RESUMEN
BACKGROUND: Alzheimer's disease (AD) is a progressive neurodegenerative disorder with poorly understood pathology. Elevated tau, phospho-tau and mitochondrial dysfunction are significantly correlated with an increased risk of AD and are therefore targets for disease-modifying therapy. In this study, we examined the effects of polyphenolic extracts from six different varieties of sorghum: Shawaya short black-1 (Black), IS1311C (Brown), QL33/QL36 (Red), B923296 (Red), QL12 (White), and QL33 (Red) on the attenuation of beta amyloid-induced phospho-tau levels, total tau levels, and mitochondrial dysfunction in neuronal cells. METHOD: Tau proteins (231 (pT231), Serine- 199 (pS199), and total tau proteins (T-tau)) were detected and quantified using sandwich ELISA kits, while mitochondrial dysfunction was measured in terms of mitochondrial membrane potential (Δψm) and adenosine triphosphate (ATP) levels. RESULTS: Almost all varieties of the sorghum extracts reduced the beta amyloid-induced pS199 and pT231 levels (p ≤ 0.05). The optimum concentration of QL33/QL36 (1000 µg/mL), QL12 (2000 µg/mL), and QL33 (2000 µg/mL) strongly attenuated the phospho-tau level. Sorghum IS1311C (750 µg/mL) showed the highest Δψm reduction (39.8%), whereas QL33 (2000 µg/mL) most strongly improved the ATP level (37.7%) (p ≤ 0.01). For both Δψm and ATP assays, the least activity was observed in sorghum B923296 at 21% and 25.5%, respectively (p ≤ 0.01). CONCLUSIONS: The polyphenol extracts from sorghum attenuated the tau toxicity and Aß-induced mitochondrial dysfunction in a variety- and dose-dependent manner and made a promising disease-modifying agent against AD. However, extensive research is needed to validate the efficacy of the sorghum extracts prior to animal and clinical studies.
Asunto(s)
Enfermedad de Alzheimer , Potencial de la Membrana Mitocondrial , Mitocondrias , Extractos Vegetales , Sorghum , Proteínas tau , Sorghum/química , Proteínas tau/metabolismo , Enfermedad de Alzheimer/metabolismo , Enfermedad de Alzheimer/tratamiento farmacológico , Fosforilación , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Extractos Vegetales/farmacología , Humanos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Péptidos beta-Amiloides/metabolismo , Adenosina Trifosfato/metabolismo , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Polifenoles/farmacologíaRESUMEN
Accurately and sensitively identifying and killing cancer cells, especially those in deep tissues, is of paramount importance but presents significant challenges. Herein, a membrane protein and adenosine triphosphate (ATP)-driven DNA logic gate-modified liposome is designed to coat zinc peroxide (ZP) nanoparticles integrated with nanozymes (ZP/RuTe@L/DNA) to accurately identify and induce cell apoptosis in cancer cells through a reactive oxygen species (ROS)-mediated mechanism under acid conditions in cancer cells. In this system, DNA logic gate-functionalized liposomes are loaded with ZP and nanozymes, while HeLa cancer cells are functionalized with a DNA segment that is complementary to a segment of the DNA logic gate. For the DNA logic gate, a DNA aptamer was employed for membrane protein recognition, and another aptamer was used for the response of extracellular ATP. Activation of the DNA logic gate occurs only when both biomarkers are simultaneously present. Once activated, the DNA logic gate-modified liposome could hybridize with DNA segment-modified HeLa cells, leading to liposome-HeLa cell fusion and the release of ZP/RuTe into HeLa cells. Under acid conditions, ZP could decompose to release H2O2 and Zn2+, which could promote the production of â¢O2- and H2O2 by inhibiting the electron transport chain. Concurrently, the released RuTe exhibits glutathione (GSH) depletion and peroxidase (POD) and nicotinamide adenine dinucleotide (NADH) peroxidase-like activities, generating highly toxic hydroxyl radical (â¢OH), disrupting the cellular redox homeostasis, and inducing cell apoptosis. The ZP/RuTe@L/DNA system could not only accurately detect cancer cells in complex cell mixtures but also present a novel method for liposome-membrane fusion processes in drug delivery. This study presents significant potential for application in precise cancer diagnosis and therapy.
Asunto(s)
Liposomas , Humanos , Células HeLa , Liposomas/química , Apoptosis/efectos de los fármacos , Fusión de Membrana/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , ADN/química , ADN/metabolismo , Peróxido de Hidrógeno/farmacología , Aptámeros de Nucleótidos/química , Aptámeros de Nucleótidos/farmacología , Adenosina Trifosfato/metabolismo , Adenosina Trifosfato/química , Neoplasias/tratamiento farmacológico , Neoplasias/metabolismo , Neoplasias/patología , Antineoplásicos/farmacología , Antineoplásicos/químicaRESUMEN
Age-related macular degeneration (AMD) is a major cause of vision loss among adults. We investigated the protective effects of passion fruit seed extract (PFSE) and its rich polyphenol piceatannol in an AMD cell model in which human retinal pigment epithelial ARPE-19 cells were exposed to hydrogen peroxide (H2O2). Using a cell viability WST-8 assay, we revealed that PFSE and piceatannol increased the cellular viability of ARPE-19 cells by 130% and 133%, respectively. Moreover, PFSE and piceatannol recovered the cell viability of ARPE-19 cells, which had decreased to 60% owing to H2O2-induced damage, to approximately 84% and 89%, respectively. In addition, we found that the treatment of ARPE-19 cells with H2O2 decreased the mitochondrial and glycolytic ATP production rate to approximately 54% that of healthy control ARPE-19 cells using a Seahorse extracellular flux analyzer. Furthermore, pretreatment with PFSE and piceatannol restored the oxidative stress-induced decrease in the mitochondrial and glycolytic ATP production rate to approximately 97% and 82%, respectively. These results indicated the cytoprotective effects of PFSE and piceatannol against oxidative stress in human ARPE-19 cells by resolving the dysfunction of mitochondrial and glycolytic energy metabolism.
