Repositorio Institucional Académico. Universidad Andrés Bello. Chile :: Examinando por Autor "Cabello-Verrugio, Claudio"

Examinando por Autor "Cabello-Verrugio, Claudio"

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Andrographolide attenuates skeletal muscle dystrophy in mdx mice and increases efficiency of cell therapy by reducing fibrosis
(BioMed Central Ltd., 2014-03) Cabrera, Daniel; Gutiérrez, Jaime; Cabello-Verrugio, Claudio; Morales, María G.; Mezzano, Sergio; Fadic, Ricardo; Casar, Juan C.; Hancke, Juan L.; Brandan, Enrique
Background: Duchenne muscular dystrophy (DMD) is characterized by the absence of the cytoskeletal protein dystrophin, muscle wasting, increased transforming growth factor type beta (TGF-β) signaling, and fibrosis. At the present time, the only clinically validated treatments for DMD are glucocorticoids. These drugs prolong muscle strength and ambulation of patients for a short term only and have severe adverse effects. Andrographolide, a bicyclic diterpenoid lactone, has traditionally been used for the treatment of colds, fever, laryngitis, and other infections with no or minimal side effects. We determined whether andrographolide treatment of mdx mice, an animal model for DMD, affects muscle damage, physiology, fibrosis, and efficiency of cell therapy. Methods: mdx mice were treated with andrographolide for three months and skeletal muscle histology, creatine kinase activity, and permeability of muscle fibers were evaluated. Fibrosis and TGF-β signaling were evaluated by indirect immunofluorescence and Western blot analyses. Muscle strength was determined in isolated skeletal muscles and by a running test. Efficiency of cell therapy was determined by grafting isolated skeletal muscle satellite cells onto the tibialis anterior of mdx mice. Results: mdx mice treated with andrographolide exhibited less severe muscular dystrophy than untreated dystrophic mice. They performed better in an exercise endurance test and had improved muscle strength in isolated muscles, reduced skeletal muscle impairment, diminished fibrosis and a significant reduction in TGF-β signaling. Moreover, andrographolide treatment of mdx mice improved grafting efficiency upon intramuscular injection of dystrophin-positive satellite cells. Conclusions: These results suggest that andrographolide could be used to improve quality of life in individuals with DMD.
Angiotensin-(1-7) improves skeletal muscle regeneration
(Page Press Publications, 2023) Valero-Breton, Mayalen; Tacchi, Franco; Abrigo, Johanna; Simon, Felipe; Cabrera, Daniel; Cabello-Verrugio, Claudio
Skeletal muscle possesses regenerative potential via satellite cells, compromised in muscular dystrophies leading to fibrosis and fat infiltration. Angiotensin II (Ang-II) is commonly associated with pathological states. In contrast, Angiotensin (1-7) [Ang-(1-7)] counters Ang-II, acting via the Mas receptor. While Ang-II affects skeletal muscle regeneration, the influence of Ang-(1-7) remains to be elucidated. Therefore, this study aims to investigate the role of Ang-(1-7) in skeletal muscle regeneration. C2C12 cells were differentiated in the absence or presence of 10 nM of Ang-(1-7). The diameter of myotubes and protein levels of myogenin and myosin heavy chain (MHC) were determined. C57BL/6 WT male mice 16-18 weeks old) were randomly assigned to injury-vehicle, injury-Ang-(1-7), and control groups. Ang-(1-7) was administered via osmotic pumps, and muscle injury was induced by injecting barium chloride to assess muscle regeneration through histological analyses. Moreover, embryonic myosin (eMHC) and myogenin protein levels were evaluated. C2C12 myotubes incubated with Ang-(1-7) showed larger diameters than the untreated group and increased myogenin and MHC protein levels during differentiation. Ang-(1-7) administration enhances regeneration by promoting a larger diameter of new muscle fibers. Furthermore, higher numbers of eMHC (+) fibers were observed in the injured-Ang-(1-7), which also had a larger diameter. Moreover, eMHC and myogenin protein levels were elevated, supporting enhanced regeneration due to Ang-(1-7) administration. Ang-(1-7) effectively promotes differentiation in vitro and improves muscle regeneration in the context of injuries, with potential implications for treating muscle-related disorders. © 2023 PAGEPress Publications. All rights reserved.
