for all the confounding variables, CSe was positively correlated to the LAP (ß = 0.41; 95% confidence interval [CI]: 0.28, 0.54; p < 0.001). Compared with the lowest quartile of CSe, the highest quartile of CSe was positively related to the LAP (ß = 0.16; 95% CI: 0.12, 0.21; p < 0.001). Moreover, the correlation between CSe and the LAP revealed a positive non-linear trend. In the subgroup analysis, interaction effects were observed for age, sex, smoking, and stroke (p for interaction < 0.05). The effects were stronger for males (ß = 0.64, 95% CI: 0.47, 0.80; p < 0.001) and individuals who smoke at the time of the trial (ß = 0.64, 95% CI: 0.37, 0.91; p < 0.001). In conclusion, our results indicated that CSe was positively correlated with the LAP in a non-linear manner. Future research is warranted to explore their relationship and better understand the mechanisms underlying this association.
Asunto(s)
Producto de la Acumulación de Lípidos , Síndrome Metabólico , Selenio , Adulto , Masculino , Humanos , Estudios Transversales , Síndrome Metabólico/epidemiología , ObesidadRESUMEN
Background: The renin-angiotensin-aldosterone system (RAAS) members, especially Ang II and aldosterone, play key roles in the pathogenesis of diabetic cardiomyopathy (DCM). Angiotensin-converting enzyme inhibitors or angiotensin-receptor blockers combined with aldosterone receptor antagonists (mineralocorticoid receptor antagonists) have substantially improved clinical outcomes in patients with DCM. However, the use of the combination has been limited due to its high risk of inducing hyperkalemia. Methods: Type 1 diabetes was induced in 8-week-old male C57BL/6J mice by intraperitoneal injection of streptozotocin at a dose of 55 mg/kg for 5 consecutive days. Adeno-associated virus 9-mediated short-hairpin RNA (shRNA) was used to knock down the expression of ADAM17 in mice hearts. Eplerenone was administered via gavage at 200 mg/kg daily for 4 weeks. Primary cardiac fibroblasts were exposed to high glucose (HG) in vitro for 24 h to examine the cardiac fibroblasts to myofibroblasts transformation (CMT). Results: Cardiac collagen deposition and CMT increased in diabetic mice, leading to cardiac fibrosis and dysfunction. In addition, ADAM17 expression and activity increased in the hearts of diabetic mice. ADAM17 inhibition and eplerenone treatment both improved diabetes-induced cardiac fibrosis, cardiac hypertrophy and cardiac dysfunction, ADAM17 deficiency combined with eplerenone further reduced the effects of cardiac fibrosis, cardiac hypertrophy and cardiac dysfunction compared with single therapy in vivo. High-glucose stimulation promotes CMT in vitro and leads to increased ADAM17 expression and activity. ADAM17 knockdown and eplerenone pretreatment can reduce the CMT of fibroblasts that is induced by high glucose levels by inhibiting TGFß1/Smad3 activation; the combination of the two can further reduce CMT compared with single therapy in vitro. Conclusion: Our findings indicated that ADAM17 knockout could improve diabetes-induced cardiac dysfunction and remodeling through the inhibition of RAAS overactivation when combined with eplerenone treatment, which reduced TGF-ß1/Smad3 pathway activation-mediated CMT. The combined intervention of ADAM17 deficiency and eplerenone therapy provided additional cardiac protection compared with a single therapy alone without disturbing potassium level. Therefore, the combination of ADAM17 inhibition and eplerenone is a potential therapeutic strategy for human DCM.
