RESUMEN
Neuroinflammation and brain oxidative stress are recognized as significant contributors to hypertension including salt sensitive hypertension. Extracellular vesicles (EVs) play an essential role in intercellular communication in various situations, including physiological and pathological ones. Based on this evidence, we hypothesized that EVs derived from the brains of hypertensive rats with salt sensitivity could trigger neuroinflammation and oxidative stress during hypertension development. To test this hypothesis, we compared the impact of EVs isolated from the brains of hypertensive Dahl Salt-Sensitive rats (DSS) and normotensive Sprague Dawley (SD) rats on inflammatory factors and mitochondrial reactive oxygen species (mtROS) production in primary neuronal cultures and brain cardiovascular relevant regions, including the hypothalamic paraventricular nucleus (PVN) and lamina terminalis (LT). We found that brain-derived DSS-EVs significantly increased the mRNA levels of proinflammatory cytokines (PICs) and chemokines, including TNFα, IL1ß, CCL2, CCL5, and CCL12, as well as the transcriptional factor NF-κB in neuronal cultures. DSS-EVs also induced oxidative stress in neuronal cultures, as evidenced by elevated NADPH oxidase subunit CYBA coding gene mRNA levels and persistent mtROS elevation. When DSS-EVs were injected into the brains of normal SD rats, the mRNA levels of PICs, chemokines, and the chronic neuronal activity marker FOSL1 were significantly increased in the PVN and LT. Furthermore, DSS-EVs caused mtROS elevation in brain PVN and LT, particularly in neurons. Our study reveals a novel role for brain-derived EVs from hypertensive rats in triggering neuroinflammation, upregulating chemokine expression, and inducing excessive ROS production. These findings provide insight into the complex interactions between EVs and hypertension-associated processes, offering potential therapeutic targets for hypertension-linked neurological complications.
RESUMEN
Conducting in vivo brain imaging can be a challenging task due to the complexity of brain tissue and the strict requirements for safe and effective imaging agents. However, a new fluorescent dye called Cy5-PEG2 has been developed that selectively accumulates in mitochondria, enabling the visualization of these essential organelles in various cell lines. This dye is versatile and can be used for the real-time monitoring of mitochondrial dynamics in living cells. Moreover, it can cross the blood-brain barrier, making it a promising tool for noninvasive in vivo brain imaging. Based on the assessment of glial cell responses in the hippocampus and neocortex regions using GFAP and Iba1 biomarkers, Cy5-PEG2 seems to have minimal adverse effects on brain immune response or neuronal health. Therefore, this mitochondria-targeting fluorescent dye has the potential to advance our understanding of mitochondrial dynamics and function within the broader context of whole-brain physiology and disease progression. However, further research is needed to evaluate the safety and efficacy of Cy5-PEG2.
RESUMEN
Mitochondrial dysfunction is implicated in both brain tumors and neurodegenerative diseases, leading to various cellular abnormalities that can promote tumor growth and resistance to thera-pies, as well as impaired energy production and compromised neuronal function. Developing targeted therapies aimed at restoring mitochondrial function and improving overall cellular health could potentially be a promising approach to treating these conditions. Brain-derived exosomes (BR-EVs) have emerged as potential drug delivery vessels for neurological conditions. Herein, we report a new method for creating mitochondria-targeting exosomes and test its application in vitro and in vivo.
RESUMEN
Mitochondrial dysfunction is associated with various health conditions, including cardiovascular and neurodegenerative diseases. Mitochondrial-targeting therapy aims to restore or enhance mitochondrial function to treat or alleviate these conditions. Exosomes, small vesicles that cells secrete, containing a variety of biomolecules, are critical in cell-to-cell communication and have been studied as potential therapeutic agents. Exosome-based therapy has the potential to treat both cardiovascular and neurodegenerative diseases. Combining these two approaches involves using exosomes as carriers to transport mitochondrial-targeting agents to dysfunctional or damaged mitochondria within target cells. This article presents a new technique for engineering brain-derived exosomes that target mitochondria and has demonstrated promise in initial tests with primary neuron cells and healthy rats. This promising development represents a significant step forward in treating these debilitating conditions.