Cholic and deoxycholic acids induce mitochondrial dysfunction, impaired biogenesis and autophagic flux in skeletal muscle cells
(BioMed Central Ltd, 2023-06) Abrigo, Johanna; Olguín, Hugo; Tacchi, Franco; Orozco-Aguilar, Josué; Valero-Breton, Mayalen; Soto, Jorge; Castro-Sepúlveda, Mauricio; Elorza, Alvaro A.; Simon, Felipe; Cabello-Verrugio, Claudio
Background: Skeletal muscle is sensitive to bile acids (BA) because it expresses the TGR5 receptor for BA. Cholic (CA) and deoxycholic (DCA) acids induce a sarcopenia-like phenotype through TGR5-dependent mechanisms. Besides, a mouse model of cholestasis-induced sarcopenia was characterised by increased levels of serum BA and muscle weakness, alterations that are dependent on TGR5 expression. Mitochondrial alterations, such as decreased mitochondrial potential and oxygen consumption rate (OCR), increased mitochondrial reactive oxygen species (mtROS) and unbalanced biogenesis and mitophagy, have not been studied in BA-induced sarcopenia. Methods: We evaluated the effects of DCA and CA on mitochondrial alterations in C2C12 myotubes and a mouse model of cholestasis-induced sarcopenia. We measured mitochondrial mass by TOM20 levels and mitochondrial DNA; ultrastructural alterations by transmission electronic microscopy; mitochondrial biogenesis by PGC-1α plasmid reporter activity and protein levels by western blot analysis; mitophagy by the co-localisation of the MitoTracker and LysoTracker fluorescent probes; mitochondrial potential by detecting the TMRE probe signal; protein levels of OXPHOS complexes and LC3B by western blot analysis; OCR by Seahorse measures; and mtROS by MitoSOX probe signals. Results: DCA and CA caused a reduction in mitochondrial mass and decreased mitochondrial biogenesis. Interestingly, DCA and CA increased LC3II/LC3I ratio and decreased autophagic flux concordant with raised mitophagosome-like structures. In addition, DCA and CA decreased mitochondrial potential and reduced protein levels in OXPHOS complexes I and II. The results also demonstrated that DCA and CA decreased basal, ATP-linked, FCCP-induced maximal respiration and spare OCR. DCA and CA also reduced the number of cristae. In addition, DCA and CA increased the mtROS. In mice with cholestasis-induced sarcopenia, TOM20, OXPHOS complexes I, II and III, and OCR were diminished. Interestingly, the OCR and OXPHOS complexes were correlated with muscle strength and bile acid levels. Conclusion: Our results showed that DCA and CA decreased mitochondrial mass, possibly by reducing mitochondrial biogenesis, which affects mitochondrial function, thereby altering potential OCR and mtROS generation. Some mitochondrial alterations were also observed in a mouse model of cholestasis-induced sarcopenia characterised by increased levels of BA, such as DCA and CA. © 2023, The Author(s).
Correction for Echeverría et al., "Endotoxin-Induced Endothelial Fibrosis Is Dependent on Expression of Transforming Growth Factors β1 and β2" [Infect Immun., Volume 82, no. 9, p. 3678-3686, 2014]
(American Society for Microbiology, 2015) Echeverría, César; Montorfano, Ignacio; Tapia, Pablo; Riedel, Claudia; Cabello-Verrugio, Claudio; Simon, Felipe a
Desarrollo de la cultura del cuidado mediante la implementación de un programa en el uso de animales de laboratorio
(Universidad Andrés Bello, 2021) Valdivia Jeldres, Rebeca Alejandra; Valenzuela Anguita, Sebastián Andrés; Cabello-Verrugio, Claudio; Facultad de Ciencias de la Vida
La Cultura del Cuidado es un concepto novedoso, que describe el compromiso con el mejoramiento, la calidad, el cuidado y la transparencia hacia el bienestar animal, el cual es promovido a través de una mentalidad y un comportamiento que trasciende el cumplimiento de normas y regulaciones. Esta visión revoluciona el manejo, cuidado y uso de animales de laboratorio, porque busca fortalecer la interacciones humano-humano y animal-humano, promover el juicio crítico de las personas respecto a la reducción del daño y el refinamiento de los procedimientos experimentales, enmarcando los valores ideales para generar una mayor empatía y conexión emocional, mejorando el bienestar de los animales y la obtención de datos en las investigaciones científicas. Es por esto que el objetivo de esta investigación fue implementar un Programa Institucional de Cuidado y Uso de Animales y observar si se lograba un reconocimiento y una comprensión de la Cultura del Cuidado en una institución científica. Se aplicó una encuesta antes y después de la realización del programa de forma voluntaria a los participantes, con los datos se evaluó el impacto que tuvo el programa, mediante dos análisis de Anova, que midieron la eficacia del programa y las posibles incidencias de las variables categóricas en el modelo experimental. Los resultados obtenidos reflejaron una diferencia significativa en el efecto que tuvo el programa en los participantes (p = 4.545e06), y una incidencia significativa de la variable “Profesión” en el reconocimiento de la Cultura del Cuidado en el modelo experimental. Estos resultados validan el modelo propuesto y apoyan los trabajos realizados previamente, sugiriendo que la implementación de un modelo educativo que tenga como base la Cultura del Cuidado, fortalece el reconocimiento de este nuevo término y genera cambios significativos en la estructura ética y moral de lo trabajadores que utilizan animales con fines científicos.
