RESUMEN
Adagrasib (MRTX849), an approved and promising KRAS G12C inhibitor, has shown the promising results for treating patients with advanced non-small cell lung cancer (NSCLC) or colorectal cancer (CRC) harboring KRAS-activating mutations. However, emergence of the acquired resistance limits its long-term efficacy and clinical application. Further understanding of the mechanism of the acquired resistance is crucial for developing more new effective therapeutic strategies. Herein, we firstly found a new connection between the acquired resistance to MRTX849 and nuclear factor erythroid 2-related factor 2 (Nrf2). The expression levels of Nrf2 and GLS1 proteins were substantially elevated in different CRC cell lines with the acquired resistance to MRTX849 in comparison with their corresponding parental cell lines. Next, we discovered that RA-V, one of natural cyclopeptides isolated from the roots of Rubia yunnanensis, could restore the response of resistant CRC cells to MRTX849. The results of molecular mechanisms showed that RA-V suppressed Nrf2 protein through the ubiquitin-proteasome-dependent degradation, leading to the induction of oxidative and ER stress, and DNA damage in CRC cell lines. Consequently, RA-V reverses the resistance to MRTX849 by inhibiting the Nrf2/GLS1 axis, which shows the potential for further developing into one of novel adjuvant therapies of MRTX849.
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Neoplasias Colorrectales , Factor 2 Relacionado con NF-E2 , Péptidos Cíclicos , Factor 2 Relacionado con NF-E2/metabolismo , Factor 2 Relacionado con NF-E2/genética , Humanos , Neoplasias Colorrectales/tratamiento farmacológico , Neoplasias Colorrectales/metabolismo , Neoplasias Colorrectales/patología , Neoplasias Colorrectales/genética , Línea Celular Tumoral , Péptidos Cíclicos/farmacología , Péptidos Cíclicos/uso terapéutico , Resistencia a Antineoplásicos/efectos de los fármacos , Animales , Antineoplásicos Fitogénicos/farmacología , Ratones DesnudosRESUMEN
The object of this study was to trace TwHf-derived toxins in raw honey and clarify their acute toxic effect related to the addition of honey or sugars. TwHf flowers, raw honey from TwHf planting base and from beekeepers in high-risk area were detected using LC-MS/MS. The results revealed five target toxins were detected in TwHf flowers; only celastrol was detected in one raw honey sample, as a food safety risk factor, celastrol had been traced back to TwHf flowers from raw honey. In a series of acute toxic tests on zebrafish, toxification effects were observed when honey, mimic honey or sugar was mixed with toxins. The degree of toxicity varied among various sugar-based solutions. At the same mass concentration, they follow this order: raw honey/mimic honey > glucose > fructose. The main toxic target organs of triptolide and celastrol with honey were the heart and liver.
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Diterpenos , Miel , Triterpenos Pentacíclicos , Fenantrenos , Tripterygium , Animales , Miel/análisis , Cromatografía Liquida , Pez Cebra , Espectrometría de Masas en Tándem , Azúcares , Compuestos EpoxiRESUMEN
This study demonstrates that the purified ß-glucan (LNT) with a triple helix and relatively narrow molecular weight distribution, extracted and purified from artificially cultured Lentinus edodes, showed a significant cervical cancer inhibition with little cytotoxicity against normal cells in vitro and in vivo. From the in vitro data, the potential mechanism of anti-cervical cancer was preliminarily revealed as follows: LNT was firstly recognized by the human cervical cancer cell line of Hela and induced cell proliferation inhibition through p21 and apoptosis via a mitochondrion-dependent pathway by targeting the tumor suppressor of p53, indicated by an increase in reactive oxygen species (ROS) generation and a loss of mitochondrial membrane potential (Δψm), in a significant dosage-dependent manner. Meanwhile, LNT repressed tumor growth with an inhibition ratio of 61.2 % and induced tumor cell apoptosis through endogenous MDM2/p53/Bax/mitochondrion signal pathway by up-regulating the expression of p53, Bax, Cyt. c, caspase 9, and caspase 3, as well as down-regulating Bcl-2, MDM2, and PARP1 levels in Hela cells-transplanted BALB/c nude mice. This study provides a scientific basis for the clinical treatment of cervical cancer with LNT as a potential drug candidate characterized by the triple helix and specified molecular weight with a relatively narrow distribution.
RESUMEN
Lipophagy is the autophagic degradation of lipid droplets. Dysregulated lipophagy has been implicated in the development of non-alcoholic fatty liver disease (NAFLD). Ajugol is an active alkaloid isolated from the root of Rehmannia glutinosa which is commonly used to treat various inflammatory and metabolic diseases. This study aimed to investigate the effect of ajugol on alleviating hepatic steatosis and sought to determine whether its potential mechanism via the key lysosome-mediated process of lipophagy. Our findings showed that ajugol significantly improved high-fat diet-induced hepatic steatosis in mice and inhibited palmitate-induced lipid accumulation in hepatocytes. Further analysis found that hepatic steatosis promoted the expression of LC3-II, an autophagosome marker, but led to autophagic flux blockade due to a lack of lysosomes. Ajugol also enhanced lysosomal biogenesis and promoted the fusion of autophagosome and lysosome to improve impaired autophagic flux and hepatosteatosis. Mechanistically, ajugol inactivated mammalian target of rapamycin and induced nuclear translocation of the transcription factor EB (TFEB), an essential regulator of lysosomal biogenesis. siRNA-mediated knockdown of TFEB significantly abrogated ajugol-induced lysosomal biogenesis as well as autophagosome-lysosome fusion and lipophagy. We conclude that lysosomal deficit is a critical mediator of hepatic steatosis, and ajugol may alleviate NAFLD via promoting the TFEB-mediated autophagy-lysosomal pathway and lipophagy.
