RESUMEN
Background: Hyperuricemia (HU) is a metabolic disease characterized by high uric acid levels in the blood. HU is a risk factor for diabetes, cardiovascular complications, metabolic syndrome, and chronic kidney disease. Purpose: The present study was performed to determine the effect of experimental HU on xanthine oxidase (XO), tumor necrosis factor-alpha (TNF-α), nuclear factor-kappa B (NF-κB), interleukin-17 (IL-17), cytochrome C, glutathione peroxidase (GPx), caspase-3, and 8-hydroxydeoxyguanosine (8-OHdG) levels in liver tissues of rats. Study Design: Thirty-five, male, Wistar albino-type rats were used for this study. Experimental groups were formed as follows: Group 1: control group; Group 2: potassium oxonate (PO) group; group 3: PO+NAR (naringenin; 2 weeks) group; and Group 4: PO (2 weeks)+NAR (2 weeks) group (total of 4 weeks). Methods: The first group was not given anything other than normal rat food and drinking water. In the second group, a 250 mg/kg intraperitoneal dose of PO was administered for 2 weeks. In the third group, 250 mg/kg intraperitoneal PO (application for 2 weeks) and 100 mg/kg NAR intraperitoneally 1 hr after each application were administered. In the fourth group, intraperitoneal PO administration was applied for 2 weeks, followed by intraperitoneal administration of NAR for 2 weeks (4 weeks in total). At the end of the experimental period, XO, TNF-α, NF-κB, IL-17, cytochrome C, GPx, caspase-3, and 8-OHdG levels were determined in liver tissues. Results: HU increased XO, TNF-α, NF-κB, IL-17, cytochrome C, caspase-3, and 8-OHdG levels in liver tissues. However, both 2 and 4 weeks of NAR supplementation decreased these values, and also NAR supplementation led to an increase in GPx levels in tissues. Conclusions: The results of the study show that increased inflammation, apoptosis, and DNA damage in experimental HU can be prevented by administration of NAR due to inhibition of cytochrome C, NF-κB, caspase-3, and 8-OHdG.
Asunto(s)
Agua Potable , Hiperuricemia , Masculino , Ratas , Animales , FN-kappa B/genética , FN-kappa B/metabolismo , FN-kappa B/farmacología , Caspasa 3/genética , Caspasa 3/metabolismo , Caspasa 3/farmacología , Interleucina-17/genética , Interleucina-17/metabolismo , Interleucina-17/farmacología , Factor de Necrosis Tumoral alfa/metabolismo , Citocromos c/genética , Citocromos c/metabolismo , Citocromos c/farmacología , 8-Hidroxi-2'-Desoxicoguanosina , Xantina Oxidasa/genética , Xantina Oxidasa/metabolismo , Xantina Oxidasa/farmacología , Ácido Úrico , Glutatión Peroxidasa/genética , Glutatión Peroxidasa/metabolismo , Glutatión Peroxidasa/farmacología , Agua Potable/efectos adversos , Agua Potable/metabolismo , Ratas Wistar , Apoptosis , Inflamación/metabolismo , Hígado/metabolismo , Daño del ADNRESUMEN
OBJECTIVE: To observe the effects of moxibustion at bilateral Feishu (BL13) and Xinshu (BL15) combined with benazepril on myocardial cells apoptosis index, the expression levels of apoptosis-related proteins cytochrome c (Cyt-C) and apoptosis-inducing factor (AIF) in chronic heart failure (CHF) rats. METHODS: Sixty-five rats were randomly divided into normal group () and model-I group (). After modeling, CHF rats in model-I group were divided into model group, moxibustion group, benazepril group, moxibustion plus benazepril group (abbreviated as aibei group, the same below), 10 rats in each group. Echocardiogram index was examined by echocardiography. Hemodynamic indices were measured by rat cardiac function meter. Serum B-type brain natriuretic peptide (BNP) was detected by enzyme-linked immunosorbent assay. Myocardial cells apoptosis index was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling staining. Pathological changes of myocardial tissues were observed by hematoxylin and eosin staining. The expression levels of Cyt-C and AIF in myocardial tissues were detected by Western blot. RESULTS: Compared with normal group, ejection fraction and left ventricular diameter shortening rate in model-â group were significantly reduced, myocardial cells of rats in model group exhibited unclear transverse striations, cells swellings and vacuoles, cardiac functions were deteriorated, serum BNP level, myocardial cells apoptosis index, and the expression levels of Cyt-C and