Effect of Dietary Supplements with ω-3 Fatty Acids, Ascorbic Acid, and Polyphenolic Antioxidant Flavonoid on Gene Expression, Organ Failure, and Mortality in Endotoxemia-Induced Septic Rats
(MDPI, 2023-03) Prado, Yolanda; Echeverría, Cesar; Feijóo, Carmen G.; Riedel, Claudia A.; Cabello-Verrugio, Claudio; Santibanez, Juan F.; Simon, Felipe
Sepsis syndrome develops through enhanced secretion of pro-inflammatory cytokines and the generation of reactive oxygen species (ROS). Sepsis syndrome is characterized by vascular hyperpermeability, hypotension, multiple organ dysfunction syndrome (MODS), and increased mortality, among others. Endotoxemia-derived sepsis is an important cause of sepsis syndrome. During endotoxemia, circulating endotoxin interacts with endothelial cells (ECs), inducing detrimental effects on endothelium function. The endotoxin induces the conversion of ECs into fibroblasts, which are characterized by a massive change in the endothelial gene-expression pattern. This downregulates the endothelial markers and upregulates fibrotic proteins, mesenchymal transcription factors, and extracellular matrix proteins, producing endothelial fibrosis. Sepsis progression is modulated by the consumption of specific nutrients, including ω-3 fatty acids, ascorbic acid, and polyphenolic antioxidant flavonoids. However, the underlying mechanism is poorly described. The notion that gene expression is modulated during inflammatory conditions by nutrient consumption has been reported. However, it is not known whether nutrient consumption modulates the fibrotic endothelial gene-expression pattern during sepsis as a mechanism to decrease vascular hyperpermeability, hypotension, MODS, and mortality. Therefore, the aim of this study was to investigate the impact of the consumption of dietary ω-3 fatty acids, ascorbic acid, and polyphenolic antioxidant flavonoid supplements on the modulation of fibrotic endothelial gene-expression patterns during sepsis and to determine the effects on sepsis outcomes. Our results indicate that the consumption of supplements based on ω-3 fatty acids and polyphenolic antioxidant flavonoids was effective for improving endotoxemia outcomes through prophylactic ingestion and therapeutic usage. Thus, our findings indicated that specific nutrient consumption improves sepsis outcomes and should be considered in treatment. © 2023 by the authors.
Endothelial dysfunction in pregnancy metabolic disorders
(Elsevier B.V., 2020-02) Echeverria, Cesar; Eltit, Felipe; Santibanez, Juan F; Gatica, Sebastian; Cabello-Verrugio, Claudio; Simon, Felipe
In recent years, the vascular endothelium has gained attention as a key player in the initiation and development of pregnancy disorders. Endothelium acts as an endocrine organ that preserves the homeostatic balance by responding to changes in metabolic status. However, in metabolic disorders, endothelial cells adopt a dysfunctional function, losing their normal responsiveness. During pregnancy, several metabolic changes occur, in which endothelial function decisively participates. Similarly, when pregnancy metabolic disorders occur, endothelial dysfunction plays a key role in pathogenesis. This review outlines the main findings regarding endothelial dysfunction in three main metabolic pathological conditions observed during pregnancy: gestational diabetes, hypertensive disorders, and obesity and hyperlipidemia. Organ, histological and cellular characteristics were thoroughly described. Also, we focused in discussing the underlying molecular mechanisms involved in the cellular signaling pathways that mediate responses in these pathological conditions. © 2019 Elsevier B.V.
Endothelial fibrosis induced by suppressed STAT3 expression mediated by signaling involving the TGF-β1/ALK5/Smad pathway
(Nature Publishing Group, 2017-09) Becerra, Alvaro; Rojas, MacArena; Vallejos, Alejandro; Villegas, Vicente; Pérez, Lorena; Cabello-Verrugio, Claudio; Simon, Felipe
During systemic inflammatory pathologies, mediators of inflammation circulate in the bloodstream and interact with endothelial cells (ECs), resulting in endothelial dysfunction that maintains and enhances the pathological condition. Inflammatory mediators change the protein expression profile of ECs, which become activated fibroblasts via endothelial-to-mesenchymal transition. This process is characterized by downregulated endothelial proteins and strongly upregulated fibrotic-specific genes and extracellular matrix-forming proteins. The main inductor of endothelial fibrosis is transforming growth factor-β1 (TGF-β1), which acts through the TGF-β1/activin receptor-like kinase 5 (ALK5)/Smads intracellular signaling pathway. The signal transducer and activator of transcription 3 (STAT3) is also involved in fibrosis in several tissues (e.g. heart and vascular system), where STAT3 signaling decreases TGF-β1-induced responses by directly interacting with Smad proteins, suggesting that decreased STAT3 could induce TGF-β1-mediated fibrosis. However, it is unknown if suppressed STAT3 expression induces EC fibrosis through a mechanism involving the TGF-β signaling pathway. The present study evaluated the fibrotic actions of STAT3 suppression in ECs and investigated TGF-β1/ALK5/Smad4 signaling pathway participation. Suppressed STAT3 expression stimulated fibrotic conversion in ECs, as mediated by protein expression reprograming that decreased endothelial marker expression and increased fibrotic and extracellular matrix protein levels. The potential mechanism underlying these changes was dependent on TGF-β1 secretion, the ALK5 activation pathway, and Smad4 translocation into the nucleus. We conclude that suppressed STAT3 expression converts ECs into activated fibroblasts via TGF-β1/ALK5/Smad4 signaling pathway involvement. © 2017 USCAP, Inc.