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Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/metabolismo , Glicósidos Iridoides/uso terapéutico , Enfermedad del Hígado Graso no Alcohólico/tratamiento farmacológico , Piranos/uso terapéutico , Animales , Autofagia/efectos de los fármacos , Células Cultivadas , Dieta Alta en Grasa , Hepatocitos/efectos de los fármacos , Hepatocitos/metabolismo , Glicósidos Iridoides/farmacología , Metabolismo de los Lípidos/efectos de los fármacos , Lisosomas/efectos de los fármacos , Masculino , Ratones Endogámicos C57BL , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Obesidad/tratamiento farmacológico , Obesidad/metabolismo , Piranos/farmacologíaRESUMEN
The Rotimer, a rotifer-specific biopolymer, is an exogenic bioactive exudate secreted by different monogonant species (e.g. Euchlanis dilatata or Lecane bulla). The production of this viscoelastic biomolecule is induced by different micro-particles, thereby forming a special Rotimer-Inductor Conglomerate (RIC) in a web format. In this case, the water insoluble Carmine crystals, filtered to size (max. diameter was 50 µm), functioned as an inductor. The RIC production is an adequate empirical indicator to follow up this filamentous biopolymer secretion experientially; moreover, this procedure is very sensitive to the environmental factors (temperature, pH, metals and possible natural pollutant agents). The above mentioned species show completely different reactions to these factors, except to the presence of calcium and to the modulating effects of different drugs. One of the novelties of this work is that the Rotimer secretion and consequently, the RIC-formation is a mutually obligatory and evolutionary calcium-dependent process in the concerned monogonants. This in vivo procedure needs calcium, both for the physiology of animals and for fiber formation, particularly in the latter case. The conglomerate covered area (%) and the detection of the longest filament (mm) of the given RIC were the generally and simultaneously applied methods in the current modulating experiments. Exploring the regulatory (e.g. calcium-dependency) and stimulating (e.g. Lucidril effect) possibilities of biopolymer secretion are the basis for optimizing the RIC-production capacities of these micro-metazoans.
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Biopolímeros/biosíntesis , Calcio/farmacología , Ambiente , Contaminantes Ambientales/farmacología , Exudados y Transudados , Rotíferos/metabolismo , Animales , Concentración de Iones de Hidrógeno , Rotíferos/efectos de los fármacos , TemperaturaRESUMEN
The chemical ecology of rotifers has been little studied. A yet unknown property is presented within some monogonant rotifers, namely the ability to produce an exogenic filamentous biopolymer, named 'Rotimer'. This rotifer-specific viscoelastic fiber was observed in six different freshwater monogonants (Euchlanis dilatata, Lecane bulla, Lepadella patella, Itura aurita, Colurella adriatica and Trichocerca iernis) in exception of four species. Induction of Rotimer secretion can only be achieved by mechanically irritating rotifer ciliate with administering different types (yeast cell skeleton, denatured BSA, epoxy, Carmine or urea crystals and micro-cellulose) and sizes (approx. from 2.5 to 50 µm diameter) of inert particles, as inductors or visualization by adhering particles. The thickness of this Rotimer is 33 ± 3 nm, detected by scanning electron microscope. This material has two structural formations (fiber or gluelike) in nano dimension. The existence of the novel adherent natural product becomes visible by forming a 'Rotimer-Inductor Conglomerate' (RIC) web structure within a few minutes. The RIC-producing capacity of animals, depends on viability, is significantly modified according to physiological- (depletion), drug- (toxin or stimulator) and environmental (temperature, salt content and pH) effects. The E. dilatata-produced RIC is affected by protein disruptors but is resistant to several chemical influences and its Rotimer component has an overwhelming cell (algae, yeast and human neuroblastoma) motility inhibitory effect, associated with low toxicity. This biopolymer-secretion-capacity is protective of rotifers against human-type beta-amyloid aggregates.