AIF were significantly increased. Compared with model group, myocardial cells of rats in moxibustion group, benazepril group, and aibei group were dyed more evenly, muscle fibers were arranged relatively neatly, cardiac functions were improved, serum BNP level, myocardial cells apoptosis index, and the expression levels of Cyt-C and AIF were significantly decreased. Compared with aibei group, cardiac functions were worsened, myocardial cells apoptosis index, and the expression levels of Cyt-C and AIF were increased. CONCLUSION: Moxibustion at bilateral Feishu (BL13) and Xinshu (BL15) combined with benazepril could improve CHF better than moxibustion at bilateral Feishu (BL13) and Xinshu (BL15) or benazepril alone. The mechanisms might be that they can inhibit the expressions of Cyt-C and AIF, and inhibit the apoptosis of cardiomyocytes.
Asunto(s)
Insuficiencia Cardíaca , Moxibustión , Animales , Apoptosis , Factor Inductor de la Apoptosis/metabolismo , Factor Inductor de la Apoptosis/farmacología , Benzazepinas , Enfermedad Crónica , Citocromos c/genética , Citocromos c/metabolismo , Insuficiencia Cardíaca/tratamiento farmacológico , Insuficiencia Cardíaca/genética , Humanos , Ratas , Ratas Sprague-DawleyRESUMEN
In apoptotic pathway, the interaction of Cytochrome c (Cytc) with cardiolipin in vivo is a key process to induce peroxidase activity of Cytc and trigger the release of Cytc in the inner mitochondria into cytosol. The peroxidase active form of Cytc occurs due to local conformational changes that support the opening of the heme crevice and the loss of an axial ligand between Met80 and heme Fe. Structural adjustments at the Ω-loop segments of Cytc are required for such process. To study the role of the distal Ω-loop segments comprising residues 71-85 in human Cytc (hCytc), we investigated a cysteine mutation at Pro76, one of the highly conserved residues in this loop. The effect of P76C mutant was explored by the combination of experimental characterizations and molecular dynamics (MD) simulations. The peroxidase activity of the P76C mutant was found to be significantly increased by â¼13 folds relative to the wild type. Experimental data on global denaturation, alkaline transition, heme bleaching, and spin-labeling Electron Spin Resonance were in good agreement with the enhancement of peroxidase activity. The MD results of hCytc in the hexacoordinate form suggest the important changes in P76C mutant occurred due to the unfolding at the central Ω-loop (residues 40-57), and the weakening of H-bond between Tyr67 and Met80. Whereas the experimental data implied that the P76C mutant tend to be in equilibrium between the pentacoordinate and hexacoordinate forms, the MD and experimental information are complementary and were used to support the mechanisms of peroxidase active form of hCytc.
Asunto(s)
Citocromos c/metabolismo , Proteínas Mutantes/metabolismo , Peroxidasas/metabolismo , Secuencia de Aminoácidos , Cardiolipinas/metabolismo , Cisteína/química , Citocromos c/genética , Activación Enzimática , Hemo/metabolismo , Humanos , Simulación de Dinámica Molecular , Proteínas Mutantes/genética , Mutación , Conformación Proteica , Relación Estructura-ActividadRESUMEN
Anhydrosafflor yellow B (AHSYB) is a major active water-soluble pigment in Safflower, but it has not received enough attention yet. In this study, high-speed counter-current chromatography (HSCCC) was used to prepare AHSYB from safflower. The parameters of the separation process were optimized by response surface methodology for the first time. The entropy weight method (EWM) was applied to calculate the information entropy and the weight of five indexes, and then figure out a comprehensive index of the HSCCC separation effect. Under the optimized separation conditions, a HSCCC apparatus speed of 850 rpm, a flow rate of 2 mL min-1 for the mobile phase and a separation temperature of 40 °C for AHSYB were achieved with a purity of 98%. Furthermore, AHSYB was found to have cardio-protective effects by inhibiting apoptosis via the mitochondrial-mediated pathway in oxygen-glucose deprivation/reoxygenation-induced H9c2 cells. This research provides good method guides for the rapid and efficient separation of active compounds from food-grade Chinese herb medicines.