RESUMEN
Many organelles, such as lysosomes and mitochondria, maintain a pH that is different from the cytoplasmic pH. These pH differences have important functional ramifications for those organelles. Many cellular events depend upon a well-compartmentalized distribution of H+ ions spanning the membrane for the optimal function. Cells have developed a variety of mechanisms that enable the regulation of organelle pH. However, the measurement of organellar acidity/alkalinity in living cells has remained a challenge. Currently, most existing probes for the estimation of intracellular pH show a single -organelle targeting capacity. Such probes provide data that fails to comprehensively reveal the pathological and physiological roles and connections between mitochondria and lysosomes in different species. Mitochondrial and lysosomal functions are closely related and important for regulating cellular homeostasis. Accordingly, the design of a single fluorescent probe that can simultaneously target mitochondria and lysosomes is highly desirable, enabling a better understanding of the crosstalk between these organelles. We report the development of a novel fluorescent sensor, rhodamine-coumarin pH probe (RCPP), for detection of organellar acidity/alkalinity. RCPP simultaneously moves between mitochondrion and lysosome subcellular locations, facilitating the simultaneous monitoring of pH alterations in mitochondria and lysosomes.
RESUMEN
Clinically approved therapeutics that mitigate chemotherapy-induced cardiotoxicity, a serious adverse effect of chemotherapy, are lacking. The aim of this study was to determine the putative protective capacity of a novel indole alkaloid derivative B (IADB) against 5-fluorouracil (5-FU)-induced cardiotoxicity. To assess the free-radical scavenging activities of IADB, the acetylcholine-induced relaxation assay in rat thoracic aorta was used. Further, IADB was tested in normal and cancer cell lines with assays gauging autophagy induction. We further examined whether IADB could attenuate cardiotoxicity in 5-FU-treated male ICR mice. We found that IADB could serve as a novel bifunctional agent (displaying both antioxidant and autophagy-modulating activities). Further, we demonstrated that IADB induced production of cytosolic autophagy-associated structures in both cancer and normal cell lines. We observed that IADB cytotoxicity was much lower in normal versus cancer cell lines, suggesting an enhanced potency toward cancer cells. The cardiotoxicity induced by 5-FU was significantly relieved in animals pretreated with IADB. Taken together, IADB treatment, in combination with chemotherapy, may lead to reduced cardiotoxicity, as well as the reduction of anticancer drug dosages that may further improve chemotherapeutic efficacy with decreased off-target effects. Our data suggest that the use of IADB may be therapeutically beneficial in minimizing cardiotoxicity associated with high-dose chemotherapy. On the basis of the redox status difference between normal and tumor cells, IADB selectively induces autophagic cell death, mediated by reactive oxygen species overproduction, in cancer cells. This novel mechanism could reveal novel therapeutic targets in chemotherapy-induced cardiotoxicity.
RESUMEN
Mitochondrial oxidative damage contributes to a wide range of pathologies including ischemia/reperfusion injury. Accordingly, protecting mitochondria from oxidative damage should possess therapeutic relevance. In the present study, we have designed and synthesized a series of novel indole-TEMPO conjugates that manifested good anti-inflammatory properties in a murine model of xylene-induced ear edema. We have demonstrated that these compounds can protect cells from simulated ischemia/reperfusion (s-I/R)-induced reactive oxygen species (ROS) overproduction and mitochondrial dysfunction. Furthermore, we have demonstrated that indole-TEMPO conjugates can attenuate organ damage induced in rodents via intestinal I/R injury. We therefore propose that the pharmacological profile and mechanism of action of these indole-TEMPO conjugates involve convergent roles, including the ability to decrease free radical production via lipid peroxidation which couples to an associated decrease in ROS-mediated activation of the inflammatory process. We further hypothesize that the protective effects of indole-TEMPO conjugates partially reside in maintaining optimal mitochondrial function.