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Biopolímeros/metabolismo , Rotíferos/metabolismo , Péptidos beta-Amiloides/farmacología , Animales , Biopolímeros/química , Biopolímeros/farmacología , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Agua Dulce/microbiología , Humanos , Rotíferos/clasificación , Rotíferos/efectos de los fármacos , TemperaturaRESUMEN
BACKGROUND AND PURPOSE: Inflammation has been considered a precipitating event that contributes to neurocognitive dysfunction in minimal hepatic encephalopathy (MHE). Inhibition TLR-4 related inflammation can effectively improve neurocognitive dysfunction of MHE. Our previous study showed that Babao Dan (BBD) effectively inhibited inflammation and ameliorated neurocognitive function in rats with acute hepatic encephalopathy (HE) and chronic HE. The mechanism may lie in the regulation of TLR4 signaling pathway. Therefore, this study aimed to evaluate the role of BBD in the treatment of MHE patients with cirrhosis and to elucidate the underlying mechanism by which BBD regulated TLR4 pathway to alleviate inflammation. METHODS: A randomized controlled trial (n = 62) was conducted to evaluate the clinical efficacy between BBD plus lactulose (n = 31) and lactulose alone (n = 31) in MHE patients by testing neurocognitive function (NCT-A and DST), blood ammonia, liver function (ALT, AST and TBIL) and blood inflammation (IL-1ß, IL-6 and TNF-α). Afterward, we detected NO, inflammatory cytokines (IL-1ß, IL-6 and TNF-α) and the phosphorylation of P65, JNK, ERK as well as P38 in LPS-activated rat primary bone marrow-derived macrophages (BMDMs), peritoneal macrophages (PMs), and mouse primary BMDMs/PMs/microglia/astrocytes, to investigate the underlying mechanism of BBD inhibiting inflammation through TLR4 pathway. Also, the survival rate of mice, liver function (ALT, AST), blood inflammation (IL-1ß, IL-6 and TNF-α), inflammatory cytokines (IL-1ß, IL-6 and TNF-α) and histopathological changes in the liver, brain and lung were measured to assess the anti-inflammatory effect of BBD on neurocognitive function in endotoxin shock/endotoxemia mice. RESULTS: BBD combined with lactulose significantly ameliorated neurocognitive function by decreasing NCT-A (pï¼0.001) and increasing DST (pï¼0.001); inhibited systemic inflammation by decreasing IL-1ß (pï¼0.001), IL-6(pï¼0.001) and TNF-α (pï¼0.001); reduced ammonia level (p = 0.005), and improved liver function by decreasing ALT(p = 0.043), AST(p = 0.003) and TBIL (p = 0.026) in MHE patients. Furthermore, BBD inhibited gene and protein expression of IL-1ß, IL-6 and TNF-α as well as NO in rat primary BMDMs/PMs, and mouse primary BMDMs/PMs/microglia/astrocytes in a dose-dependent manner. BBD inhibited the activation of mouse primary BMDMs/PMs/microglia/astrocytes by regulating TLR4 pathway involving the phosphorylation of P65, JNK, ERK and P38. Also, BBD reduced the mortality of mice with endotoxin shock/endotoxemia; serum levels of ALT, AST, IL-1ß, IL-6 and TNF-α; gene expression of IL-1ß, IL-6 and TNF-α in the liver, brain and lung, and tissue damage in the liver and lung. CONCLUSION: Our study provided for the first time clinical and experimental evidence supporting the use of BBD in MHE, and revealed that BBD could play a crucial role in targeting and regulating TLR4 inflammatory pathway to improve neurocognitive function in MHE patients.
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Antiinflamatorios , Encéfalo , Cognición , Citocinas , Medicamentos Herbarios Chinos , Encefalopatía Hepática , Mediadores de Inflamación , Anciano , Animales , Femenino , Humanos , Masculino , Persona de Mediana Edad , Embarazo , Antiinflamatorios/efectos adversos , Antiinflamatorios/uso terapéutico , Astrocitos/efectos de los fármacos , Astrocitos/metabolismo , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Encéfalo/fisiopatología , Células Cultivadas , China , Cognición/efectos de los fármacos , Citocinas/metabolismo , Modelos Animales de Enfermedad , Medicamentos Herbarios Chinos/efectos adversos , Medicamentos Herbarios Chinos/uso terapéutico , Endotoxemia/tratamiento farmacológico , Endotoxemia/metabolismo , Encefalopatía Hepática/tratamiento farmacológico , Encefalopatía Hepática/metabolismo , Encefalopatía Hepática/fisiopatología , Encefalopatía Hepática/psicología , Mediadores de Inflamación/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Ratones Endogámicos C57BL , Microglía/efectos de los fármacos , Microglía/metabolismo , Factores de Tiempo , Receptor Toll-Like 4/metabolismo , Resultado del Tratamiento , RatonesRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Oroxyloside is a natural flavonoid isolated from Scutellaria baicalensis Georgi (Lamiaceae) which is a Chinese herb widely used for liver diseases. However, its mechanisms on protecting against drug induced liver injury has not been investigated yet. AIM OF THE STUDY: To investigate the protecting effects and the primary mechanisms of oroxyloside on acetaminophen (APAP)-induced liver injury. MATERIALS AND METHODS: After a 12 h fasting period with free access to water, C57BL/6 mice were injected with APAP (300 mg/kg) intragastrically (i.g.) and 1 h later with oroxyloside (100 mg/kg, i.g.). When mice sacrificed, blood samples were collected from fundus venous plexus and liver tissues were collected. In addition, cells were incubated with 10 mM APAP alone and 10 mM APAP combined with 100 µM oroxyloside for 24 h. ELISA, TUNEL assay, qRT-PCR et al. were used to assess the effect of oroxyloside on ameliorating APAP-induced hepatotoxicity in vitro and in vivo. Western bolt and immunohistochemistry were used in the signaling pathway analysis. RESULTS: Oroxyloside administration significantly decreased the accumulations of CYP2E1, CYP1A2, IL-6, IL-1ß, ALT and AST induced by APAP in vivo. In addition, oroxyloside inhibited the APAP-induced JNK related apoptosis by enhancing the antioxidant defenses, reversing ER-stress and keeping the mito-balance of liver cells in vivo and in vitro. Furthermore, oroxyloside protected the liver cells from necroptosis by affecting JNK pathway. CONCLUSION: Oroxyloside acted as a protective agent against APAP-induced liver injury through inhibiting JNK-related apoptosis and necroptosis.