Asunto(s)
Apoptosis/efectos de los fármacos , Cardiotónicos/aislamiento & purificación , Cardiotónicos/farmacología , Carthamus tinctorius/química , Miocitos Cardíacos/efectos de los fármacos , Pigmentos Biológicos/aislamiento & purificación , Pigmentos Biológicos/farmacología , Adenosina Trifosfato/metabolismo , Animales , Cardiotónicos/química , Caspasa 3/genética , Caspasa 3/metabolismo , Forma de la Célula/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Distribución en Contracorriente , Citocromos c/genética , Citocromos c/metabolismo , Regulación hacia Abajo , L-Lactato Deshidrogenasa/metabolismo , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Mitocondrias Cardíacas/efectos de los fármacos , Mitocondrias Cardíacas/metabolismo , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/fisiología , Pigmentos Biológicos/química , Extractos Vegetales/química , Ratas , Especies Reactivas de OxígenoRESUMEN
Oxidative stress-induced cell damage and death of the retinal pigmented epithelium (RPE), a polarized monolayer that maintains retinal health and homeostasis, lead to the development of age-related macular degeneration (AMD). Several studies show that the naturally occurring antioxidant Lutein (Lut) can protect RPE cells from oxidative stress. However, the poor solubility and low oral bioavailability limit the potential of Lut as a therapeutic agent. In this study, lutein diglutaric acid (Lut-DG), a prodrug of Lut, was synthesized and its ability to protect human ARPE-19 cells from oxidative stress was tested compared to Lut. Both Lut and Lut-DG significantly decreased H2O2-induced reactive oxygen species (ROS) production and protected RPE cells from oxidative stress-induced death. Moreover, the immunoblotting analysis indicated that both drugs exerted their protective effects by modulating phosphorylated MAPKs (p38, ERK1/2 and SAPK/JNK) and downstream molecules Bax, Bcl-2 and Cytochrome c. In addition, the enzymatic antioxidants glutathione peroxidase (GPx) and catalase (CAT) and non-enzymatic antioxidant glutathione (GSH) were enhanced in cells treated with Lut and Lut-DG. In all cases, Lut-DG was more effective than its parent drug against oxidative stress-induced damage to RPE cells. These findings highlight Lut-DG as a more potent compound than Lut with the protective effects against oxidative stress in RPE cells through the modulation of key MAPKs, apoptotic and antioxidant molecular pathways.