Endotoxin induces fibrosis in vascular endothelial cells through a mechanism dependent on transient receptor protein melastatin 7 activity
(Public Library of Science, 2014-04) Echeverría, Cesar; Montorfano, Ignacio; Hermosilla, Tamara; Armisén, Ricardo; Velásquez, Luis A.; Cabello-Verrugio, Claudio; Varela, Diego; Simon, Felipe
The pathogenesis of systemic inflammatory diseases, including endotoxemia-derived sepsis syndrome, is characterized by endothelial dysfunction. It has been demonstrated that the endotoxin lipopolysaccharide (LPS) induces the conversion of endothelial cells (ECs) into activated fibroblasts through endothelial-to-mesenchymal transition mechanism. Fibrogenesis is highly dependent on intracellular Ca2+ concentration increases through the participation of calcium channels. However, the specific molecular identity of the calcium channel that mediates the Ca2+ influx during endotoxin-induced endothelial fibrosis is still unknown. Transient receptor potential melastatin 7 (TRPM7) is a calcium channel that is expressed in many cell types, including ECs. TRPM7 is involved in a number of crucial processes such as the conversion of fibroblasts into activated fibroblasts, or myofibroblasts, being responsible for the development of several characteristics of them. However, the role of the TRPM7 ion channel in endotoxin-induced endothelial fibrosis is unknown. Thus, our aim was to study whether the TRPM7 calcium channel participates in endotoxin-induced endothelial fibrosis. Using primary cultures of ECs, we demonstrated that TRPM7 is a crucial protein involved in endotoxin-induced endothelial fibrosis. Suppression of TRPM7 expression protected ECs from the fibrogenic process stimulated by endotoxin. Downregulation of TRPM7 prevented the endotoxin-induced endothelial markers decrease and fibrotic genes increase in ECs. In addition, TRPM7 downregulation abolished the endotoxin-induced increase in ECM proteins in ECs. Furthermore, we showed that intracellular Ca2+ levels were greatly increased upon LPS challenge in a mechanism dependent on TRPM7 expression. These results demonstrate that TRPM7 is a key protein involved in the mechanism underlying endotoxin-induced endothelial fibrosis.
Endotoxin-induced endothelial fibrosis is dependent on expression of transforming growth factors β1 and β2
(American Society for Microbiology, 2014) Echeverría, César; Montorfano, Ignacio; Tapia, Pablo; Riedel, Claudia; Cabello-Verrugio, Claudio; Simon, Felipe
During endotoxemia-induced inflammatory disease, bacterial endotoxins circulate in the bloodstream and interact with endo thelial cells (ECs), inducing dysfunction of the ECs. We previously reported that endotoxins induce the conversion of ECs into activated fibroblasts. Through endotoxin-induced endothelial fibrosis, ECs change their morphology and their protein expres sion pattern, thereby suppressing endothelial markers and upregulating fibrotic proteins. The most commonly used fibrotic in ducers are transforming growth factor 1 (TGF- 1) and TGF- 2. However, whether TGF- 1 and TGF- 2 participate in endo toxin-induced endothelial fibrosis remains unknown. We have shown that the endotoxin-induced endothelial fibrosis process is dependent on the TGF- receptor, ALK5, and the activation of Smad3, a protein that is activated by ALK5 activation, thus sug gesting that endotoxin elicits TGF- production to mediate endotoxin-induced endothelial fibrosis. Therefore, we investigated the dependence of endotoxin-induced endothelial fibrosis on the expression of TGF- 1 and TGF- 2. Endotoxin-treated ECs induced the expression and secretion of TGF- 1 and TGF- 2. TGF- 1 and TGF- 2 downregulation inhibited the endotoxin induced changes in the endothelial marker VE-cadherin and in the fibrotic proteins -SMA and fibronectin. Thus, endotoxin in duces the production of TGF- 1 and TGF- 2 as a mechanism to promote endotoxin-induced endothelial fibrosis. To the best of our knowledge, this is the first report showing that endotoxin induces endothelial fibrosis via TGF- secretion, which represents an emerging source of vascular dysfunction. These findings contribute to understanding the molecular mechanism of endotox in-induced endothelial fibrosis, which could be useful in the treatment of inflammatory diseases
Entrenamiento físico previene la sarcopenia inducida por enfermedad hepática crónica