Asunto(s)
Supervivencia Celular , Peróxido de Hidrógeno , Estrés Oxidativo , Passiflora , Extractos Vegetales , Epitelio Pigmentado de la Retina , Semillas , Humanos , Epitelio Pigmentado de la Retina/efectos de los fármacos , Epitelio Pigmentado de la Retina/metabolismo , Epitelio Pigmentado de la Retina/citología , Peróxido de Hidrógeno/toxicidad , Extractos Vegetales/farmacología , Semillas/química , Línea Celular , Passiflora/química , Supervivencia Celular/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Adenosina Trifosfato/metabolismo , Estilbenos/farmacología , Glucólisis/efectos de los fármacos , Mitocondrias/metabolismo , Mitocondrias/efectos de los fármacos , Degeneración Macular/metabolismo , Degeneración Macular/patología , Degeneración Macular/tratamiento farmacológicoRESUMEN
Sublethal tumor cells have an urgent need for energy, making it common for them to switch metabolic phenotypes between glycolysis and oxidative phosphorylation (OXPHOS) for compensatory energy supply; thus, the synchronous interference of dual metabolic pathways for limiting energy level is essential in inhibiting sublethal tumor growth. Herein, a multifunctional nanoplatform of Co-MOF-loaded anethole trithione (ADT) and myristyl alcohol (MA), modified with GOx and hyaluronic acid (HA) was developed, namely, CAMGH. It could synchronously interfere with dual metabolic pathways including glycolysis and OXPHOS to restrict the adenosine triphosphate (ATP) supply, achieving the inhibition to sublethal tumors after microwave (MW) thermal therapy. Under low-power MW irradiation, CAMGH induced certain tumor thermal damage while ensuring the safety of the surrounding normal tissues. The loaded GOx consumed glucose in tumors, undoubtedly blocking the main energy supply pathway, the glycolytic pathway. Then, H2O2 generated from GOx reacted with Co2+ to produce cytotoxic ·OH, combining with the released H2S from ADT to co-obstruct OXPHOS and then synergy with the above glycolysis blocking for a more effective ATP inhibition. The powerful depletion of ATP caused significant suppression of damage resistance protein upregulated after thermal stimulation, i.e., HSP90, and then the activation of caspase-3, achieving the simultaneous reversal of heat resistance and apoptosis resistance. Altogether, the CAMGH-based synchronous interference of dual metabolic pathways shows the potential to break down tumor self-repair, presenting an alternative strategy to enhance the therapeutic effect of tumor MW thermal therapy.
Asunto(s)
Microondas , Animales , Ratones , Humanos , Sulfuro de Hidrógeno/química , Sulfuro de Hidrógeno/metabolismo , Sulfuro de Hidrógeno/farmacología , Línea Celular Tumoral , Fosforilación Oxidativa/efectos de los fármacos , Fosforilación Oxidativa/efectos de la radiación , Neoplasias/metabolismo , Neoplasias/patología , Neoplasias/tratamiento farmacológico , Neoplasias/terapia , Glucólisis/efectos de los fármacos , Hipertermia Inducida , Adenosina Trifosfato/metabolismo , Adenosina Trifosfato/química , Ratones Endogámicos BALB C , Antineoplásicos/farmacología , Antineoplásicos/química , Redes y Vías Metabólicas/efectos de los fármacosRESUMEN
Neurite degeneration (ND) precedes cell death in many neurodegenerative diseases. However, it remains unclear how this compartmentalized cell death process is orchestrated in the central nervous system (CNS). The establishment of a CNS axotomy model (using modified 3D LUHMES cultures) allowed us to study metabolic control of ND in human midbrain-derived neurons without the use of toxicants or other direct disturbance of cellular metabolism. Axotomy lead to a loss of the NAD+ synthesis enzyme NMNAT2 within 2 h and a depletion of NAD+ within 4-6 h. This process appeared specific, as isolated neurites maintained ATP levels and a coupled mitochondrial respiration for at least 6 h. In the peripheral nervous system (PNS) many studies observed that NAD+ metabolism, in particular by the NADase SARM1, plays a major role in the ND occurring after axotomy. Since neither ferroptosis nor necroptosis, nor caspase-dependent apoptosis seemed to be involved in neurite loss, we investigated SARM1 as potential executioner (or controller). Knock-down or expression of a dominant-negative isoform of SARM1 indeed drastically delayed ND. Various modifications of NAD+ metabolism known to modulate SARM1 activity showed the corresponding effects on ND. Moreover, supplementation with NAD+ attenuated ND. As a third approach to investigate the role of altered NAD+ metabolism, we made use of the WLD(s) protein, which has been found in a mutant mouse to inhibit Wallerian degeneration of axons. This protein, which has a stable NMNAT activity, and thus can buffer the loss of NMNAT2, protected the neurites by stabilizing neurite NAD+ levels. Thus CNS-type ND was tightly linked to neurite metabolism in multiple experimental setups. Based on this knowledge, several new strategies for treating neurodegenerative diseases can be envisaged.