Asunto(s)
Antioxidantes/farmacología , Células Epiteliales/efectos de los fármacos , Luteína/análogos & derivados , Estrés Oxidativo/efectos de los fármacos , Profármacos/farmacología , Epitelio Pigmentado de la Retina/efectos de los fármacos , Proteínas Reguladoras de la Apoptosis/biosíntesis , Proteínas Reguladoras de la Apoptosis/genética , Catalasa/biosíntesis , Catalasa/genética , Línea Celular , Citocromos c/biosíntesis , Citocromos c/genética , Evaluación Preclínica de Medicamentos , Células Epiteliales/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Glutatión/biosíntesis , Glutatión/genética , Glutatión Peroxidasa/biosíntesis , Glutatión Peroxidasa/genética , Humanos , Peróxido de Hidrógeno/toxicidad , Luteína/química , Luteína/farmacología , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Degeneración Macular/tratamiento farmacológico , Estructura Molecular , Especies Reactivas de Oxígeno/metabolismo , Epitelio Pigmentado de la Retina/citologíaRESUMEN
Neuronal cell death induced by chronic stress in the central nervous system is a cause of neurological dysfunction. We investigated the neuroprotective potential of a water extract of S. takesimense (WEST) against corticosterone-induced apoptosis in PC12 cells and the possible underlying mechanisms. Cells were pretreated with 50 µg/mL of WEST to evaluate its neuroprotective effect based on endoplasmic reticulum (ER) stress inhibition and mitochondrial function improvement. Pretreatment with WEST prevented corticosterone-induced injury in PC12 cells, resulting in increased cell survival, decreased lactate dehydrogenase (LDH) release, and potent apoptosis inhibition by a reduction in apoptotic nuclei demonstrated by Hoechst 33,342 and propidium iodide (PI) double staining, and TUNEL staining. WEST strongly attenuated calcium (Ca2+) elevation, inducing the closure of mitochondrial permeability transition pores (mPTPs), which were opened by corticosterone. It also stabilized mitochondrial membrane potential (MMP) loss and inhibited the corticosterone-induced decrease in adenosine triphosphate (ATP) levels. Furthermore, the increased reactive oxygen species (ROS) production induced by corticosterone was prevented in PC12 cells treated with WEST. WEST also downregulated the expression of glucose-regulated protein 78 (GRP78), growth arrest- and DNA damage-inducible gene 153 (GADD153), the pro-apoptotic protein Bcl-2-associated X (Bax), cytochrome c, cysteine-aspartic protease (caspase)-9, and caspase-3, and upregulated the expression of the anti-apoptotic protein B-cell lymphoma 2 (Bcl-2). Thus, WEST exerts a neuroprotective effect by inhibiting the apoptosis pathway in ER stress and the mitochondrial dysfunction induced by corticosterone. These results demonstrate that WEST reduces neuronal damage from the neurotoxicity caused by chronic stress.
Asunto(s)
Apoptosis/efectos de los fármacos , Corticosterona/efectos adversos , Fármacos Neuroprotectores/farmacología , Sedum/química , Adenosina Trifosfato/metabolismo , Animales , Caspasa 3/genética , Caspasa 3/metabolismo , Caspasa 9/genética , Caspasa 9/metabolismo , Supervivencia Celular/efectos de los fármacos , Citocromos c/genética , Citocromos c/metabolismo , Estrés del Retículo Endoplásmico/efectos de los fármacos , Regulación de la Expresión Génica , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , L-Lactato Deshidrogenasa/metabolismo , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Células PC12 , Extractos Vegetales/análisis , Extractos Vegetales/farmacología , Ratas , Especies Reactivas de Oxígeno/metabolismo , Factor de Transcripción CHOP/genética , Factor de Transcripción CHOP/metabolismo , Proteína X Asociada a bcl-2/genética , Proteína X Asociada a bcl-2/metabolismoRESUMEN
Autophagy is a universal self-degradation process involved in the removal and recycling of cellular constituents and organelles; however, little is known about its possible role in fruit ripening, in which the oxidation of lipids and proteins and changes in the metabolism of different cellular organelles occur. In this work, we analyzed several markers of autophagy in two critical maturation stages of pepper (Capsicum annuum L.) fruits where variations due to ripening become clearly visible. Using two commercial varieties that ripen to yellow and red fruits respectively, we studied changes in the gene expression and protein content of several autophagy (ATG) components, ATG4 activity, as well as the autophagy receptor NBR1 and the proteases LON1 and LON2. Additionally, the presence of intravacuolar vesicles was analyzed by electron microscopy. Altogether, our data reveal that autophagy plays a role in the metabolic changes which occur during ripening in the two studied varieties, suggesting that this process may be critical to acquiring final optimal quality of pepper fruits.