Asunto(s)
Antiinflamatorios no Esteroideos/uso terapéutico , Antioxidantes/uso terapéutico , Óxidos N-Cíclicos/uso terapéutico , Indoles/uso terapéutico , Mitocondrias/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Daño por Reperfusión/tratamiento farmacológico , Animales , Antiinflamatorios no Esteroideos/administración & dosificación , Antiinflamatorios no Esteroideos/síntesis química , Antioxidantes/administración & dosificación , Antioxidantes/química , Aspirina/farmacología , Óxidos N-Cíclicos/administración & dosificación , Óxidos N-Cíclicos/síntesis química , Citocromos c/metabolismo , Células Endoteliales de la Vena Umbilical Humana , Humanos , Indoles/administración & dosificación , Indoles/síntesis química , Indoles/farmacología , Intestino Delgado/irrigación sanguínea , Intestino Delgado/metabolismo , Intestino Delgado/patología , Peroxidación de Lípido/efectos de los fármacos , Masculino , Ratones Endogámicos ICR , Mitocondrias/metabolismo , Simulación de Dinámica Molecular , Infiltración Neutrófila/efectos de los fármacos , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
We describe several novel curcumin analogues that possess both anti-inflammatory antioxidant properties and thrombolytic activities. The therapeutic efficacy of these curcumin analogues was verified in a mouse ear edema model, a rat arterial thrombosis assay, a free radical scavenging assay performed in PC12 cells, and in both in vitro and in vivo ischemia/reperfusion models. Our findings suggest that their protective effects partially reside in maintenance of optimal mitochondrial function.
Asunto(s)
Antiinflamatorios/farmacología , Curcumina/análogos & derivados , Depuradores de Radicales Libres/farmacología , Mitocondrias/efectos de los fármacos , Animales , Antiinflamatorios/química , Antiinflamatorios/uso terapéutico , Curcumina/farmacología , Curcumina/uso terapéutico , Citocromos c/metabolismo , Modelos Animales de Enfermedad , Edema/prevención & control , Ensayo de Inmunoadsorción Enzimática , Depuradores de Radicales Libres/química , Depuradores de Radicales Libres/uso terapéutico , Células Endoteliales de la Vena Umbilical Humana , Interleucina-6/sangre , Ratones , Microscopía Fluorescente , Mitocondrias/metabolismo , Músculo Esquelético/metabolismo , Músculo Esquelético/patología , Estrés Oxidativo/efectos de los fármacos , Células PC12 , Teoría Cuántica , Ratas , Especies Reactivas de Oxígeno/metabolismo , Daño por Reperfusión/tratamiento farmacológico , Daño por Reperfusión/patología , Factor de Necrosis Tumoral alfa/sangreRESUMEN
Mitochondrial oxidative damage contributes to a wide range of pathologies, including ischemia/reperfusion (I/R) injury, cardiovascular disorders and neurodegenerative diseases. Accordingly, protecting mitochondria from oxidative damage should possess therapeutic relevance. In the present study, we have designed and synthesized a series of novel kyotorphin-nitroxide hybrid molecules, and examined their free radical scavenging activities, in addition to their anti-inflammatory and analgesic activities. We have further characterized these compounds in a simulated I/R cellular model. Our findings suggest that the protective effects of kyotorphin-nitroxides partially reside in maintaining optimal mitochondrial function.
Asunto(s)
Analgésicos/farmacología , Antiinflamatorios no Esteroideos/farmacología , Antioxidantes/farmacología , Endorfinas/farmacología , Óxidos de Nitrógeno/farmacología , Analgésicos/síntesis química , Analgésicos/química , Animales , Antiinflamatorios no Esteroideos/síntesis química , Antiinflamatorios no Esteroideos/química , Antioxidantes/síntesis química , Antioxidantes/química , Relación Dosis-Respuesta a Droga , Edema/inducido químicamente , Edema/tratamiento farmacológico , Endorfinas/química , Ratones , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Estructura Molecular , Óxidos de Nitrógeno/química , Dolor/tratamiento farmacológico , Dimensión del Dolor/efectos de los fármacos , Relación Estructura-Actividad , XilenosRESUMEN
Mitochondrial oxidative stress has been implicated in aging, neurodegenerative diseases, diabetes, stroke, ischemia/reperfusion injury, age-related macular degeneration (AMD) and cancer. Recently, we developed two new mitochondria-targeting fluorescent probes, MitoProbes I/II, which specifically localize in mitochondria and employed both in vivo and in vitro for detection of mitochondrial oxidative stress. Here, we report the design and synthesis of these agents, as well as their utility for real-time imaging of mitochondrial oxidative stress in cells.