Asunto(s)
NAD , Neuritas , Nicotinamida-Nucleótido Adenililtransferasa , Humanos , NAD/metabolismo , Neuritas/metabolismo , Nicotinamida-Nucleótido Adenililtransferasa/metabolismo , Nicotinamida-Nucleótido Adenililtransferasa/genética , Proteínas del Dominio Armadillo/metabolismo , Proteínas del Dominio Armadillo/genética , Animales , Sistema Nervioso Central/metabolismo , Sistema Nervioso Central/patología , Axotomía , Apoptosis , Neuronas/metabolismo , Neuronas/patología , Ratones , Mitocondrias/metabolismo , Células Cultivadas , Adenosina Trifosfato/metabolismoRESUMEN
Haemorrhoids are a common anorectal disease primarily treated through surgery, often leading to complications such as pain. The efficacy of acupuncture in relieving postoperative pain in mixed haemorrhoids has not been well-documented. This study included 90 patients undergoing haemorrhoid surgery and their Visual Analogue Score (VAS), inflammatory factor levels, Hamilton Depression Rating Scale (HAMD), and analgesic drug use were accessed. A rat incisional pain model was also constructed to monitor behavioral responses, with assessments including Sucrose Preference Test (SPT) and Open Field Test (OFT). The levels of ATP and proinflammatory cytokines in the dorsal root ganglion (DRG) were measured using luciferase assay and ELISA. We also examined P2×7 and ERK1/2 levels in DRG tissues of anal incisional pain rat model. In a Chronic Constriction Injury (CCI) rat model treated with BzATP, a potent agonist for P2×7 receptors, followed by acupuncture for 15 days, postoperative pain and behavioral responses were observed and assessed, alongside mechanistic studies of ATP and inflammatory factors in DRG tissues. Patients receiving acupuncture had significantly lower VAS scores, reduced levels of inflammatory factors, improved depression scores, and decreased analgesic drug use. In the animal model, acupuncture increased pain thresholds, improved behavioral responses, reduced ATP content and inflammatory factors, and modulated the P2×7/ERK axis. In the CCI model, BzATP increased P2×7 and ERK1/2 levels, pain sensitivity, and anxiety, which were mitigated by acupuncture. Our data suggest that acupuncture significantly alleviates postoperative pain following haemorrhoid surgery and modulates the pain response through the P2×7/ERK axis.
Asunto(s)
Terapia por Acupuntura , Ganglios Espinales , Hemorroides , Dolor Postoperatorio , Ratas Sprague-Dawley , Receptores Purinérgicos P2X7 , Animales , Dolor Postoperatorio/metabolismo , Dolor Postoperatorio/terapia , Dolor Postoperatorio/tratamiento farmacológico , Masculino , Ganglios Espinales/metabolismo , Ratas , Receptores Purinérgicos P2X7/metabolismo , Terapia por Acupuntura/métodos , Hemorroides/cirugía , Humanos , Persona de Mediana Edad , Adulto , Femenino , Sistema de Señalización de MAP Quinasas/fisiología , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Modelos Animales de Enfermedad , Adenosina Trifosfato/metabolismo , Dimensión del DolorRESUMEN
Purpose: To investigate the effect of Rho-associated protein kinase (ROCK) inhibitor Y27632 on bioenergetic capacity and resilience of corneal endothelial cells (CECs) under metabolic stress. Methods: Bovine CECs (BCECs) were treated with Y27632 and subjected to bioenergetic profiling using the Seahorse XFp Analyzer. The effects on adenosine triphosphate (ATP) production through oxidative phosphorylation and glycolysis were measured. BCECs were also challenged with monensin to induce metabolic stress. Cell viability, apoptosis, intracellular sodium levels, and hexokinase localization were assessed using calcein AM assay, flow cytometry, fluorescence imaging, and immunostaining, respectively. Results: Y27632 increased maximal ATP production rates via both oxidative phosphorylation and glycolysis, thereby expanding the overall bioenergetic capacity in BCECs. Under monensin-induced metabolic stress, ROCK inhibitor pretreatment significantly enhanced glycolytic ATP production and reduced apoptosis compared with untreated cells. Y27632 also facilitated sodium export by increasing Na/K-ATPase activity, as evidenced by lower intracellular sodium levels. Additionally, Y27632 promoted the translocation of hexokinase 2 to mitochondria under stress conditions, thereby enhancing glycolytic capacity. The effect of Y27632 on cell viability and sodium export was abrogated when cells were forced to rely on oxidative phosphorylation in galactose media, indicating that the protective effects of Y27632 are dependent on glycolytic ATP production under monensin stress. Conclusions: ROCK inhibitor Y27632 enhances the bioenergetic capacity of BCECs, allowing the cells to better withstand metabolic stress by rapidly generating ATP to meet increased energy demands, maintaining ion homeostasis and reducing apoptosis.