(Universidad Andrés Bello, 2023) González Rojas, Andrea Patricia; Cabello-Verrugio, Claudio
La enfermedad hepática crónica (EHC) produce sarcopenia secundaria a la disfunción del hígado, que se caracteriza por disminución de la fuerza muscular, masa muscular y función física y es un predictor independiente de mortalidad pre y post trasplante hepático. El entrenamiento físico de resistencia (ER), fuerza (EF) o entrenamiento combinado (EC), puede ser una herramienta no farmacológica que prevenga y trate la sarcopenia en EHC, sin embargo, no existen estudios comparativos al respecto. La hipótesis planteada es que la sarcopenia secundaria a EHC es prevenida en mayor grado por el EC, respecto al EF o ER, en un modelo preclínico. El objetivo es comparar el efecto de los entrenamientos de fuerza, resistencia y EC, sobre la sarcopenia inducida por EHC en un modelo murino. Ratones C57BL6 sanos (controles) y con EHC, se subdividieron en grupos sedentarios (SED), EF, ER y EC. La intervención experimental con dieta y entrenamiento se realizó por 6 semanas, este último a intensidad moderada-vigorosa. Se evaluó fuerza muscular in vivo y ex vivo, masa muscular general, masa de músculos, diámetro de fibras y niveles de proteínas Troponina, cadena pesada de la miosina y p70S6K total, y función física de alto, moderado y bajo esfuerzo. Los resultados indican que, durante la EHC, la pérdida de fuerza in vivo y ex vivo, es prevenida por los tres tipos de entrenamiento físico. La pérdida de masa muscular general no es prevenida por el entrenamiento físico en animales con EHC, pero sí existe un efecto preventivo de los tres entrenamientos sobre la disminución de la masa de los músculos, el diámetro de las fibras musculares del TA y SOL y el nivel de las proteínas analizadas. La pérdida de la función física es prevenida en mayor medida por el ER y EC. En conjunto, los datos nos permiten concluir que el EF, ER y EC tienen efectos preventivos sobre la disminución de la fuerza, la masa muscular y la función física que caracterizan la EHC. El EC podría ser una buena opción para tratar y prevenir la sarcopenia en EHC, respecto al EF y el ER por sí solos.
Erratum for: Transforming growth factor type-β inhibits Mas receptor expression in fibroblasts but not in myoblasts or differentiated myotubes; Relevance to fibrosis associated to muscular dystrophies [BioFactors, 2015 March/April; 42(2); 111-120, doi: 10.1002/biof.1208]
(BIFAE, 2015-05) Cofre, Catalina; José Acuña, María; Contreras, Osvaldo; Gabriela Morales, María; Riquelme, Cecilia; Bader, Michael; Santos, Robson; Cabello-Verrugio, Claudio; Brandan, Enrique
High Fat Diet-Induced Skeletal Muscle Wasting Is Decreased by Mesenchymal Stem Cells Administration: Implications on Oxidative Stress, Ubiquitin Proteasome Pathway Activation, and Myonuclear Apoptosis
(HINDAWI PUBLISHING CORP, 2016-06) Abrigo, Johanna; Rivera, Juan Carlos; Aravena, Javier; Cabrera, Daniel; Simon, Felipe; Ezquer, Fernando; Ezquer, Marcelo; Cabello-Verrugio, Claudio
Obesity can lead to skeletal muscle atrophy, a pathological condition characterized by the loss of strength and muscle mass. A feature of muscle atrophy is a decrease of myofibrillar proteins as a result of ubiquitin proteasome pathway overactivation, as evidenced by increased expression of the muscle-specific ubiquitin ligases atrogin-1 and MuRF-1. Additionally, other mechanisms are related to muscle wasting, including oxidative stress, myonuclear apoptosis, and autophagy. Stem cells are an emerging therapy in the treatment of chronic diseases such as high fat diet-induced obesity. Mesenchymal stem cells (MSCs) are a population of self-renewable and undifferentiated cells present in the bone marrow and other mesenchymal tissues of adult individuals. The present study is the first to analyze the effects of systemic MSC administration on high fat diet-induced skeletal muscle atrophy in the tibialis anterior of mice. Treatment with MSCs reduced losses of muscle strength and mass, decreases of fiber diameter and myosin heavy chain protein levels, and fiber type transitions. Underlying these antiatrophic effects, MSC administration also decreased ubiquitin proteasome pathway activation, oxidative stress, and myonuclear apoptosis. These results are the first to indicate that systemically administered MSCs could prevent muscle wasting associated with high fat diet-induced obesity and diabetes.