Asunto(s)
Autofagia , Capsicum/citología , Capsicum/crecimiento & desarrollo , Frutas/citología , Frutas/crecimiento & desarrollo , Biomarcadores/metabolismo , Capsicum/genética , Citocromos c/genética , Citocromos c/metabolismo , Frutas/genética , Regulación de la Expresión Génica de las Plantas , Humanos , Peroxidación de Lípido , Malato Sintasa/metabolismo , Estrés Oxidativo , Extractos Vegetales/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Vacuolas/metabolismo , Vacuolas/ultraestructuraRESUMEN
Theanine, an active component of green tea (Camellia sinensis), is considered a modulator of chemotherapy. To further investigate the anticancer activity of theanine, the present study investigated the cytotoxic effect of theanine at the concentration of 600 µg/ml, in the human HepG2 hepatoblastoma and HeLa adenocarcinoma cell lines, in comparison with the normal L02, H9c2 and HEK293 cell lines using a MTT assay. It was found that theanine induced cell death in the tumor cells, but not in the normal cells. Notably, when glutamine was restricted or reduced in the cell culture medium, the cell death induced by theanine was significantly enhanced. A terminal deoxynucleotidyltransferasemediated dUTP nick end labeling assay indicated that DNA damage was induced in theaninetreated HepG2 cells. Further experiments demonstrated that theanine caused HepG2 cell apoptosis through the mitochondrial pathway, with a loss of membrane potential and the release of apoptosisinducing factor, endonuclease G and cytochrome c. Western blot analysis and caspase activity detection also revealed that caspase9 and caspase3 were activated, whereas caspase8 remained inactive. These observations suggested that theanine exerted potent cytotoxicity on tumor cells when glutamine was restricted.
Asunto(s)
Apoptosis/efectos de los fármacos , Glutamatos/farmacología , Neoplasias/tratamiento farmacológico , Té/química , Caspasa 3/genética , Caspasa 8/genética , Proliferación Celular/efectos de los fármacos , Citocromos c/genética , Daño del ADN/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Glutamatos/química , Glutamina/genética , Glutamina/metabolismo , Células HEK293 , Células HeLa , Células Hep G2 , Humanos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Mitocondrias/efectos de los fármacos , Mitocondrias/genética , Transducción de Señal/efectos de los fármacosRESUMEN
NLRP3 inflammasome activation plays an important role in diabetic cardiomyopathy (DCM), which may relate to excessive production of reactive oxygen species (ROS). Gypenosides (Gps), the major ingredients of Gynostemma pentaphylla (Thunb.) Makino, have exerted the properties of anti-hyperglycaemia and anti-inflammation, but whether Gps improve myocardial damage and the mechanism remains unclear. Here, we found that high glucose (HG) induced myocardial damage by activating the NLRP3 inflammasome and then promoting IL-1ß and IL-18 secretion in H9C2 cells and NRVMs. Meanwhile, HG elevated the production of ROS, which was vital to NLRP3 inflammasome activation. Moreover, the ROS activated the NLRP3 inflammasome mainly by cytochrome c influx into the cytoplasm and binding to NLRP3. Inhibition of ROS and cytochrome c dramatically down-regulated NLRP3 inflammasome activation and improved the cardiomyocyte damage induced by HG, which was also detected in cells treated by Gps. Furthermore, Gps also reduced the levels of the C-reactive proteins (CRPs), IL-1ß and IL-18, inhibited NLRP3 inflammasome activation and consequently improved myocardial damage in vivo. These findings provide a mechanism that ROS induced by HG activates the NLRP3 inflammasome by cytochrome c binding to NLRP3 and that Gps may be potential and effective drugs for DCM via the inhibition of ROS-mediated NLRP3 inflammasome activation.