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Acetaminofén/toxicidad , Apoptosis/efectos de los fármacos , Enfermedad Hepática Inducida por Sustancias y Drogas/prevención & control , Flavonas/farmacología , Glucurónidos/farmacología , Animales , Antioxidantes/metabolismo , Enfermedad Hepática Inducida por Sustancias y Drogas/etiología , Femenino , Flavonas/aislamiento & purificación , Glucurónidos/aislamiento & purificación , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Ratones , Ratones Endogámicos C57BL , Necroptosis/efectos de los fármacos , Scutellaria baicalensis/química , Transducción de Señal/efectos de los fármacosRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: BabaoDan (BBD) is a famous traditional Chinese formula frequently used in TCM clinics to eliminate jaundice and treat infectious viral hepatitis. This paper assesses BBD's preventive and therapeutic effects on hepatic encephalopathy after liver cirrhosis (CHE) and acute liver failure (AHE) in rats and explains its possible mechanism of action. METHODS: CHE rat model was established by injection of carbon tetrachloride (CCl4) twice a week for a total of 9 weeks and then by injection of thioacetamide (TAA) to induce hepatic encephalopathy. AHE rat model was established by injection of TAA once a day for a total of 3 days. In CHE rat model, BBD was gavaged once a day at the end of the 6th week until the experiment ended. In AHE rat model,BBD was gavaged once a day 3 days before TAA injection until the experiment ended. The preventive and therapeutic effects of BBD on brain dysfunction, as well as liver injury, pathology and fibrosis were evaluated in vivo. The role of BBD in the regulation of inflammatory factors and myeloid differentiation factor 88/Toll-like receptor 4/nuclear factor kappa-B (TLR4/MyD88/NK-κ B) pathway was detected in both liver and brain in vivo. The rat bone marrow derived macrophages (BMDMs) were activated by Lipopolysaccharide (LPS), and the role of BBD in the regulation of inflammatory factors and NK-κ B pathway were detected in vitro. RESULTS: In CHE rat model: BBD significantly improved the total distance as well as the activity rate of rats. BBD also improved the learning and memory abilities of rats compared with the control group. In addition, BBD effectively decreased ammonia levels and significantly decreased the levels of alanine aminotransferase (ALT), aspartate transaminase (AST), total bilirubin (TBil) and total bile acid (TBA), as well as improved the levels of total protein (TP) and albumin (Alb). In the liver, BBD not only inhibited the gene expressions of tumor necrosis factor alpha (TNF-α), interleukini-6 (IL-6), TLR4, MyD88, and NF-κ B but also inhibited the protein expressions of TLR4, MyD88, NK-κ B and TNF-α. In the brain, BBD inhibited the gene expressions of iNOS, IL-6, TNF-α, TLR-4, MyD88, and NF-κ B, as well as inhibited the protein expressions of TLR4, MyD88, P65 TNF-α and ionized calcium binding adapter molecule 1 (Iba-1). BBD also decreased NO and TNF-α in the blood. IN AHE RAT MODEL: BBD improved neurological scores, blood ammonia levels and the brain inflammatory gene expressions of iNOS, TNF-α and IL-1ß. BBD also improved liver function biomarkers such as ALT, TBil, TBA, TP, ALB and inflammatory and apoptotic gene expressions of TNF-α, IL-1ß, IL-6, Bax, Bcl-2, caspase-9, caspase-3 and NF-κ B. In LPS-activated rat BMDMs, BBD decreased NO and TNF-α production in BMDM culture supernatant. In addition, BBD inhibited the gene expressions of TNF-α, IL-1 ß and IL-6 as well as the phosphorylation of P65. CONCLUSION: BBD can prevent and cure hepatic encephalopathy (HE) derived from both chronic and acute liver diseases. BBD can reduce hyperammonemia as well as the systematic and neurological inflammation. Inflammation is likely an important target of BBD to treat HE. The anti-inflammatory role of BBD may lie in its regulation of the TLR4/MyD88/NF-κ B pathways.
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Amoníaco/metabolismo , Antiinflamatorios/farmacología , Encefalopatía Hepática/tratamiento farmacológico , Inflamación/tratamiento farmacológico , Hígado/efectos de los fármacos , Animales , Modelos Animales de Enfermedad , Encefalopatía Hepática/metabolismo , Inflamación/metabolismo , Hígado/metabolismo , Fallo Hepático Agudo/tratamiento farmacológico , Fallo Hepático Agudo/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Masculino , Factor 88 de Diferenciación Mieloide/metabolismo , FN-kappa B/metabolismo , Ratas , Ratas Wistar , Transducción de Señal/efectos de los fármacos , Receptor Toll-Like 4/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Soybean Bowman-Birk trypsin inhibitor (BBTI), an antinutritional factor of soy products, could strongly inhibit the protein digestion. The inactivation effect and mechanism of BBTI induced by tea polyphenols (TPs) and its major components (EGCG and EGC), were investigated in this study using fluorescence, FTIR, CD spectroscopy, isothermal titration calorimetry (ITC) and molecular docking. EGCG and EGC interacted with BBTI via static quenching process and hydrophobic interaction, with binding constant (Ka) of 2.19â¯×â¯103â¯M-1 and 0.25â¯×â¯103â¯M-1 at 298â¯K, respectively. TPs, EGCG and EGC induced a transition of BBTI conformation from disorder to order. ITC analysis and molecular docking revealed the interaction of EGCG-BBTI and EGC-BBTI were spontaneous, and hydrophobic interactions and hydrogen bonds were the predominant forces. Overall, this study clearly suggested that EGCG could be a promising inactivating agent for BBTI, which could also improve the safety and nutritional value of soy products.