Asunto(s)
Adenosina Trifosfato , Amidas , Apoptosis , Supervivencia Celular , Endotelio Corneal , Metabolismo Energético , Piridinas , Quinasas Asociadas a rho , Animales , Endotelio Corneal/efectos de los fármacos , Endotelio Corneal/metabolismo , Bovinos , Quinasas Asociadas a rho/antagonistas & inhibidores , Quinasas Asociadas a rho/metabolismo , Metabolismo Energético/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Adenosina Trifosfato/metabolismo , Piridinas/farmacología , Apoptosis/efectos de los fármacos , Amidas/farmacología , Estrés Fisiológico/efectos de los fármacos , Citometría de Flujo , Glucólisis/efectos de los fármacos , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , ATPasa Intercambiadora de Sodio-Potasio/antagonistas & inhibidores , Fosforilación Oxidativa/efectos de los fármacosRESUMEN
Viruses are dependent on cellular energy metabolism for their replication, and the drug nitazoxanide (Alinia) was shown to interfere with both processes. Nitazoxanide is an uncoupler of mitochondrial oxidative phosphorylation (OXPHOS). Our hypothesis was that mitochondrial uncoupling underlies the antiviral effects of nitazoxanide. Tizoxanide (the active metabolite of nitazoxanide), its derivative RM4848 and the uncoupler CCCP were applied to a virus-releasing cell line to obtain the same increasing levels of mitochondrial uncoupling, hence identical impact on OXPHOS. A decrease in infectious viral particle release was observed and reflected the intensity of impact on OXPHOS, irrespective of the nature of the drug. The antiviral effect was significant although the impact on OXPHOS was modest (≤ 25%), and disappeared when a high concentration (25 mM) of glucose was used to enhance glycolytic generation of ATP. Accordingly, the most likely explanation is that moderate interference with mitochondrial OXPHOS induced rearrangement of ATP use and acquisition of infective properties of the viral particles be highly sensitive to this rearrangement. The antiviral effect of nitazoxanide has been supported by clinical trials, and nitazoxanide is considered a safe drug. However, serious adverse effects of the uncoupler dinitrophenol occurred when used to increase significantly metabolic rate with the purpose of weight loss. Taken together, while impairment of mitochondrial bioenergetics is an unwanted drug effect, moderate interference should be considered as a basis for therapeutic efficacy.
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Antivirales , Metabolismo Energético , Mitocondrias , Nitrocompuestos , Fosforilación Oxidativa , Tiazoles , Nitrocompuestos/farmacología , Tiazoles/farmacología , Fosforilación Oxidativa/efectos de los fármacos , Metabolismo Energético/efectos de los fármacos , Mitocondrias/metabolismo , Mitocondrias/efectos de los fármacos , Antivirales/farmacología , Humanos , Animales , Línea Celular , Replicación Viral/efectos de los fármacos , Adenosina Trifosfato/metabolismoRESUMEN
Redox balance is essential for sustaining normal physiological metabolic activities of life. In this study, we present a photocatalytic system to perturb the balance of NADH/NAD+ in oxygen-free conditions, achieving photocatalytic therapy to cure anaerobic bacterial infected periodontitis. Under light irradiation, the catalyst TBSMSPy+ can bind bacterial DNA and initiate the generation of radical species through a multi-step electron transfer process. It catalyzes the conversion from NADH to NAD+ (the turnover frequency up to 60.7 min-1), inhibits ATP synthesis, disrupts the energy supply required for DNA replication, and successfully accomplishes photocatalytic sterilization in an oxygen-free environment. The catalyst participates in the redox reaction, interfering with the balance of NADH/NAD+ contents under irradiation, so we termed this action as photoinduced redox imbalance. Additionally, animal experiments in male rats also validate that the TBSMSPy+ could effectively catalyze the NADH oxidation, suppress metabolism and stimulate osteogenesis. Our research substantiates the concept of photoinduced redox imbalance and the application of photocatalytic therapy, further advocating the development of such catalyst based on photoinduced redox imbalance strategy for oxygen-free phototherapy.
Asunto(s)
NAD , Oxidación-Reducción , Animales , NAD/metabolismo , Masculino , Ratas , Catálisis/efectos de la radiación , Luz , Fototerapia/métodos , Ratas Sprague-Dawley , Adenosina Trifosfato/metabolismoRESUMEN
We have shown that virus-specific CD4 and CD8 memory T cells (TM) induce autophagy after T cell receptor (TCR) engagement to provide free glutamine and fatty acids, including in people living with HIV-1 (PLWH). These nutrients fuel mitochondrial ATP generation through glutaminolysis and fatty acid oxidation (FAO) pathways, to fulfill the bioenergetic demands for optimal IL-21 and cytotoxic molecule production in CD4 and CD8 cells, respectively. Here, we expand our knowledge on how the metabolic events that occur in the mitochondria of virus-specific TM down-stream of the autophagy are regulated. We show that HSP60 chaperone positively regulates the protein levels for multiple glutaminolysis- and FAO-related enzymes, thereby actively fueling the levels of cellular alpha-ketoglutarate (αKG) and related mitochondrial ATP-dependent antiviral T cell immunity in both CD4 and CD8 TM. Finally, we provide a way to rescue defective ATP generation in mitochondria and dependent effector functions in virus-specific TM including anti-HIV-1 protective responses, when HSP60 expression is impaired after TCR engagement in patients, in the form of dimethyl 2-oxoglutarate (DMKG) supplementation.