Asunto(s)
Colorantes Fluorescentes/metabolismo , Microscopía Confocal/métodos , Mitocondrias/metabolismo , Células HeLa , Humanos , Modelos Moleculares , Estrés OxidativoRESUMEN
Intracellular pH plays an important role in the response to cancer invasion. We have designed and synthesized a series of new fluorescent probes (Superior LysoProbes) with the capacity to label acidic organelles and monitor lysosomal pH. Unlike commercially available fluorescent dyes, Superior LysoProbes are lysosome-specific and are highly stable. The use of Superior LysoProbes facilitates the direct visualization of the lysosomal response to lobaplatin elicited in human chloangiocarcinoma (CCA) RBE cells, using confocal laser scanning microscopy. Additionally, we have characterized the role of lysosomes in autophagy, the correlation between lysosome function and microtubule strength, and the alteration of lysosomal morphology during apoptosis. Our findings indicate that Superior LysoProbes offer numerous advantages over previous reagents to examine the intracellular activities of lysosomes.
Asunto(s)
Apoptosis , Colorantes Fluorescentes , Lisosomas/metabolismo , Imagen Molecular , Coloración y Etiquetado , Apoptosis/efectos de los fármacos , Transporte Biológico , Puntos de Control del Ciclo Celular , Línea Celular , Cloroquina/farmacología , Ciclobutanos/farmacología , Citoesqueleto/metabolismo , Colorantes Fluorescentes/química , Colorantes Fluorescentes/metabolismo , Humanos , Concentración de Iones de Hidrógeno , Espacio Intracelular/metabolismo , Microscopía Confocal , Compuestos Organoplatinos/farmacologíaRESUMEN
We report the design, synthesis and application of several new fluorescent probes (LysoProbes I-VI) that facilitate lysosomal pH monitoring and characterization of lysosome-dependent apoptosis. LysoProbes are superior to commercially available lysosome markers since the fluorescent signals are both stable and highly selective, and they will aid in characterization of lysosome morphology and trafficking. We predict that labeling of cancer cells and solid tumor tissues with LysoProbes will provide an important new tool for monitoring the role of lysosome trafficking in cancer invasion and metastasis.
Asunto(s)
Colorantes Fluorescentes/química , Lisosomas/química , Células HeLa , Humanos , Concentración de Iones de Hidrógeno , Lactosa/química , Lisosomas/fisiología , Espectrometría de Fluorescencia , Coloración y EtiquetadoRESUMEN
We synthesized and evaluated a series of acidic fluorescent pH probes exhibiting robust pH dependence, high sensitivity and photostability, and excellent cell membrane permeability. Titration analyses indicated that probe 3 could increase its fluorescence intensity 800-fold between pH 8.0 and 4.1. Additionally, its pK(a) value is optimal for intracellular probing of acidic organelles. Fluorescent imaging of HepG2 and Hela cells further revealed that probe 3 demonstrates outstanding capacity for monitoring of intracellular [H(+)] levels. The easily accessible terminal alkyne/azido function groups of these probes offer the possibility of rapidly constructing sensor molecule libraries using 'click' chemistry.