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Catequina/análogos & derivados , Simulación del Acoplamiento Molecular , Inhibidor de la Tripsina de Soja de Bowman-Birk/química , Catequina/química , Catequina/farmacología , Fluorescencia , Enlace de Hidrógeno , Interacciones Hidrofóbicas e Hidrofílicas , Conformación Proteica , Termodinámica , Inhibidor de la Tripsina de Soja de Bowman-Birk/efectos de los fármacos , Inhibidor de la Tripsina de Soja de Bowman-Birk/metabolismoRESUMEN
In this study, we prepared the platelet-derived growth factor-containing porous microspheres modified with heparin (PDGF/Hep-PMSs) and investigated their anti-inflammatory and tendon healing effects on rotator cuff (RC) tendinitis rabbit model. PDGF/Hep-PMSs suppressed the mRNA levels of six pro-inflammatory cytokines (i.e., MMP-3, MMP-13, COX-2, ADAMTS-5, IL-6, and TNF-α) in inflamed tenocytes. Long-term local delivery of PDGF/Hep-PMSs into tendon tissues of RC tendinitis decreased the mRNA levels of six pro-inflammatory cytokines and increased the mRNA levels of anti-inflammatory cytokines including IL-4, IL-10, and IL-13. Anti-inflammatory effects of PDGF/Hep-PMSs might have contributed to enhance the collagen content, tenogenic markers, stiffness, and tensile strength of tendons, eventually leading to tendon restoration. Our findings suggest that the long-term local PDGF delivery of PDGF/Hep-PMSs have a great potential to enhance tendon healing of RC tendinitis by suppressing inflammation responses.
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Portadores de Fármacos/química , Heparina/química , Microesferas , Factor de Crecimiento Derivado de Plaquetas/química , Factor de Crecimiento Derivado de Plaquetas/farmacología , Lesiones del Manguito de los Rotadores/tratamiento farmacológico , Tendones/efectos de los fármacos , Animales , Proliferación Celular/efectos de los fármacos , Modelos Animales de Enfermedad , Dopamina/química , Liberación de Fármacos , Factor de Crecimiento Derivado de Plaquetas/uso terapéutico , Copolímero de Ácido Poliláctico-Ácido Poliglicólico/química , Porosidad , Conejos , Lesiones del Manguito de los Rotadores/patología , Lesiones del Manguito de los Rotadores/fisiopatología , Propiedades de Superficie , Tendones/patología , Tendones/fisiopatología , Factores de TiempoRESUMEN
Ten barley samples containing varied protein contents were subject to malting followed by mashing to investigate molecular effects of both barley starch and starch- protein interactions on malting and mashing performances, and the underlying mechanism. Starch granular changes were examined using differential scanning calorimetry and scanning electron microscopy. The molecular fine structures of amylose and amylopectin from unmalted and malted grain were obtained using size-exclusion chromatography. The results showed that both amylose and amylopectin polymers were hydrolyzed at the same time during malting. Protein and amylose content in both unmalted and malted barley significant negatively correlated with fermentable sugar content after mashing. While protein content is currently the main criterion for choosing malting varieties, this study shows that information about starch molecular structure is also useful for determining the release of fermentable sugars, an important functional property. This provides brewers with some new methods to choose malting barley.
RESUMEN
Medicinal plants and herbal extracts from traditional Chinese medicine are used increasingly commonly worldwide for their benefits to health and quality of life as dietary supplements or as ingredients in functional foods. Among them, Salvia miltiorrhiza Bunge (a natural strong remedy for the treatment of a variety of conditions) is traditionally used for centuries in Asian countries as antioxidant, anticancer, and anti-inflammatory agent. In this context, several evidences support the hypothesis that some tanshinones (in particular tanshinone IIA and cryptotanshinone) extracted from the roots (Danshen) of Salvia miltiorrhiza exert neuroprotective and analgesic activities. Oxaliplatin (OXA), a platinum-based drug used for the treatment of solid tumors, induces neuropathic pain which hampers the chemotherapy success. While several attempts were made to prevent oxaliplatin-induced painful neuropathy, a growing number of evidences look to natural sources as an effective remedy to counterbalance the OXA-mediated side effects. The aim of the present study was to investigate the pain-relieving profile of Danshen and its active constituents tanshinone IIA (TIIA) and cryptotanshinone (CRY) in animal models of neuropathic pain induced by OXA, anticancer drug characterized by a dose-limiting neurotoxicity. Contextually, the neuroprotective and anticancer activities of the selected compounds were tested in different cells lines. A single administration per os of CRY (30â¯mg mg/kg) significantly, in a dose dependent manner, attenuated chemotherapy-induced pain. A 7 days repeated administrations highlighted the effectiveness and potency of both CRY and TIIA (10â¯mg/kg). On the other hand, Danshen showed a painkiller profile against oxaliplatin-induced neuropathy. Contextually, Danshen and its active constituents showed remarkable and selective inhibitory activities on glioblastoma cells lines LN-229 (IC50: 50.0⯱â¯4.0, 48.2⯱â¯4.9 and 51.9⯱â¯2.3⯵M respectively for Danshen standardized extract, TIIA and CRY) next to healthy but high proliferative cell lines enterocytes (IC50:> 250⯵M for TIIA and CRY) and keratinocytes (IC50: >100 and 97⯱â¯2⯵M respectively for TIIA and CRY). Taken together the results reported here demonstrated the long-lasting pain-relieving effects of Danshen and its related bioactive constituents in animal models of neuropathic pain and their selective in vitro neuroprotective properties on certain central malignancy cells lines. Thus, suggest that S. miltiorrhiza roots could be considered as a new potential source of active diterpenoidic compounds useful for pharmaceutical or nutraceutical industries and beneficial as food complements.