Intensive care unit-acquired weakness: a review from molecular mechanisms to its impact in COVID-2019
(Page Press Publications, 2022-09) González, Andrea; Abrigo, Johanna; Achiardi, Oscar; Simon, Felipe; Cabello-Verrugio, Claudio
Intensive Care Unit-Acquired Weakness (ICU-AW) is a generalized and symmetric neuromuscular dysfunction associated with critical illness and its treatments. Its incidence is approximately 80% in intensive care unit patients, and it manifests as critical illness polyneuropathy, critical illness myopathy, and muscle atrophy. Intensive care unit patients can lose an elevated percentage of their muscle mass in the first days after admission, producing short- and long-term sequelae that affect patients’ quality of life, physical health, and mental health. In 2019, the world was faced with coronavirus disease 2019 (COVID-19), caused by the acute respiratory syndrome coronavirus 2. COVID-19 produces severe respiratory disorders, such as acute respiratory distress syndrome, which increases the risk of developing ICU-AW. COVID-19 patients treated in intensive care units have shown early diffuse and symmetrical muscle weakness, polyneuropathy, and myalgia, coinciding with the clinical presentation of ICU-AW. Besides, these patients require prolonged intensive care unit stays, invasive mechanical ventilation, and intensive care unit pharmacological therapy, which are risk factors for ICU-AW. Thus, the purposes of this review are to discuss the features of ICU-AW and its effects on skeletal muscle. Further, we will describe the mechanisms involved in the probable development of ICU-AW in severe COVID-19 patients.
NEDD4-1 deficiency impairs satellite cell function during skeletal muscle regeneration
(BioMed Central Ltd., 2023-05) Cabezas, Felipe; Cabello-Verrugio, Claudio; González, Natalia; Salas, Jeremy; Ramírez, Manuel J.; De la Vega, Eduardo; Olguín, Hugo C.
Background: Satellite cells are tissue-specific stem cells primarily responsible for the regenerative capacity of skeletal muscle. Satellite cell function and maintenance are regulated by extrinsic and intrinsic mechanisms, including the ubiquitin–proteasome system, which is key for maintaining protein homeostasis. In this context, it has been shown that ubiquitin-ligase NEDD4-1 targets the transcription factor PAX7 for proteasome-dependent degradation, promoting muscle differentiation in vitro. Nonetheless, whether NEDD4-1 is required for satellite cell function in regenerating muscle remains to be determined. Results: Using conditional gene ablation, we show that NEDD4-1 loss, specifically in the satellite cell population, impairs muscle regeneration resulting in a significant reduction of whole-muscle size. At the cellular level, NEDD4-1-null muscle progenitors exhibit a significant decrease in the ability to proliferate and differentiate, contributing to the formation of myofibers with reduced diameter. Conclusions: These results indicate that NEDD4-1 expression is critical for proper muscle regeneration in vivo and suggest that it may control satellite cell function at multiple levels. © 2023, The Author(s).
OxHDL controls LOX-1 expression and plasma membrane localization through a mechanism dependent on NOX/ROS/NF-κB pathway on endothelial cells
(Nature Publishing Group, 2019-03) Pérez, Lorena; Vallejos, Alejandro; Echeverria, Cesar; Varela, Diego; Cabello-Verrugio, Claudio; Simon, Felipe
Systemic inflammatory diseases enhance circulating oxidative stress levels, which results in the oxidation of circulating high-density lipoprotein (oxHDL). Endothelial cell function can be negatively impacted by oxHDL, but the underlying mechanisms for this remain unclear. Some reports indicate that the lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is also a receptor for oxHDL. However, it is unknown if oxHDL induces increased LOX-1 expression at the plasma membrane, as an event that supports endothelial dysfunction. Therefore, the aims of this study were to determine if oxHDL induces plasma-membrane level changes in LOX-1 and, if so, to describe the underlying mechanisms in endothelial cells. Our results demonstrate that the incubation of arterial or vein endothelial cells with oxHDL (and not HDL) induces the increase of LOX-1 expression at the plasma membrane; effect prevented by LOX-1 inhibition. Importantly, same results were observed in endothelial cells from oxHDL-treated rats. Furthermore, the observed oxHDL-induced LOX-1 expression is abolished by the down-regulation of NOX-2 expression with siRNA (and no others NOX isoforms), by the pharmacological inhibition of NAD(P)H oxidase (with DPI or apocynin) or by the inhibition of NF-κB transcription factor. Coherently, LOX-1 expression is augmented by the incubation of endothelial cells with H2O2 or GSSG even in absence of oxHDL, indicating that the NOX-2/ROS/ NF-κB axis is involved. Interestingly, oxHDL incubation also increases TNF-α expression, cytokine that induces LOX-1 expression. Thus, our results suggest a positive feedback mechanism for LOX-1 receptor during inflammatory condition where an oxidative burst will generate oxHDL from native HDL, activating LOX-1 receptor which in turn will increase the expression of NOX-2, TNF-α and LOX-1 receptor at the plasma membrane. In conclusion, oxHDL-induced translocation of LOX-1 to the plasma membrane could constitute an induction mechanism of endothelial dysfunction in systemic inflammatory diseases. © 2019, United States & Canadian Academy of Pathology.