RESUMEN
Abdominal aortic aneurysm (AAA) is a life-threatening vascular disease but effective drugs for treatment of AAA are still lacking. Recently, erythropoietin (EPO) is reported to induce AAA formation in apolipoprotein-E knock out (ApoE-/-) mice but an effective antagonist is unknown. In this study, formoterol, a ß2 adrenergic receptor (ß2AR) agonist, is found to be a promising agent for inhibiting AAA. To test this hypothesis, ApoE-/- mice are treated with vehicle, EPO, and EPO plus low-, medium-, and high-dose formoterol, respectively. The incidence of AAA is 0, 55%, 35%,10%, and 55% in these 5 groups, respectively. Mechanistically, senescence of vascular smooth muscle cell (VSMC) is increased by EPO while decreased by medium-dose formoterol both in vivo and in vitro, manifested by the altered expression of senescence biomarkers including phosphorylation of H2AXserine139, senescence-associated ß-galactosidase activity, and P21 protein level. In addition, expression of sirtuin 1 (SIRT1) in aorta is decreased in EPO-induced AAA but remarkably elevated by medium-dose formoterol. Knockdown of ß2AR and blockage of cyclic adenosine monophosphate (cAMP) attenuate the inhibitory role of formoterol in EPO-induced VSMC senescence. In summary, medium-dose formoterol attenuates EPO-induced AAA via ß2AR/cAMP/SIRT1 pathways, which provides a promising medication for the treatment of AAA.
Asunto(s)
Aneurisma de la Aorta Abdominal , Eritropoyetina , Fumarato de Formoterol , Animales , Ratones , Aneurisma de la Aorta Abdominal/inducido químicamente , Aneurisma de la Aorta Abdominal/tratamiento farmacológico , Apolipoproteínas E/metabolismo , Eritropoyetina/efectos adversos , Sirtuina 1/metabolismoRESUMEN
Laser speckle contrast imaging (LSCI) is widely used for in vivo real-time detection and analysis of local blood flow microcirculation due to its non-invasive ability and excellent spatial and temporal resolution. However, vascular segmentation of LSCI images still faces a lot of difficulties due to numerous specific noises caused by the complexity of blood microcirculation's structure and irregular vascular aberrations in diseased regions. In addition, the difficulties of LSCI image data annotation have hindered the application of deep learning methods based on supervised learning in the field of LSCI image vascular segmentation. To tackle these difficulties, we propose a robust weakly supervised learning method, which selects the threshold combinations and processing flows instead of labor-intensive annotation work to construct the ground truth of the dataset, and design a deep neural network, FURNet, based on UNet++ and ResNeXt. The model obtained from training achieves high-quality vascular segmentation and captures multi-scene vascular features on both constructed and unknown datasets with good generalization. Furthermore, we intravital verified the availability of this method on a tumor before and after embolization treatment. This work provides a new approach for realizing LSCI vascular segmentation and also makes a new application-level advance in the field of artificial intelligence-assisted disease diagnosis.
Asunto(s)
Inteligencia Artificial , Redes Neurales de la Computación , Rayos Láser , Microcirculación/fisiología , Aprendizaje Automático Supervisado , Procesamiento de Imagen Asistido por Computador/métodosRESUMEN
Selenium plays a role in obesity. However, whether circulating selenium levels are related to the visceral adiposity index (VAI), an indicator of obesity, is still unknown. Based on the National Health and Nutrition Examination Survey (NHANES) 2011-2018, data from 12,777 individuals aged ≥ 20 years were analyzed to clarify this question. In fully adjusted models of multivariate regression analysis, natural logarithm (Ln) selenium was positively related to Ln VAI (ß = 0.41; 95% confidence interval [CI], 0.27, 0.55; P < 0.001). Compared with the lowest quartile of Ln selenium, the highest quartile was also positively associated with Ln VAI (ß = 0.16; 95% CI, 0.11, 0.21; P < 0.001). Moreover, we found that this positive connection was non-linear. When Ln selenium was less than the inflection point, Ln selenium was positively related to Ln VAI (ß = 0.41; 95% CI, 0.31, 0.52; P < 0.001). However, when Ln selenium was greater than or equal to the inflection point, Ln selenium was not significantly related to Ln VAI (ß = -0.15; 95% CI, -0.56, 0.25; P = 0.455). In subgroup analysis, significant interactions were observed with age and sex (P for interaction < 0.05). Stronger interactions were observed among middle-aged participants (ß = 0.65; 95% CI, 0.31, 0.98; P = 0.002) and males (ß = 0.61; 95% CI, 0.43, 0.79; P < 0.001). Overall, circulating selenium levels were positively related to VAI, with a non-linear trend. Prospective studies and interventional experiments are necessary to verify the possible mechanisms.