Asunto(s)
Técnicas Biosensibles , Colorantes Fluorescentes/síntesis química , Lactamas/síntesis química , Rodaminas/síntesis química , Ácidos , Permeabilidad de la Membrana Celular , Citoplasma/química , Colorantes Fluorescentes/química , Células HeLa , Células Hep G2 , Humanos , Concentración de Iones de Hidrógeno , Lactamas/química , Orgánulos/química , Protones , Rodaminas/química , Espectrometría de FluorescenciaRESUMEN
We have recently developed new Tempo-PEG-RGDs conjugates and have quantitatively examined their antithrombotic and antioxidant capabilities. These compounds were therapeutically beneficial when characterized in both in vitro platelet aggregation assays and a rat model of arterial thrombosis. Moreover, these compounds demonstrated significant protection from organ damage in a rat model of ischemia/reperfusion. Our data indicate that Tempo-PEG-RGDs represent a new class of adjuvants with therapeutic efficacy in acute and transient ischemic damage.
Asunto(s)
Óxidos N-Cíclicos/farmacología , Depuradores de Radicales Libres/síntesis química , Oligopéptidos/síntesis química , Estrés Oxidativo/fisiología , Polietilenglicoles/síntesis química , Daño por Reperfusión/tratamiento farmacológico , Animales , Depuradores de Radicales Libres/farmacología , Miembro Posterior/irrigación sanguínea , Histocitoquímica , Malondialdehído/sangre , Músculo Esquelético/fisiopatología , Oligopéptidos/farmacología , Agregación Plaquetaria/fisiología , Polietilenglicoles/farmacología , Ratas , Daño por Reperfusión/sangre , Superóxido Dismutasa/sangreRESUMEN
The synthesis, characteristics, and biological applications of a series of new rhodamine nitroxide fluorescent probes that enable imaging of hydroxyl radicals (â¢OH) in living cells are described. These probes are highly selective for â¢OH in aqueous solution, avoiding interference from other reactive oxygen species (ROS), and they facilitate â¢OH imaging in biologically active samples. The robust nature of these probes (high specificity and selectivity, and facile synthesis) offer distinct advantages over previous methods for â¢OH detection.
Asunto(s)
Colorantes Fluorescentes/análisis , Radical Hidroxilo/análisis , Espacio Intracelular/química , Óxidos de Nitrógeno/análisis , Rodaminas/química , Línea Celular , Supervivencia Celular , Colorantes Fluorescentes/síntesis química , Humanos , Radical Hidroxilo/química , Estructura Molecular , Óxidos de Nitrógeno/química , Estrés OxidativoRESUMEN
We have synthesized a series of new ß-carboline-tripeptide conjugates, and examined their anti-inflammatory properties in a mouse model of xylene-induced ear edema. The analgesic capacity of these compounds was further evaluated in a rodent tail flick assay. Our results indicate that ß-carboline conjugate 4a manifests potent anti-inflammatory and analgesic activity while exerting a protective effect against mesenteric ischemia/reperfusion (I/R) injury in the rat.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Carbolinas/química , Arterias Mesentéricas/efectos de los fármacos , Oligopéptidos/farmacología , Daño por Reperfusión/tratamiento farmacológico , Animales , Antiinflamatorios no Esteroideos/síntesis química , Antiinflamatorios no Esteroideos/química , Cristalografía por Rayos X , Relación Dosis-Respuesta a Droga , Edema/inducido químicamente , Edema/prevención & control , Masculino , Arterias Mesentéricas/patología , Ratones , Ratones Endogámicos ICR , Modelos Moleculares , Simulación de Dinámica Molecular , Estructura Molecular , Oligopéptidos/síntesis química , Oligopéptidos/química , Ratas , Ratas Wistar , Daño por Reperfusión/patología , Estereoisomerismo , Xilenos/antagonistas & inhibidoresRESUMEN
We describe a novel class of ß-carboline alkaloid-peptide conjugates that possess both free radical scavenging and thrombolytic activity. These conjugates demonstrate therapeutic efficacy in a rat arterial thrombosis assay, as well as free radical scavenging capacity as evaluated in a PC12 cell survival assay. Our results indicate that ß-carboline alkaloid-peptide conjugate 26a exerts a significant protective effect against local and remote organ injury induced by limb I/R injury in the rat.