Asunto(s)
Adenosina Trifosfato , Linfocitos T CD4-Positivos , Linfocitos T CD8-positivos , Chaperonina 60 , Interleucinas , Mitocondrias , Adenosina Trifosfato/metabolismo , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Mitocondrias/metabolismo , Mitocondrias/inmunología , Humanos , Chaperonina 60/metabolismo , Chaperonina 60/inmunología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , Interleucinas/metabolismo , Células T de Memoria/inmunología , Células T de Memoria/metabolismo , VIH-1/inmunología , Infecciones por VIH/inmunología , Infecciones por VIH/virología , Infecciones por VIH/metabolismo , Masculino , Autofagia , Proteínas MitocondrialesRESUMEN
BACKGROUND: Breast cancer, one of the most common forms of cancer, is associated with the highest cancer-related mortality among women worldwide. In comparison to other types of breast cancer, patients diagnosed with the triple-negative breast cancer (TNBC) subtype have the worst outcome because current therapies do not produce long-lasting responses. Hence, innovative therapies that produce persisting responses are a critical need. We previously discovered that hyperactivating purinergic receptors (P2RXs) by increasing extracellular adenosine triphosphate (eATP) concentrations enhances TNBC cell lines' response to chemotherapy. Heparan sulfate inhibits multiple extracellular ATPases, so it is a molecule of interest in this regard. In turn, heparanase degrades polysulfated polysaccharide heparan sulfate. Importantly, previous work suggests that breast cancer and other cancers express heparanase at high levels. Hence, as heparan sulfate can inhibit extracellular ATPases to facilitate eATP accumulation, it may intensify responses to chemotherapy. We postulated that heparanase inhibitors would exacerbate chemotherapy-induced decreases in TNBC cell viability by increasing heparan sulfate in the cellular microenvironment and hence, augmenting eATP. METHODS: We treated TNBC cell lines MDA-MB 231, Hs 578t, and MDA-MB 468 and non-tumorigenic immortal mammary epithelial MCF-10A cells with paclitaxel (cytotoxic chemotherapeutic) with or without the heparanase inhibitor OGT 2115 and/or supplemental heparan sulfate. We evaluated cell viability and the release of eATP. Also, we compared the expression of heparanase protein in cell lines and tissues by immunoblot and immunohistochemistry, respectively. In addition, we examined breast-cancer-initiating cell populations using tumorsphere formation efficiency assays on treated cells. RESULTS: We found that combining heparanase inhibitor OGT 2115 with chemotherapy decreased TNBC cell viability and tumorsphere formation through increases in eATP and activation of purinergic receptors as compared to TNBC cells treated with single-agent paclitaxel. CONCLUSION: Our data shows that by preventing heparan sulfate breakdown, heparanase inhibitors make TNBC cells more susceptible to chemotherapy by enhancing eATP concentrations.
Asunto(s)
Adenosina Trifosfato , Supervivencia Celular , Glucuronidasa , Heparitina Sulfato , Neoplasias de la Mama Triple Negativas , Humanos , Heparitina Sulfato/metabolismo , Neoplasias de la Mama Triple Negativas/tratamiento farmacológico , Neoplasias de la Mama Triple Negativas/metabolismo , Neoplasias de la Mama Triple Negativas/patología , Femenino , Línea Celular Tumoral , Glucuronidasa/metabolismo , Adenosina Trifosfato/metabolismo , Supervivencia Celular/efectos de los fármacos , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Paclitaxel/farmacología , Paclitaxel/uso terapéutico , Microambiente Tumoral/efectos de los fármacosRESUMEN
This study evaluates the safety and potential benefits of PBM on pancreatic beta cells and islets. PBM was applied to insulin-secreting cell lines (MIN6) and rat pancreatic islets using a 670 nm light source, continuous output, with a power density of 2.8 mW/cm², from 5 s to several 24 h. Measure of cell viability, insulin secretion, mitochondrial function, ATP content, and cellular respiration were assessed. Additionally, a diabetic rat model is used for islet transplantation (pre-conditioning with PBM or not) experiments. Short and long-term PBM exposure did not affect beta cell islets viability, insulin secretion nor ATP content. While short-term PBM (2 h) increases superoxide ion content, this was not observed for long exposure (24 h). Mitochondrial respirations were slightly decreased after PBM. In the islet transplantation model, both pre-illuminated and non-illuminated islets improved metabolic control in diabetic rats with a safety profile regarding the post-transplantation period. In summary, for the first time, long-term PBM exhibited safety in terms of cell viability, insulin secretion, energetic profiles in vitro, and post-transplantation period in vivo. Further investigation is warranted to explore PBM's protective effects under conditions of stress, aiding in the development of innovative approaches for cellular therapy.
Asunto(s)
Supervivencia Celular , Diabetes Mellitus Experimental , Células Secretoras de Insulina , Trasplante de Islotes Pancreáticos , Islotes Pancreáticos , Animales , Ratas , Células Secretoras de Insulina/metabolismo , Células Secretoras de Insulina/efectos de la radiación , Células Secretoras de Insulina/citología , Supervivencia Celular/efectos de la radiación , Trasplante de Islotes Pancreáticos/métodos , Islotes Pancreáticos/metabolismo , Islotes Pancreáticos/efectos de la radiación , Diabetes Mellitus Experimental/terapia , Diabetes Mellitus Experimental/metabolismo , Línea Celular , Mitocondrias/metabolismo , Mitocondrias/efectos de la radiación , Insulina/metabolismo , Secreción de Insulina/efectos de la radiación , Terapia por Luz de Baja Intensidad/métodos , Masculino , Adenosina Trifosfato/metabolismo , RatonesRESUMEN
Bioenergetic pathways uniquely support sarcomere function which, in turn, helps to maintain functional skeletal muscle (SKM) mass. Emerging evidence supports alcohol (EtOH)-induced bioenergetic impairments in SKM and muscle precursor cells. We performed a scoping review to synthesize existing evidence regarding the effects of EtOH on SKM bioenergetics. Eligible articles from six databases were identified, and titles, abstracts, and full texts for potentially relevant articles were screened against inclusion criteria. Through the search, we identified 555 unique articles, and 21 met inclusion criteria. Three studies investigated EtOH effects on the adenosine triphosphate (ATP)-phosphocreatine (PCr) system, twelve investigated EtOH effects on glycolytic metabolism, and seventeen investigated EtOH effects on mitochondrial metabolism. Despite increased ATP-PCr system reliance, EtOH led to an overall decrease in bioenergetic function through decreased expression and activity of glycolytic and mitochondrial pathway components. However, effects varied depending on the EtOH dose and duration, model system, and sample type. The results detail the EtOH-induced shifts in energy metabolism, which may adversely affect sarcomere function and contribute to myopathy. These findings should be used to develop targeted interventions that improve SKM bioenergetic function, and thus sarcomere function, in people with Alcohol Use Disorder (AUD). Key areas in need of further investigation are also identified.