RESUMEN
Abdominal aortic aneurysm (AAA) is a potentially fatal vascular disease, but the underlying mechanisms remain obscure. Here, we provide a protocol using erythropoietin (EPO) to induce the formation of AAA in both wild-type (WT) and apolipoprotein E (Apoe-/-) mice. We describe the dose, manner, and timing of EPO administration. We also detail mice dissection, aorta isolation, and histological analysis. The animal model of EPO-induced AAA provides a useful tool for exploring the mechanism of AAA in experimental studies. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2021).1.
Asunto(s)
Aneurisma de la Aorta Abdominal , Eritropoyetina , Animales , Ratones , Aorta Abdominal/patología , Modelos Animales de Enfermedad , Aneurisma de la Aorta Abdominal/inducido químicamente , Apolipoproteínas E/genética , Apolipoproteínas E/efectos adversos , Eritropoyetina/efectos adversosRESUMEN
Cardiac fibrosis plays a key role in the progression of diabetic cardiomyopathy (DCM). Previous studies demonstrated the cardioprotective effects of natriuretic peptides. However, the effects of natriuretic peptide receptor C (NPRC) on cardiac fibrosis in DCM remains unknown. Here, we observed that myocardial NPRC expression was increased in mice and patients with DCM. NPRC-/- diabetic mice showed alleviated cardiac fibrosis, as well as improved cardiac function and remodeling. NPRC knockdown in both cardiac fibroblasts and cardiomyocytes decreased collagen synthesis and proliferation of cardiac fibroblasts. RNA sequencing identified that NPRC deletion up-regulated the expression of TGF-ß-induced factor homeobox 1 (TGIF1), which inhibited the phosphorylation of Smad2/3. Furthermore, TGIF1 up-regulation was mediated by the activation of cAMP/PKA and cGMP/PKG signaling induced by NPRC deletion. These findings suggest that NPRC deletion attenuated cardiac fibrosis and improved cardiac remodeling and function in diabetic mice, providing a promising approach to the treatment of diabetic cardiac fibrosis.
Asunto(s)
Diabetes Mellitus Experimental , Cardiomiopatías Diabéticas , Receptores del Factor Natriurético Atrial , Animales , Ratones , Diabetes Mellitus Experimental/genética , Cardiomiopatías Diabéticas/genética , Cardiomiopatías Diabéticas/metabolismo , Fibrosis , Miocitos Cardíacos/metabolismo , Factor de Crecimiento Transformador beta1/genética , Factor de Crecimiento Transformador beta1/metabolismo , Receptores del Factor Natriurético Atrial/genéticaRESUMEN
Previous studies suggested a beneficial effect of natriuretic peptides in animal models of cardiovascular disease, but the role of natriuretic peptide receptor C (NPRC) in the pathogenesis of atherosclerosis (AS) remains unknown. This study was designed to test the hypothesis that NPRC may promote AS lesion formation and instability by enhancing oxidative stress, inflammation, and apoptosis via protein kinase A (PKA) signaling. ApoE-/- mice were fed chow or Western diet for 12 weeks and NPRC expression was significantly increased in the aortic tissues of Western diet-fed mice. Systemic NPRC knockout mice were crossed with ApoE-/- mice to generate ApoE-/-NPRC-/- mice, and NPRC deletion resulted in a significant decrease in the size and instability of aortic atherosclerotic lesions in ApoE-/-NPRC-/- versus ApoE-/- mice. In addition, endothelial cell-specific NPRC knockout attenuated atherosclerotic lesions in mice. In contrast, endothelial cell overexpression of NPRC aggravated the size and instability of atherosclerotic aortic lesions in mice. Experiments in vitro showed that NPRC knockdown in human aortic endothelial cells (HAECs) inhibited ROS production, pro-inflammatory cytokine expression and endothelial cell apoptosis, and increased eNOS expression. Furthermore, NPRC knockdown in HAECs suppressed macrophage migration, cytokine expression, and phagocytosis via its effects on endothelial cells. On the contrary, NPRC overexpression in endothelial cells resulted in opposite effects. Mechanistically, the anti-inflammation and anti-atherosclerosis effects of NPRC deletion involved activation of cAMP/PKA pathway, leading to downstream upregulated AKT1 pathway and downregulated NF-κB pathway. In conclusion, NPRC deletion reduced the size and instability of atherosclerotic lesions in ApoE-/- mice via attenuating inflammation and endothelial cell apoptosis and increasing eNOS expression by modulating cAMP/PKA-AKT1 and NF-κB pathways. Thus, targeting NPRC may provide a promising approach to the prevention and treatment of atherosclerosis.