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Abietanos/uso terapéutico , Antineoplásicos/toxicidad , Neoplasias Encefálicas/tratamiento farmacológico , Glioblastoma/tratamiento farmacológico , Neuralgia/tratamiento farmacológico , Salvia miltiorrhiza , Abietanos/aislamiento & purificación , Abietanos/farmacología , Animales , Neoplasias Encefálicas/patología , Línea Celular Tumoral , Relación Dosis-Respuesta a Droga , Medicamentos Herbarios Chinos/aislamiento & purificación , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/uso terapéutico , Glioblastoma/patología , Humanos , Masculino , Ratones , Neuralgia/inducido químicamente , Neuralgia/patología , Dimensión del Dolor/efectos de los fármacos , Dimensión del Dolor/métodosRESUMEN
Hyperactivated macrophages play a key role in the initiation and perpetuation of mucosal inflammation in Crohn's disease (CD). Increasing evidence suggests that the basic helix-loop-helix (bHLH) repressor Twist1 can suppress activation of nuclear factor-κB (NF-κB) and the subsequent production of TNF-α, which are both essential elements of macrophage activation. Thus, developing novel therapeutic strategies to enhance Twist1 expression and to inhibit macrophage activation may be beneficial for CD treatment. In the present study, a series of trifluoroethyl thiazolo[3,2-b][1,2,4]triazole derivatives were used to investigate their potential anti-inflammatory activities and the underlying mechanism. In a biological activity screen, compound 7# (Thiazolo[3,2-b][1,2,4]triazole-5-methanamine, 6-phenyl-α-(trifluoromethyl)-, (αR)-, TT-TFM) suppressed the activation of macrophages. Consistent with the in vitro data, TT-TFM protected against 2,4,6-trinitrobenzene sulfonic acid (TNBS), dextran sulfate sodium (DSS)-induced acute colitis and IL-10 knockout (KO) chronic colitis, as judged by body weight changes and colonic pathological damage. A mechanistic study based on microarray analysis and gene interference experiments indicated that TT-TFM exerted anti-inflammatory effects by enhancing Twist1 expression and subsequently blocking the NF-κB/TNF-α pathway. In addition, pretreatment with lentiviruses encoding shRNA targeting Twist1 could abolish the therapeutic effect of TT-TFM in TNBS colitis. Ultimately, TT-TFM showed anti-colitis activity by reducing NF-κB activation and the TNF-α level by promoting Twist1 expression; thus, TT-TFM may offer a therapeutic strategy for CD patients.
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Colitis/metabolismo , Activación de Macrófagos/efectos de los fármacos , Proteínas Nucleares/biosíntesis , Transducción de Señal/fisiología , Triazoles/química , Triazoles/uso terapéutico , Proteína 1 Relacionada con Twist/biosíntesis , Animales , Células Cultivadas , Colitis/tratamiento farmacológico , Femenino , Activación de Macrófagos/fisiología , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Endogámicos ICR , Ratones Noqueados , Proteínas Nucleares/agonistas , Transducción de Señal/efectos de los fármacos , Triazoles/farmacología , Trifluoroetanol/química , Trifluoroetanol/farmacología , Trifluoroetanol/uso terapéutico , Proteína 1 Relacionada con Twist/agonistasRESUMEN
The aim of this study was to prepare and characterize a novel type of starch-coated microparticles (MPs) allowing site-specific delivery of bioactives to the colon. An oral colon-specific controlled-release system was developed in the form of MPs coated with a resistant starch (RS2/RS3) film (RS@MPs) through an aqueous suspension coating process. The RS2 was chosen from a high-amylose cornstarch with 88.5% digestion resistibility. The RS3 was prepared by a high-temperature/pressure (HTP) treatment, with the following of enzymatic debranching, and retrogradation, resulting in a dramatic increase in enzymatic resistance (RS3 content: 76.6%). RS@MPs showed 40.7% of 5-aminosalicylic acid release within 8â¯h. The in vivo study of fluorescein-loaded RS@MPs indicated the high acidic and enzymatic resistibility of RS@MPs and a restrained release in the upper GIT. Therefore, RS@MPs has revealed to be a high potential system for accurately targeting bioactive compound delivery to the colon.