Asunto(s)
Antioxidantes/farmacología , Cardiotónicos/farmacología , Diabetes Mellitus Experimental/tratamiento farmacológico , Cardiomiopatías Diabéticas/tratamiento farmacológico , Inflamasomas/efectos de los fármacos , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Animales , Antioxidantes/aislamiento & purificación , Glucemia/efectos de los fármacos , Glucemia/metabolismo , Cardiotónicos/aislamiento & purificación , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Línea Celular , Citocromos c/genética , Citocromos c/metabolismo , Diabetes Mellitus Experimental/inducido químicamente , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/patología , Cardiomiopatías Diabéticas/inducido químicamente , Cardiomiopatías Diabéticas/genética , Cardiomiopatías Diabéticas/patología , Regulación de la Expresión Génica , Gynostemma/química , Inflamasomas/metabolismo , Interleucina-18/genética , Interleucina-18/metabolismo , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Masculino , Miocitos Cardíacos/citología , Miocitos Cardíacos/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/agonistas , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Estrés Oxidativo , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/farmacología , Ratas , Ratas Sprague-Dawley , Especies Reactivas de Oxígeno/antagonistas & inhibidores , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal , EstreptozocinaRESUMEN
As a natural compound, Ornithogalum caudatum Ait is primarily used as an anti-inflammatory and antitumor agent in Chinese folk medicine. In 1992, OSW-1 was isolated from this compound, which is a new member of cholestane saponin family. In numerous recent studies, OSW-1 has been shown to have powerful cytotoxic anticancer effects against various malignant cells. However, the therapeutic efficacy of OSW-1 on colon cancer and the underlying mechanism are not understood. To explore the mechanism underlying OSW-1 in antitumor therapy, a therapeutic function analysis of OSW-1 on colon cancer was performed in vitro and in vivo. It was shown that with low toxicity on normal colonic cells, OSW-1 suppresses colon cancer cells in vitro and this inhibition was via the intrinsic apoptotic pathway, which increased cellular calcium, changed mitochondrial membrane potential, disrupted mitochondrial morphology, and led to the release of cytochrome c and the activation of caspase-3. Furthermore, in a nude mouse model, OSW-1 had a powerful effect on suppressing colon tumor proliferation without significant side effects through the apoptosis pathway. Taken together, these results demonstrate that OSW-1 is a potential drug for colon cancer treatment.
Asunto(s)
Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Colestenonas/administración & dosificación , Neoplasias del Colon/tratamiento farmacológico , Saponinas/administración & dosificación , Animales , Caspasa 3/genética , Línea Celular Tumoral , Colestenonas/efectos adversos , Colestenonas/química , Neoplasias del Colon/genética , Neoplasias del Colon/patología , Citocromos c/genética , Citometría de Flujo , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Medicina Tradicional China , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Ratones , Saponinas/efectos adversos , Saponinas/química , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
BACKGROUND/AIMS: It has been shown, both experimentally and clinically, that water-pipe smoke (WPS) exposure adversely affects the cardiovascular system (CVS) through the generation of oxidative stress and inflammation. Betaine, a naturally occurring compound in common foods, has antioxidant and anti-inflammatory actions. However, its potential to mitigate the adverse effect of WPS on the CVS has never been reported before. This is the subject of this study in mice. METHODS: Mice were exposed daily for 30 min to either normal air (control), or to WPS for two consecutive weeks. Betaine was administered daily by gavage at a dose of 10mg/kg, 1h before either air or WPS exposure. RESULTS: Betaine mitigated the in vivo prothrombotic effect of WPS in pial arterioles and venules. Moreover, it reversed the WPS-induced decrease in circulating platelets. Likewise, betaine alleviated platelet aggregation in vitro, and the shortening of activated partial thromboplastin time and prothrombin time induced by WPS. Betaine reduced the increase of plasminogen activator inhibitor-1 and fibrinogen concentrations in plasma induced by WPS. Betaine also diminished the WPS-induced increase of plasma concentrations of interleukin 6 and tumor necrosis factor α, and attenuated the increase of lipid peroxidation and superoxide dismutase. Immunohistochemical analysis of the heart revealed an increase in the expression of inducible nitric oxide synthase and cytochrome C by cardiomyocytes of the WPS-exposed mice. These effects were averted by betaine. CONCLUSION: Our findings suggest that betaine treatment significantly mitigated WPS-induced hypercoagulability, and inflammation, as well as systemic and cardiac oxidative stress.