Asunto(s)
Adenosina Trifosfato , Metabolismo Energético , Etanol , Músculo Esquelético , Metabolismo Energético/efectos de los fármacos , Humanos , Músculo Esquelético/metabolismo , Músculo Esquelético/efectos de los fármacos , Etanol/efectos adversos , Etanol/farmacología , Animales , Adenosina Trifosfato/metabolismo , Glucólisis/efectos de los fármacos , Mitocondrias/metabolismo , Mitocondrias/efectos de los fármacos , Fosfocreatina/metabolismoRESUMEN
In vitro production of porcine embryos is a complicated process that includes in vitro maturation (IVM), in vitro fertilization (IVF) and in vitro culture (IVC). Insufficient cytoplasmic maturation, slow zona reaction and improper embryo culture conditions will compromise the efficiency of porcine embryo production in vitro. Previous studies have shown that insulin-transferrin-selenium (ITS) in IVM or IVC medium could improve porcine oocyte maturation, decrease polyspermy fertilization and promote subsequent embryonic development in vitro. However, the effect of ITS both in IVM and IVC media on porcine embryo production in vitro hasn't been elucidated. In this study, we found that 1.0% ITS supplementation in IVM/IVC media promoted the expansion of cumulus cells, raised mitochondrial membrane potential, increased ATP content and reduced ROS level in matured oocytes, improved blastocyst rate and the cell number of blastocyst, simultaneously. In conclusion, the IVM/IVC media supplemented with 1.0% ITS can improve the efficiency of porcine embryo production in vitro.
Asunto(s)
Blastocisto , Medios de Cultivo , Técnicas de Cultivo de Embriones , Fertilización In Vitro , Técnicas de Maduración In Vitro de los Oocitos , Insulina , Potencial de la Membrana Mitocondrial , Selenio , Transferrina , Animales , Técnicas de Maduración In Vitro de los Oocitos/métodos , Selenio/farmacología , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Femenino , Porcinos/embriología , Técnicas de Cultivo de Embriones/métodos , Técnicas de Cultivo de Embriones/veterinaria , Fertilización In Vitro/métodos , Fertilización In Vitro/veterinaria , Blastocisto/efectos de los fármacos , Blastocisto/citología , Blastocisto/fisiología , Transferrina/farmacología , Medios de Cultivo/farmacología , Medios de Cultivo/química , Especies Reactivas de Oxígeno/metabolismo , Oocitos/efectos de los fármacos , Oocitos/fisiología , Oocitos/citología , Desarrollo Embrionario/efectos de los fármacos , Células del Cúmulo/efectos de los fármacos , Células del Cúmulo/citología , Adenosina Trifosfato/metabolismoRESUMEN
Pyroptosis plays an important role in maintenance of intestinal homeostasis, the abnormal activation of NOD-like receptor thermal protein domain-associated protein 3 (NLRP3) inflammasome can promote the event and development of ulcerative colitis (UC). Its protective effects such as inhibiting pyroptosis in various inflammation-related diseases have been demonstrated, but whether resveratrol (RES) can also alleviate the progression of the disease by inhibiting pyroptosis in UC and the mechanism have rarely been studied. In this study, lipopolysaccharide (LPS) combined with adenosine triphosphate (ATP) was used to induce HT29 human colon cancer cells to construct an intestinal epithelial cell pyroptosis and inflammation model in vitro to investigate the anti-inflammatory effect of RES, reveal the regulatory mechanism of RES on pyroptosis, and provide a new theoretical basis for the treatment of UC. In vitro experiences, HT29 cells were dividing into control group, LPS/ATP group, RES low-dose group, RES high-dose group, NF-κB inhibitor pyrrolidine dithiocarbamate group (PDTC group), and LPS/ATP+PDTC group. The mRNA expressions of pyroptosis-related indicators such as NLRP3, apoptosis-associated speck-like protein containing CARD (ASC), Caspase-1(CASP1), IL-18, IL-1ß, and inflammatory factors such as TNF-α and IL-6 were detected by qRT-PCR. The protein expressions of pyroptosis-related indicators NLRP3, ASC, CASP1, IL-18, IL-1ß, NF-κB-p65 in the nucleus, and IκBα and p-IκBα in the cytoplasm were detected by Western blot. Immunofluorescence saw the distribution and expression of NLRP3, ASC and NF-κB-p65 protein in each group. The morphology and degree of pyroptosis in each group were observed by transmission electron microscope. The results showed that compared with the control group, the pyroptosis-related proteins including NLRP3, ASC, CASP1, IL-18, IL-1ß, and inflammatory factors including TNF-α and IL-6 in the LPS/ATP group were increased, and LPS/ATP activated the activity of NF-κB signaling pathway. Compared with the LPS/ATP group, RES downregulated the expression of pyroptosis-related proteins and inflammatory factors in HT29 cells, and inhibited the activation of the NF-κB signaling pathway in HT29 cells pyroptosis. RES down-regulates the pyroptosis of HT29 cells induced by LPS/ATP and the expression of pyroptosis-related indicators NLRP3, ASC, CASP1, IL-18, IL-1ß and inflammatory factors TNF-α and IL-6 in the inflammatory response and inhibits the occurrence of pyroptosis. The mechanism is related to the inhibition of NF-κB pathway activity.