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Colon/metabolismo , Portadores de Fármacos/química , Microesferas , Almidón/química , Administración Oral , Animales , Liberación de Fármacos , Masculino , Mesalamina/administración & dosificación , Mesalamina/química , Mesalamina/metabolismo , Ratones , Especificidad de Órganos , Agua/químicaRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: The pharmacological effect derived from herb-herb interaction is important to constitute the prescription especially in traditional oriental medicine. The relationship of two medicinal herbs is called "couplet medicinals" which is used in pair for the purpose of enhancing the therapeutic effect, reducing the toxic effect or the adverse effect. The "Eighteen Incompatible Medicaments" constitute one of the contents in the incompatibility of traditional oriental drugs in a prescription. Among the "Eighteen Incompatible Medicaments", the roots and rhizomes of Veratrum nigrum (VN), is incompatible with the roots and rhizomes of Panax ginseng (PG). However, definite evidences of adverse effect by these combinations has yet to be reported. MATERIALS AND METHODS: The aim of the present study was to investigate the effects of ethanol extracts of PG, VN, and their combination (Pâ¯+â¯V) on the metastatic ability of colorectal cancer (CRC) cells using WST assay, flow cytometry, western blot analysis, real-time RT-PCR, immunofluorescence, migration assay, invasion assay, zymography, and an in vivo experiment with a lung-metastasis mouse model. RESULTS: The PG extract decreased cell proliferation by inducing cell cycle arrest and apoptosis of CRC cells. In addition, PG inhibited metastatic abilities of CRC cells including Epithelial-Mesenchymal Transition, migration, and invasion. Additionally, the PG extract suppressed lung metastasis of the CRC cells in the mouse model. However, the Pâ¯+â¯V extract exhibited weaker anti-proliferative and anti-metastatic effects than PG alone. CONCLUSION: Based on these results, the Pâ¯+â¯V couplet medicinal attenuates the anti-metastatic effects of PG, both in vitro and in vivo.
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Antineoplásicos Fitogénicos/farmacología , Panax/química , Extractos Vegetales/farmacología , Veratrum/química , Animales , Antineoplásicos Fitogénicos/administración & dosificación , Apoptosis/efectos de los fármacos , Puntos de Control del Ciclo Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Neoplasias Colorrectales/tratamiento farmacológico , Neoplasias Colorrectales/patología , Transición Epitelial-Mesenquimal/efectos de los fármacos , Femenino , Humanos , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/secundario , Ratones , Ratones Endogámicos BALB C , Extractos Vegetales/administración & dosificación , Raíces de Plantas , RizomaRESUMEN
Although metabolic reprogramming and redox imbalance are widely reported to be involved in chemo-resistance in cancer treatment, much more attention was paid to anti-cancer drug induced effect. Our previous studies showed that cancer cells can develop P-gp overexpression-mediated intrinsic drug resistance in the formation of 3D MCF-7 multi-cellular layers (MCLs) without any drug induction. However, whether metabolic reprogramming and redox imbalance functioned during this progress remained unrevealed. In our present study, LC-Q/TOF-MS and GC-MS were used in combination for analysing intracellular metabolites. The contribution of pentose phosphate pathway (PPP) and its related redox status were checked by chemical interfering and silencing/over-expression of glucose-6-phosphate dehydrogenase (G6PD). The downstream products of G6PD were assayed by quantitative real-time PCR, western blot and flow cytometry. Results showed that not only G6PD expression but also G6PD activity was significantly lowered along with 3D MCF-7 cells culture time. Impaired PPP disturbed redox-cycling, generated reactive oxygen species (ROS), which triggered cell cycle arrest and caused the switch to Chk2/p53/NF-κB pathway-mediated P-gp induction. Our results provided a new attempt to associate intrinsic small molecule metabolites (impaired PPP) communicating with cell signalling pathways through disturbed intracellular redox status to elucidate multi-cellular resistance (MCR) in 3D MCF-7 cells, which improved the understanding of the mechanisms of P-gp up-regulation in MCR with metabolomic and related redox status support.
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Subfamilia B de Transportador de Casetes de Unión a ATP/genética , Neoplasias de la Mama/genética , Glucosafosfato Deshidrogenasa/genética , Vía de Pentosa Fosfato/efectos de los fármacos , Antineoplásicos/farmacología , Neoplasias de la Mama/tratamiento farmacológico , Resistencia a Antineoplásicos/genética , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Glucosafosfato Deshidrogenasa/antagonistas & inhibidores , Humanos , Células MCF-7 , Metabolómica , FN-kappa B/genética , Oxidación-Reducción/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/efectos de los fármacos , Bibliotecas de Moléculas Pequeñas/farmacologíaRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Aconiti Sinomontani Radix is frequently used in the treatment of Bi syndrome in traditional Chinese medicine. Several reports indicate that Aconiti Sinomontani Radix has therapeutic effects for rheumatoid arthritis (RA). However, the cellular mode of action is still unclear. To investigate the effect of alkaloid extracts of Aconiti Sinomontani Radix on proliferation and migration of human synovial sarcoma SW982 cells as well as the molecular mechanism underlying. MATERIALS AND METHODS: SW982 cells were examined for proliferation by a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) method. Wound scratch assays were performed to assess the migrated rate of SW982 cells. Quantitative real-time PCR was used to measure the mRNA expression levels of Wnt5a, Runx2, MMP3, and Bmp2. Western blotting was used to measure the phosphorylated levels of JNK and NF-κB as well as the expression of MMP3. RESULTS: The alkaloid extract from Aconiti Sinomontani Radix (MQA) and MQB, which removed lappaconitine from MQA significantly inhibited the proliferation of SW982 in a dose-dependent manner. The proliferation inhibitory effect of MQB was more potent. Incubation with 10µg/ml MQB for 12, 24, and 36h inhibited the migration of SW982 cells by 83%, 58%, and 42%, respectively. Treatment with different concentrations of MQB for 24h inhibited mRNA expression of Wnt5a, Runx2, and MMP3, but Bmp2 mRNA expression was elevated by MQB. Further, MQB inhibited phosphorylation of JNK and NF-κB p65 as well as MMP3 expression by Western blotting analysis. CONCLUSION: The results showed that MQB inhibited proliferation and migration of SW982 cells possibly through suppressing Wnt5a-mediated JNK and NF-κB pathways. These results indicated that MQB might be an active extract of Aconiti Sinomontani Radix for targeting fibroblast-like synoviocytes (FLS) and be potential for RA therapy.