Asunto(s)
Antioxidantes/farmacología , Betaína/farmacología , Fumar/efectos adversos , Trombocitopenia/prevención & control , Trombosis/prevención & control , Administración Oral , Animales , Plaquetas/citología , Plaquetas/efectos de los fármacos , Plaquetas/metabolismo , Citocromos c/genética , Citocromos c/metabolismo , Fibrinógeno/genética , Fibrinógeno/metabolismo , Expresión Génica/efectos de los fármacos , Interleucina-6/genética , Interleucina-6/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Ratones , Ratones Endogámicos C57BL , Miocitos Cardíacos/citología , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Estrés Oxidativo/efectos de los fármacos , Tiempo de Tromboplastina Parcial , Inhibidor 1 de Activador Plasminogénico/genética , Inhibidor 1 de Activador Plasminogénico/metabolismo , Agregación Plaquetaria/efectos de los fármacos , Cultivo Primario de Células , Tiempo de Protrombina , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismo , Trombocitopenia/etiología , Trombocitopenia/metabolismo , Trombocitopenia/patología , Trombosis/etiología , Trombosis/metabolismo , Trombosis/patología , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
During the aging process, dimers of dietary vitamin A accumulated in retinal pigment epithelium (RPE) cells. Vitamin A dimer-mediated photooxidation resulted in RPE apoptosis, which is associated with age-related degenerative disease of retina, leading to blindness. It has been reported that proanthocyanidin-rich grape seed extract reduces oxidative stress in the eye. In this study, we investigated the underlying mechanism of photooxidation-induced apoptosis inhibition by procyanidins B2 (PB2), one of the main components of grape seed proanthocyanidin. To mimic vitamin A dimer-mediated photooxidation, ARPE-19 cells that accumulated vitamin A dimer, A2E, were used as a model system. Exposure of A2E loaded ARPE-19 cells to blue light induced ER stress and resulted in significant apoptosis. Pretreatment of blue light-exposed A2E containing ARPE-19 cells with PB2 inhibited apoptosis, increased the ratio of Bcl-2/Bax in the mitochondria, attenuated ROS and cytochrome c release, and decreased caspase cleavage. Additionally, PB2 inhibited the phosphorylation of ER stress markers elF2α and IRE1α and reduced CHOP expression. Moreover, PB2 inhibition of apoptosis is dependent on the UPR chaperone GRP78, indicating PB2 inhibits vitamin A dimer-mediated apoptosis in RPE cells by activating the UPR.
Asunto(s)
Apoptosis/efectos de los fármacos , Biflavonoides/farmacología , Catequina/farmacología , Estrés Oxidativo/efectos de los fármacos , Proantocianidinas/farmacología , Vitamina A/efectos adversos , Línea Celular , Citocromos c/genética , Citocromos c/metabolismo , Chaperón BiP del Retículo Endoplásmico , Estrés del Retículo Endoplásmico/efectos de los fármacos , Endorribonucleasas/genética , Endorribonucleasas/metabolismo , Silenciador del Gen , Extracto de Semillas de Uva/farmacología , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Humanos , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Fosforilación , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Retina/citología , Retina/efectos de los fármacos , Retina/metabolismo , Factor de Transcripción CHOP/genética , Factor de Transcripción CHOP/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Proteína X Asociada a bcl-2/genética , Proteína X Asociada a bcl-2/metabolismoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Acupuncture is a key part of traditional Chinese medicine, shown to induce favorable neuroplasticity for injuries in the central and peripheral nervous systems. Recent studies report elongated needle therapy (ENT) with BL54 and ST28 may restore acute spinal cord injury (ASCI). However, the precise mechanism for this has not been elucidated. AIM OF THE STUDY: In our current study, we investigated the effects of ENT on inflammation and neuronal apoptosis induced by ASCI, and whether PI3K/Akt and MAPK/ERK signaling pathways are involved in the ENT restoration effect. MATERIALS AND METHODS: Rat models of moderate SCI were established in accordance with the modified Allen's method and were treated with ENT continuously for 7 days. Spontaneous activities were evaluated by the Basso Beattie and Bresnahan