Asunto(s)
Aterosclerosis , FN-kappa B , Receptores del Factor Natriurético Atrial , Animales , Humanos , Ratones , Apoptosis/genética , Aterosclerosis/genética , Aterosclerosis/metabolismo , Aterosclerosis/patología , Citocinas/metabolismo , Células Endoteliales/metabolismo , Inflamación/patología , Ratones Noqueados , Ratones Noqueados para ApoE , FN-kappa B/genética , FN-kappa B/metabolismo , Estrés Oxidativo/genética , Receptores del Factor Natriurético Atrial/genética , Receptores del Factor Natriurético Atrial/metabolismoRESUMEN
Myocardial infarction (MI) remains the leading cause of death worldwide. Cardiac fibroblasts (CFs) are abundant in the heart and are responsible for cardiac repair post-MI. NF-κB-repressing factor (NKRF) plays a significant role in the transcriptional inhibition of various specific genes. However, the NKRF action mechanism in CFs remains unclear in cardiac repair post-MI. This study investigates the NKRF mechanism in cardiac remodeling and dysfunction post-MI by establishing a CF-specific NKRF-knockout (NKRF-CKO) mouse model. NKRF expression is downregulated in CFs in response to pathological cardiac remodeling in vivo and TNF-α in vitro. NKRF-CKO mice demonstrate worse cardiac function and survival and increased infarct size, heart weight, and MMP2 and MMP9 expression post-MI compared with littermates. NKRF inhibits CF migration and invasion in vitro by downregulating MMP2 and MMP9 expression. Mechanistically, NKRF inhibits human antigen R (HuR) transcription by binding to the classical negative regulatory element within the HuR promoter via an NF-κB-dependent mechanism. This decreases HuR-targeted Mmp2 and Mmp9 mRNA stability. This study suggests that NKRF is a therapeutic target for pathological cardiac remodeling.
Asunto(s)
Infarto del Miocardio , FN-kappa B , Animales , Humanos , Ratones , Fibroblastos/metabolismo , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Infarto del Miocardio/metabolismo , FN-kappa B/metabolismo , Transducción de Señal/genética , Remodelación Ventricular/genéticaRESUMEN
Innate immunity is the first line to defend against pathogenic microorganisms, and Toll-like receptor (TLR)-mediated inflammatory responses are an essential component of innate immunity. However, the regulatory mechanisms of TLRs in innate immunity remain unperfected. We found that the expression of E3 ligase Ring finger protein 99 (RNF99) decreased significantly in peripheral blood monocytes from patients infected with Gram negative bacteria (G-) and macrophages stimulated by TLRs ligands, indicating the role of RNF99. We also demonstrated for the first time, the protective role of RNF99 against LPS-induced septic shock and dextran sodium sulfate (DSS)-induced colitis using RNF99 knockout mice (RNF99-/-) and bone marrow-transplanted mice. In vitro experiments revealed that RNF99 deficiency significantly promoted TLR-mediated inflammatory cytokine expression and activated the NF-κB and MAPK pathways in macrophages. Mechanistically, in both macrophages and HEK293 cell line with TLR4 stably transfection, RNF99 interacted with and degraded TAK1-binding protein (TAB) 2, a regulatory protein of the kinase TAK1, via the lysine (K)48-linked ubiquitin-proteasomal pathway on lysine 611 of TAB2, which further regulated the TLR-mediated inflammatory response. Overall, these findings indicated the physiological significance of RNF99 in macrophages in regulating TLR-mediated inflammatory reactions. It provided new insight into TLRs signal transduction, and offered a novel approach for preventing bacterial infections, endotoxin shock, and other inflammatory ills.