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Aconitum/química , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Extractos Vegetales/farmacología , Sinoviocitos/citología , Sinoviocitos/efectos de los fármacos , Proteína Morfogenética Ósea 2/biosíntesis , Línea Celular , Ensayos de Migración Celular , Subunidad alfa 1 del Factor de Unión al Sitio Principal/biosíntesis , Relación Dosis-Respuesta a Droga , Fibroblastos/citología , Expresión Génica/efectos de los fármacos , Humanos , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Metaloproteinasa 3 de la Matriz/biosíntesis , FN-kappa B/metabolismo , Fosforilación/efectos de los fármacos , Proteína Wnt-5a/biosíntesisRESUMEN
Olive oil vascular benefits have been attributed to hydroxytyrosol (HT). However, HT biological actions are still debated because it is extensively metabolized into glucuronides (GCs). The aim of this study was to test HT and GC vasculoprotective effects and the underlying mechanisms using aorta rings from 8-week-old male Wistar rats. In the absence of oxidative stress, incubation with 100 µM HT or GC for 5 min did not exert any vasorelaxing effect and did not influence the vascular function. Conversely, in condition of oxidative stress [upon incubation with 500 µM tert-butylhydroperoxide (t-BHP) for 30 min], preincubation with HT or GC improved acetylcholine-induced vasorelaxation compared with untreated samples (no t-BHP). This protective effect was lost for GC, but not for HT, when a washing step (15 min) was introduced between preincubation with HT or GC and t-BHP addition, suggesting that only HT enters the cells. In agreement, bilitranslocase inhibition with 100 µM phenylmethanesulfonyl fluoride for 20 min reduced significantly HT, but not GC, effect on the vascular function upon stress induction. Moreover, GC protective effect (improvement of endothelium-dependent relaxation in response to acetylcholine) in oxidative stress conditions was reduced by preincubation of aorta rings with 300 µM D-saccharolactone to inhibit ß-glucuronidase, which can deconjugate polyphenols. Finally, only HT was detected by high-pressure liquid chromatography in aorta rings incubated with GC and t-BHP. These results suggest that, in conditions of oxidative stress, GC can be deconjugated into HT that is transported through the cell membrane by bilitranslocase to protect vascular function.
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Antioxidantes/metabolismo , Ceruloplasmina/metabolismo , Endotelio Vascular/metabolismo , Glucuronidasa/metabolismo , Glucurónidos/metabolismo , Estrés Oxidativo , Alcohol Feniletílico/análogos & derivados , Animales , Antioxidantes/química , Aorta Torácica , Transporte Biológico Activo/efectos de los fármacos , Ceruloplasmina/antagonistas & inhibidores , Suplementos Dietéticos , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/enzimología , Inhibidores Enzimáticos/farmacología , Ácido Glucárico/análogos & derivados , Ácido Glucárico/farmacología , Glucuronidasa/antagonistas & inhibidores , Glucurónidos/química , Técnicas In Vitro , Masculino , Moduladores del Transporte de Membrana/farmacología , Oxidantes/farmacología , Estrés Oxidativo/efectos de los fármacos , Alcohol Feniletílico/química , Alcohol Feniletílico/metabolismo , Fluoruro de Fenilmetilsulfonilo/farmacología , Ratas Wistar , Enfermedades Vasculares/enzimología , Enfermedades Vasculares/metabolismo , Enfermedades Vasculares/prevención & control , Vasodilatación/efectos de los fármacos , terc-Butilhidroperóxido/farmacologíaRESUMEN
Our current knowledge of the implications of endocannabinoids in fear and anxiety is largely based on fear conditioning paradigms and approach-avoidance conflicts. Here we establish the ethobehavioral beetle mania task (BMT), which confronts mice with an erratically moving robo-beetle. With the help of this task we demonstrate decreased tolerance yet increased avoidance responses to an approaching beetle in high-anxiety behavior (HAB) and BALBc mice compared to C57BL/6N, CD1 and normal-anxiety behavior (NAB) mice. Also DBA/2N mice showed decreased passive and increased active behavior, but followed the robo-beetle more often than HAB and BALBc mice. Treatment with diazepam (1 mg/kg) increased tolerance without affecting avoidance behavior in HAB mice. Treatment with the MAGL inhibitor JZL184 (8 mg/kg) increased flight behavior, but did not affect tolerance. The FAAH inhibitor URB597 (0.3 mg/kg), however, reduced flight behavior and enhanced tolerance to the robo-beetle. The latter effects were blocked by co-treatment with the CB1 receptor antagonist SR141716A (3 mg/kg), which failed to affect the behavior by itself. Taken together, we validate the BMT as a novel test for studying endocannabinoids beyond traditional paradigms and for assessing active fear responses in mice. Furthermore, we demonstrate panicolytic consequences of pharmacological enhancement of anandamide, but not 2-AG signaling.