Asunto(s)
Alcaloides/farmacología , Carbolinas/farmacología , Extremidades/irrigación sanguínea , Isquemia/tratamiento farmacológico , Péptidos/farmacología , Daño por Reperfusión/tratamiento farmacológico , Enfermedad Aguda , Alcaloides/síntesis química , Alcaloides/química , Animales , Carbolinas/síntesis química , Carbolinas/química , Supervivencia Celular/efectos de los fármacos , Extremidades/patología , Depuradores de Radicales Libres/metabolismo , Isquemia/patología , Simulación de Dinámica Molecular , Estructura Molecular , Células PC12 , Péptidos/síntesis química , Péptidos/química , Ratas , Daño por Reperfusión/patología , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
Synthetic oligodeoxynucleotide is purified by capping failure sequences with an acrylated phosphoramidite followed by polymerization and product extraction. The method is suitable for large scale oligonucleotide drug purification.
Asunto(s)
Oligodesoxirribonucleótidos/síntesis química , Secuencia de Bases , Cromatografía Líquida de Alta Presión , Oligodesoxirribonucleótidos/química , Oligodesoxirribonucleótidos/aislamiento & purificación , Compuestos Organofosforados/química , PolimerizacionRESUMEN
To develop more potent therapeutic agents with therapeutic efficacy for ischemia/reperfusion (I/R) injury, we linked an antiinflammatory moiety (1,3-dioxane derivative) to the key pharmacophoric moiety of melatonin. We hypothesized that the resulting new indole derivatives might induce a synergistic protection against oxidative damage associated with I/R injury. Our results indicate that one of these indole derivatives (7) manifests potent antiinflammatory antioxidant effects and exerts a protective effect against skeletal muscle injury and associated lung injury following limb I/R in rats.
Asunto(s)
Antiinflamatorios no Esteroideos/síntesis química , Antioxidantes/síntesis química , Indoles/síntesis química , Daño por Reperfusión/tratamiento farmacológico , Animales , Antiinflamatorios no Esteroideos/química , Antiinflamatorios no Esteroideos/farmacología , Antioxidantes/química , Antioxidantes/farmacología , Permeabilidad Capilar , Depuradores de Radicales Libres/síntesis química , Depuradores de Radicales Libres/química , Depuradores de Radicales Libres/farmacología , Miembro Posterior/irrigación sanguínea , Indoles/química , Indoles/farmacología , Inflamación/tratamiento farmacológico , Peroxidación de Lípido/efectos de los fármacos , Lesión Pulmonar/tratamiento farmacológico , Lesión Pulmonar/metabolismo , Lesión Pulmonar/patología , Ratones , Músculo Esquelético/irrigación sanguínea , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/patología , Células PC12 , Ratas , Daño por Reperfusión/metabolismo , Daño por Reperfusión/patología , Relación Estructura-ActividadRESUMEN
Renal ischemia/reperfusion is a common cause of acute renal failure. Glycine is an effective anti-inflammatory, cytoprotective agent and is reported to have a beneficial effect against ischemia/reperfusion injury in various organs. Previous research notes that free radicals and inflammatory leukocytes both play important roles in the pathogenesis of renal ischemia/reperfusion injury. To develop new therapeutic agents against renal ischemia/reperfusion injury, we sought to link an antioxidant moiety (nitronyl nitroxide) to glycine in the hope that the resulting glycine-nitronyl nitroxide conjugate (GNN) would provide a synergetic protection against renal ischemia/reperfusion injury. In this manuscript, we report the synthesis and biological evaluation of the GNN conjugate. The biological activity of the GNN conjugate was evaluated in an in vivo rat model of renal ischemia/reperfusion induced injury and oxidative change. Since the GNN conjugate markedly reduced elevated levels of tissue lipid peroxidation and attenuated renal dysfunction in rats subjected to renal ischemia/reperfusion, it might be possible to develop the GNN conjugate into a potential therapeutic agent against renal ischemia/reperfusion injury.