RESUMEN
BACKGROUND: This study explores the potential role of Thioredoxin-interacting protein (TXNIP) silencing in endothelial colony-forming cells (ECFCs) within the scope of age-related comorbidities and impaired vascular repair. We aim to elucidate the effects of TXNIP silencing on vasculogenic properties, paracrine secretion, and neutrophil recruitment under conditions of metabolic stress. METHODS: ECFCs, isolated from human blood cord, were transfected with TXNIP siRNA and exposed to a high glucose and ß-hydroxybutyrate (BHB) medium to simulate metabolic stress. We evaluated the effects of TXNIP silencing on ECFCs' functional and secretory responses under these conditions. Assessments included analyses of gene and protein expression profiles, vasculogenic properties, cytokine secretion and neutrophil recruitment both in vitro and in vivo. The in vivo effects were examined using a murine model of hindlimb ischemia to observe the physiological relevance of TXNIP modulation under metabolic disorders. RESULTS: TXNIP silencing did not mitigate the adverse effects on cell recruitment, vasculogenic properties, or senescence induced by metabolic stress in ECFCs. However, it significantly reduced IL-8 secretion and consequent neutrophil recruitment under these conditions. In a mouse model of hindlimb ischemia, endothelial deletion of TXNIP reduced MIP-2 secretion and prevented increased neutrophil recruitment induced by age-related comorbidities. CONCLUSIONS: Our findings suggest that targeting TXNIP in ECFCs may alleviate ischemic complications exacerbated by metabolic stress, offering potential clinical benefits for patients suffering from age-related comorbidities.
Asunto(s)
Proteínas Portadoras , Interleucina-8 , Infiltración Neutrófila , Estrés Fisiológico , Animales , Interleucina-8/metabolismo , Interleucina-8/genética , Proteínas Portadoras/metabolismo , Proteínas Portadoras/genética , Humanos , Ratones , Infiltración Neutrófila/efectos de los fármacos , Células Progenitoras Endoteliales/metabolismo , Células Progenitoras Endoteliales/efectos de los fármacos , Isquemia/metabolismo , Isquemia/patología , ARN Interferente Pequeño/metabolismo , Tiorredoxinas/metabolismo , Tiorredoxinas/genética , Miembro Posterior/irrigación sanguínea , Ratones Endogámicos C57BL , Glucosa/metabolismoRESUMEN
Thioredoxin interacting protein (TXNIP) is a metabolism- oxidative- and inflammation-related marker induced in cardiovascular diseases and is believed to represent a possible link between metabolism and cellular redox status. TXNIP is a potential biomarker in cardiovascular and ischemic diseases but also a novel identified target for preventive and curative medicine. The goal of this review is to focus on the novelties concerning TXNIP. After an overview in TXNIP involvement in oxidative stress, inflammation and metabolism, the remainder of this review presents the clues used to define TXNIP as a new marker at the genetic, blood, or ischemic site level in the context of cardiovascular and ischemic diseases.
Asunto(s)
Biomarcadores/análisis , Enfermedades Cardiovasculares/patología , Proteínas Portadoras/metabolismo , Isquemia/patología , Terapia Molecular Dirigida , Tiorredoxinas/metabolismo , Animales , Enfermedades Cardiovasculares/metabolismo , Enfermedades Cardiovasculares/terapia , Humanos , Inflamación , Isquemia/metabolismo , Isquemia/terapia , Estrés OxidativoRESUMEN
Targeted therapies such as cabozantinib (CABO), pazopanib (PAZO), sorafenib (SORA), sunitinib (SUNI) and its main active metabolite N-desethyl-sunitinib (DST-SUNI), olaparib (OLA) and palbociclib (PALBO) display large pharmacokinetics variability impacting their responses in terms of efficacy or toxicity. For the monitoring of these drugs, an analytical method allowing to routinely measure their concentrations in human plasma is needed. Such a method has been developed and validated and is presented here. The chromatographic separation is achieved on a Zorbax Bonus-RP analytical column using an isocratic elution of 92% V/V of acetonitrile and 8% of water in 0.1% formic acid at a flow rate of 500⯵l/min for 0.5â¯min and then 300⯵l/min for 2â¯min. After a liquid-liquid extraction of plasma samples, a step of filtration is performed. This method was validated based on the EMA and French committee of accreditation guidelines. The analysis time is 2.5â¯min per run, and all analytes eluted within 0.53-1.61â¯min. The standard curves are linear over the range from 1 to 380â¯ng/ml for SUNI; from 4.3 to 450â¯ng/ml for DST-SUNI; from 6 to 1000â¯ng/ml for PALBO; from 75 to 5000â¯ng/ml for CABO, from 0.17 to 20⯵g/ml for OLA; from 0.35 to 40⯵g/ml for SORA and from 1.7 to 200⯵g/ml for PAZO. The method also showed satisfactory results in terms precision (below 9.5% for within-run and below 13% for between-run) and accuracy (below 13.5% for within-run and below 14% for between-run). After sampling, all the compounds are stable in whole blood at ambient temperature at least for 6â¯h and plasma are stable for 48â¯h at ambient temperature or 4⯰C. The method presented here allows to measure the concentrations of 7 targeted therapies in a routine setting. We moreover present here a method that is, to our knowledge, one of the first detailed method aimed at the measurement of palbociclib in human plasma in a routine setting, together with data useful for the management of samples in routine hospital practice.
Asunto(s)
Antineoplásicos , Cromatografía Líquida de Alta Presión/métodos , Compuestos Heterocíclicos con 1 Anillo , Antineoplásicos/sangre , Antineoplásicos/química , Antineoplásicos/metabolismo , Estabilidad de Medicamentos , Compuestos Heterocíclicos con 1 Anillo/sangre , Compuestos Heterocíclicos con 1 Anillo/química , Compuestos Heterocíclicos con 1 Anillo/metabolismo , Humanos , Límite de Detección , Modelos Lineales , Reproducibilidad de los Resultados , Espectrometría de Masas en Tándem/métodosRESUMEN
The inhibition of arginase is of substantial interest for the treatment of various diseases of public health interest including cardiovascular diseases. Using an ex vivo experiment on rat aortic rings and an in vitro assay with liver bovine purified arginase, it was demonstrated that several polyphenolic extracts from Cyperus and Carex species possess vasorelaxant properties and mammalian arginase inhibitory capacities. Phytochemical studies performed on these species led to the identification of eight compounds, including monomers, dimers, trimers, and tetramers of resveratrol. The potential of these stilbenes as inhibitors of mammalian arginase was assessed. Five compounds, scirpusin B (5), ε-viniferin (4), cyperusphenol B (6), carexinol A (7), and the new compound virgatanol (1), showed significant inhibition of arginase, with percentage inhibition ranging from 70% to 95% at 100 µg/mL and IC50 values between 12.2 and 182.1 µM, confirming that these stilbenes may be useful for the development of new pharmaceutical products.