OxHDL controls LOX-1 expression and plasma membrane localization through a mechanism dependent on NOX/ROS/NF-κB pathway on endothelial cells
(Nature Publishing Group, 2019-03-01) Pérez, Lorena; Vallejos, Alejandro; Echeverria, Cesar; Varela, Diego; Cabello-Verrugio, Claudio; Simon, Felipe
Systemic inflammatory diseases enhance circulating oxidative stress levels, which results in the oxidation of circulating high-density lipoprotein (oxHDL). Endothelial cell function can be negatively impacted by oxHDL, but the underlying mechanisms for this remain unclear. Some reports indicate that the lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is also a receptor for oxHDL. However, it is unknown if oxHDL induces increased LOX-1 expression at the plasma membrane, as an event that supports endothelial dysfunction. Therefore, the aims of this study were to determine if oxHDL induces plasma-membrane level changes in LOX-1 and, if so, to describe the underlying mechanisms in endothelial cells. Our results demonstrate that the incubation of arterial or vein endothelial cells with oxHDL (and not HDL) induces the increase of LOX-1 expression at the plasma membrane; effect prevented by LOX-1 inhibition. Importantly, same results were observed in endothelial cells from oxHDL-treated rats. Furthermore, the observed oxHDL-induced LOX-1 expression is abolished by the down-regulation of NOX-2 expression with siRNA (and no others NOX isoforms), by the pharmacological inhibition of NAD(P)H oxidase (with DPI or apocynin) or by the inhibition of NF-κB transcription factor. Coherently, LOX-1 expression is augmented by the incubation of endothelial cells with H2O2 or GSSG even in absence of oxHDL, indicating that the NOX-2/ROS/ NF-κB axis is involved. Interestingly, oxHDL incubation also increases TNF-α expression, cytokine that induces LOX-1 expression. Thus, our results suggest a positive feedback mechanism for LOX-1 receptor during inflammatory condition where an oxidative burst will generate oxHDL from native HDL, activating LOX-1 receptor which in turn will increase the expression of NOX-2, TNF-α and LOX-1 receptor at the plasma membrane. In conclusion, oxHDL-induced translocation of LOX-1 to the plasma membrane could constitute an induction mechanism of endothelial dysfunction in systemic inflammatory diseases.
Oxidized High-Density Lipoprotein Induces Endothelial Fibrosis Promoting Hyperpermeability, Hypotension, and Increased Mortality
(Antioxidants, 2022) Rojas, Macarena; Prado, Yolanda; Tapia, Pablo; Carreño, Leandro J.; Cabello-Verrugio, Claudio; Simon, Felipe
During systemic inflammation, reactive oxygen species (ROS) are generated in the blood-stream, producing large amounts of oxidized HDL (oxHDL). OxHDL loses the vascular protective features of native HDL, acquiring detrimental actions. Systemic inflammation promotes endothelial fibrosis, characterized by adhesion protein downregulation and fibrotic-specific gene upregulation, disrupting endothelial monolayer integrity. Severe systemic inflammatory conditions, as found in critically ill patients in the intensive care unit (ICU), exhibit endothelial hyperpermeability, hypoten sion, and organ hypoperfusion, promoting organ dysfunction and increased mortality. Because endothelial fibrosis disturbs the endothelium, it is proposed that it is the cellular and molecular origin of endothelial hyperpermeability and the subsequent deleterious consequences. However, whether oxHDL is involved in this process is unknown. The aim of this study was to investigate the fibrotic effect of oxHDL on the endothelium, to elucidate the underlying molecular and cellular mechanism, and to determine its effects on vascular permeability, blood pressure, and mortality. The results showed that oxHDL induces endothelial fibrosis through the LOX-1/NOX-2/ROS/NF-κB pathway, TGF-β secretion, and ALK-5/Smad activation. OxHDL-treated rats showed endothelial hyperpermeability, hypotension, and an enhanced risk of death and mortality, which was prevented using an ALK-5 inhibitor and antioxidant diet consumption. Additionally, the ICU patients showed fibrotic endothelial cells, and the resuscitation fluid volume administered correlated with the plasma oxHDL levels associated with an elevated risk of death and mortality. We conclude that oxHDL generates endothelial fibrosis, impacting blood pressure regulation and survival.