Asunto(s)
Adenosina Trifosfato , Inflamación , Lipopolisacáridos , Proteína con Dominio Pirina 3 de la Familia NLR , Piroptosis , Resveratrol , Humanos , Células HT29 , Resveratrol/farmacología , Lipopolisacáridos/inmunología , Piroptosis/efectos de los fármacos , Adenosina Trifosfato/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Inflamación/metabolismo , Inflamación/tratamiento farmacológico , Inflamación/inmunología , Inflamasomas/metabolismo , Inflamasomas/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , FN-kappa B/metabolismo , Colitis Ulcerosa/tratamiento farmacológico , Colitis Ulcerosa/inmunología , Colitis Ulcerosa/metabolismo , Colitis Ulcerosa/patología , Colitis Ulcerosa/inducido químicamenteRESUMEN
Proteins occurring in significantly high concentrations in cellular environments (over 100 mg/ml) and functioning in crowded cytoplasm, often face the prodigious challenges of aggregation which are the pathological hallmark of aging and are critically responsible for a wide spectrum of rising human diseases. Here, we combine a joint-venture of complementary wet-lab experiment and molecular simulation to discern the potential ability of adenosine triphosphate (ATP) as solubilizer of protein aggregates. We show that ATP prevents both condensation of aggregation-prone intrinsically disordered protein Aß40 and promotes dissolution of preformed aggregates. Computer simulation links ATP's solubilizing role to its ability to modulate protein's structural plasticity by unwinding protein conformation. We show that ATP is positioned as a superior biological solubilizer of protein aggregates over traditional chemical hydrotropes, potentially holding promises in therapeutic interventions in protein-aggregation-related diseases. Going beyond its conventional activity as energy currency, the amphiphilic nature of ATP enables its protein-specific interaction that would enhance ATP's efficiency in cellular processes.
Asunto(s)
Adenosina Trifosfato , Agregado de Proteínas , Solubilidad , Adenosina Trifosfato/metabolismo , Humanos , Conformación Proteica , Simulación por ComputadorRESUMEN
OBJECTIVE: The objective of this study was to investigate the protective mechanism of Ling-Gui-Zhu-Gan decoction (LGZGD) against LPS-ATP-induced pyroptosis in H9c2 cells. METHODS: LPS and ATP were used to induce pyroptosis in the H9c2 cell, and the cells were divided into the control, model and LGZGD groups. LDH level was detected using a colorimetric assay. ELISA was used to detect the expressions of IL-1ß. Flow cytometry was utilized to observe apoptosis, while Hoechst/PI staining was used to detect pyroptosis. Immunofluorescence was employed to observe the expression levels of NLRP3 in cardiomyocytes, and RT-PCR was used to detect NLRP3, Caspase-1, GSDMD, and ASC mRNA expression. The cells were separated into seven groups: control, model, LGZGD, MCC950, LGZGD+MCC950, Nigericin and LGZGD+Nigericin. The mRNA and protein expressions were determined by RT-PCR and Western blot. RESULTS: LPS (10⯵g/mL) for 12â¯h and ATP (8â¯mM) for 2â¯h were used as modeling condition. LGZGD demonstrated a significant reduction in LDH, and IL-1ß levels (P<0.05, P<0.01). LGZGD dramatically reduced apoptosis rate, inhibited pyroptosis, decreased the fluorescence expressions of NLRP3, and reduced the mRNA expressions of NLRP3, ASC, Caspase-1, and GSDMD (P<0.01). Further mechanism studies showed that NLRP3, ASC, Caspase-1, and GSDMD decreased significantly when combined with NLRP3 inhibitor MCC950. Furthermore, LGZGD was able to effectively reverse the upregulation of protein and gene expression of Nigericin group (P<0.01). CONCLUSION: LGZGD inhibits LPS-ATP-induced pyroptosis in H9c2 cell via the NLRP3/Caspase-1 signaling pathway.