Regulación de la actividad biológica dependiente de miostatina por Angiotensina (1-7)
(Universidad Andrés Bello, 2016) Aravena Castro, Javier Ignacio; Cabello-Verrugio, Claudio; Facultad de Ciencias Biológicas; Escuela de Ingeniería en Biotecnología
La atrofia muscular esquelética está caracterizada por la pérdida de la masa muscular y la fuerza, principalmente debido a un incremento en la degradación de proteínas miofibrilares, como la cadena pesada de miosina (MHC). Miostatina (MSTN) es un factor proteico soluble que regula la masa muscular y cuyos niveles están incrementados en varios tipos de atrofia muscular. Dentro de los efectos de MSTN en atrofia, se encuentra un aumento de la degradación de proteínas miofibrilares debido a una mayor actividad del sistema ubiquitina-proteosoma (UPS), especialmente un incremento de Atrogina-1 y MuRF-1, dos E3 ubiquitina-ligasas. El efecto atrófico inducido por MSTN es dependiente de la activación de la vía de señalización de Smad. El sistema renina angiotensina (RAS) es un regulador de la masa muscular. Angiotensina (1-7) (Ang (1-7)) es un péptido del eje no-clásico de RAS. En nuestro laboratorio hemos demostrado previamente que Ang (1-7) tiene actividad anti-atrófica en modelos de atrofia muscular inducidos por angiotensina II y lipopolisacárido, dos modelos en los cuales hemos determinado además, alta expresión de MSTN muscular, sugiriendo que podria ser un factor común en la generación y/o mantención de la atrofia muscular en estos modelos. Además, se demostró recientemente que Ang (1-7) es capaz de disminuir la activación de la vía de señalización de Smad dependiente de TGF-β, una proteína perteneciente a la misma familia que MSTN. En conjunto, estos antecedentes permiten proponer en esta tesis la siguiente hipótesis: “Ang (1-7) disminuye la activación de la vía Smad y previene la inducción de parámetros de atrofia inducida por MSTN”. Para probar esta hipótesis, se utilizo un modelo in vitro de miotubos C2C12 los que fueron tratados con MSTN en presencia o ausencia de Ang (1-7) y se evaluó: 1) el efecto de Ang (1-7) sobre la vía de señalización de Smad activada por MSTN, determinada por la medición de los niveles proteicos de Smad2 y Smad3 fosforiladas; 2) el efecto de Ang (1-7) sobre parámetros de atrofia inducidos por MSTN, determinado por la medición del diámetro de miotubos, niveles proteicos y de mRNA de MHC, Atrogina-1 y MuRF-1. Los resultados indican que Ang (1-7) atenua la activación de la vía Smad (disminuye los niveles proteicos de fosfo-Smad2 y fosfo-Smad3) y tiene un efecto preventivo sobre parámetros de atrofia inducidos por MSTN (previene la disminución del diámetro de miotubos y niveles proteicos de MHC, asi como también el aumento en los niveles de mRNA y proteicos de Atrogina-1 y MuRF-1). En resumen, estos resultados sugieren que Ang (1-7) es un péptido capaz de disminuir la activación de vía de señalización de Smad y prevenir la inducción de parámetros de atrofia inducida por MSTN.
Role of the ubiquitin-proteasome system in the sarcopenic-like phenotype induced by CCL5/RANTES
(Page Press Publications, 2024-02) Conejeros-Lillo, Sabrina; Aguirre, Francisco; Cabrera, Daniel; Simon, Felipe; Peñailillo, Luis; Cabello-Verrugio, Claudio
Sarcopenia is characterized by reduced muscle strength and mass, and a decline in muscle fiber diameter and amount of sarcomeric proteins. Sarcopenia involves the activation of the ubiquitin-proteasome system (UPS). MuRF-1 and atrogin-1 are E3 ubiquitin ligases belonging to UPS, leading to proteolysis mediated by the PSMB 5, 6, and 7 subunits of 20S proteasome. CCL5/RANTES induces a sarcopenic-like effect in muscle cells. The present work explored the impact of CCL5 on UPS components and the influence of UPS on its sarcopenic-like effect. We demonstrated that CCL5 increased MuRF-1 and atrogin-1 protein levels and mRNA levels of subunits PSMB 5, 6, and 7. We used the MG132 inhibitor to elucidate the role of the 20S proteasome in the CCL5-induced sarcopenic-like effect. This inhibitor prevented the decrease in troponin and MHC protein levels and partially stopped the reduction in the diameter of single-isolated flexor digitorum brevis (FDB) muscle fibers induced by CCL5. These findings indicate that CCL5 actively modulates the UPS. Moreover, our results show the direct participation of UPS in the sarcopenic-like phenotype induced by CCL5

Examinando por Autor "Cabello-Verrugio, Claudio"

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