Asunto(s)
Caspasa 1 , Medicamentos Herbarios Chinos , Lipopolisacáridos , Miocitos Cardíacos , Proteína con Dominio Pirina 3 de la Familia NLR , Piroptosis , Transducción de Señal , Piroptosis/efectos de los fármacos , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Caspasa 1/metabolismo , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Transducción de Señal/efectos de los fármacos , Animales , Medicamentos Herbarios Chinos/farmacología , Ratas , Línea Celular , Lipopolisacáridos/farmacología , Interleucina-1beta/metabolismo , Adenosina Trifosfato/metabolismoRESUMEN
Pyruvate serves as a key metabolite in energy production and as an anti-oxidant. In our previous study, exogenous pyruvate starvation under high-glucose conditions induced IMS32 Schwann cell death because of the reduced glycolysis-tricarboxylic acid (TCA) cycle flux and adenosine triphosphate (ATP) production. Thus, this study focused on poly-(ADP-ribose) polymerase (PARP) to investigate the detailed molecular mechanism of cell death. Rucaparib, a PARP inhibitor, protected Schwann cells against cell death and decreased glycolysis but not against an impaired TCA cycle under high-glucose conditions in the absence of pyruvate. Under such conditions, reduced pyruvate dehydrogenase (PDH) activity and glycolytic and mitochondrial ATP production were observed but not oxidative phosphorylation or the electric transfer chain. In addition, rucaparib supplementation restored glycolytic ATP production but not PDH activity and mitochondrial ATP production. No differences in the increased activity of caspase 3/7 and the localization of apoptosis-inducing factor were found among the experimental conditions. These results indicate that Schwann cells undergo necrosis rather than apoptosis or parthanatos under the aforementioned conditions. Exogenous pyruvate plays a pivotal role in maintaining the flux in PARP-dependent glycolysis and the PARP-independent TCA cycle in Schwann cells under high-glucose conditions.
Asunto(s)
Adenosina Trifosfato , Ciclo del Ácido Cítrico , Glucosa , Glucólisis , Poli(ADP-Ribosa) Polimerasas , Ácido Pirúvico , Células de Schwann , Glucólisis/efectos de los fármacos , Ciclo del Ácido Cítrico/efectos de los fármacos , Adenosina Trifosfato/metabolismo , Glucosa/metabolismo , Ácido Pirúvico/metabolismo , Animales , Células de Schwann/metabolismo , Células de Schwann/efectos de los fármacos , Poli(ADP-Ribosa) Polimerasas/metabolismo , Mitocondrias/metabolismo , Mitocondrias/efectos de los fármacos , Ratas , Inhibidores de Poli(ADP-Ribosa) Polimerasas/farmacología , Indoles/farmacología , Línea Celular , Apoptosis/efectos de los fármacosRESUMEN
Ginseng has anti-hyperglycemic effects. Gintonin, a glycolipoprotein derived from ginseng, also stimulates insulin release from pancreatic beta cells. However, the role of gintonin in glucose metabolism within skeletal muscle is unknown. Here, we showed the effect of gintonin on glucose uptake, glycogen content, glucose transporter (GLUT) 4 expression, and adenosine triphosphate (ATP) content in C2C12 myotubes. Gintonin (3-30 µg/mL) dose-dependently stimulated glucose uptake in myotubes. The expression of GLUT4 on the cell membrane was increased by gintonin treatment. Treatment with 1-3 µg/mL of gintonin increased glycogen content in myotubes, but the content was decreased at 30 µg/mL of gintonin. The ATP content in myotubes increased following treatment with 10-100 µg/mL gintonin. Gintonin transiently elevated intracellular calcium concentrations and increased the phosphorylation of extracellular signal-regulated kinase (ERK). Gintonin-induced transient calcium increases were inhibited by treatment with the lysophosphatidic acid receptor inhibitor Ki16425, the phospholipase C inhibitor U73122, and the inositol 1,4,5-trisphosphate receptor antagonist 2-aminoethoxydiphenyl borate. Gintonin-stimulated glucose uptake was decreased by treatment with U73122, the intracellular calcium chelator 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetra(acetoxymethyl) ester, and the ERK inhibitor PD98059. These results show that gintonin plays a role in glucose metabolism by increasing glucose uptake through transient calcium increases and ERK signaling pathways. Thus, gintonin may be beneficial for glucose metabolism control.
Asunto(s)
Calcio , Transportador de Glucosa de Tipo 4 , Glucosa , Fibras Musculares Esqueléticas , Glucosa/metabolismo , Animales , Ratones , Calcio/metabolismo , Transportador de Glucosa de Tipo 4/metabolismo , Fibras Musculares Esqueléticas/metabolismo , Fibras Musculares Esqueléticas/efectos de los fármacos , Adenosina Trifosfato/metabolismo , Línea Celular , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Glucógeno/metabolismo , Extractos VegetalesRESUMEN
OBJECTIVE: To assess the potential of bypassing mitochondrial complex I with idebenone to overcome drug resistance in a Rotenone corneal kindling (RCK) mouse model of mitochondrial refractory epilepsy. MATERIAL AND METHOD: Resistance was developed by administering rotenone 2.5â¯mg/kg intraperitoneally once and corneal kindling twice daily. The kindling development took 15 days, and pre-treatment resistance validation was carried out with five different antiseizure drugs: pregabalin, levetiracetam, valproate, lamotrigine, and phenytoin. The treatment drug, Idebenone (IDB) was given at doses of 10, 20, and 40â¯mg/kg intraperitoneally for 10 days. The post-treatment resistance validation was evaluated with same standard drugs in same order along with other parameters assessment, such as NAD(P)H: quinone oxidoreductase 1 (NQO1), ATP, GSH, and TBARS. RESULTS: The pre-treatment resistance validation shows an inability of standard drugs to attenuate seizure scores by rotenone kindling, justifying the development of drug resistance. IDB successfully abolished the resistance developed in RCK model. IDB elevated the levels of ATP and NQO1 and showed antioxidant activity by elevating GSH and attenuating TBARS. CONCLUSION & FUTURE DIRECTION: IDB have successfully elevated the level of ATP, NQO1 in RCK model, hence proving the complex I bypass hypothesis. Thus, IDB can be the drug of choice for mitochondrial epilepsies involving drug refractoriness as adjuvant with anticonvulsant drugs.