locomotor scale. Levels of inflammatory cytokines, such as tumor necrosis factor alpha, interleukin-6, IL-1ß, and nuclear factor kappa-ß, were determined by enzyme-linked immunosorbent assay. Cell apoptosis was examined by the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The proportions of cells with positive Bcl-2 and Bax expression were determined by immunohistochemical assays, whilst the expression profiles of p-AKT and p-ERK in spinal cord tissues were evaluated by western blotting. Furthermore, the expression profiles of Cytochrome-C (Cyt-C) and caspase-3 in vivo were analyzed by reverse transcription polymerase chain reaction. The potential inhibitory effects downstream of the Akt and ERK signaling pathways were examined by administration of specific inhibitors LY294002 and PD98059 in vivo. RESULTS: As indicated by this study, inflammation as well as PI3K/Akt- and MAPK/ERK signaling pathway-mediated neuronal apoptosis were involved in the course of SCI in rats. The neuro-protective effect of ENT was associated with reduced Bax protein-positive neurons and increased Bcl-2 protein-positive neurons. ENT enhanced recovery of rat activities. Activation of p-Akt and p-ERK in the PI3K/Akt and MAPK/ERK signaling pathways, inhibited expression of the critical component Cyt-C. Cyt-C is required for the mitochondrial apoptosis pathway and cascade of caspase-3, which is involved in activation of neuronal apoptosis through down-regulation of Bax protein and up-regulation of Bcl-2, as determined by TUNEL. The administration of PI3K/Akt and MAPK/ERK signaling pathway specific inhibitors, LY294002 and PD98059, suppressed expression of both p-Akt and p-ERK. CONCLUSION: ENT with BL54 and ST28 points can promote the recovery of ASCI. And the neuro-protective effect of ENT during the restoration of SCI may be associated with the suppression of both inflammation and activation of PI3K/Akt and MAPK/ERK signaling pathways, resulting from down-regulation of Bax protein, up-regulation of Bcl-2, and inhibition of the mitochondrial apoptosis pathway.
Asunto(s)
Terapia por Acupuntura , Medicina Tradicional China , Agujas , Traumatismos de la Médula Espinal/terapia , Animales , Caspasa 3/genética , Cromonas/farmacología , Citocromos c/genética , Citocinas/metabolismo , Flavonoides/farmacología , Masculino , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Morfolinas/farmacología , FN-kappa B/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Proto-Oncogénicas c-akt/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Ratas Sprague-Dawley , Médula Espinal/metabolismo , Traumatismos de la Médula Espinal/metabolismo , Proteína X Asociada a bcl-2/metabolismoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Moringa oleifera Lam. is widely cultivated in Asian and African countries for its medicinal and dietary significance. The leaves are highly nutritious and are known to possess various biological activities. MATERIALS AND METHODS: Pre-pubertal Swiss albino male mice were injected with single dose of cyclophosphamide (CP, 200mg/kg body weight) or ethanolic extract of Moringa oleifera leaves (MOE, 100mg/kg body weight) intraperitoneally. In combination group, MOE was administered 24h prior to CP injection. RESULTS: CP induced a significant decrease in testicular weight (p<0.01) and depletion of germ cells (p<0.001) and higher level of DNA damage (p<0.001) compared to control. The expression of P53, Bax, Cytochrome C (Cyt C) was increased while there was a decrease in the expression of Bcl2, c-Kit and Oct4. Administration of MOE 24h prior to CP treatment ameliorated the depletion (p<0.001), DNA damage (p<0.001) and apoptosis (p<0.01) of germ cells induced by CP. The mitigating effect of MOE appears to be mediated by up-regulating the expression of c-Kit and Oct4 transcripts in P53-independent manner. CONCLUSION: MOE protects the spermatogonial cells from CP-induced damage by modulating the apoptotic response elicited by CP and therefore can be considered as an efficient method of male fertility preservation.
Asunto(s)
Ciclofosfamida/toxicidad , Moringa oleifera , Extractos Vegetales/farmacología , Sustancias Protectoras/farmacología , Espermatogonias/efectos de los fármacos , Animales , Citocromos c/genética , Daño del ADN , Etanol/química , Masculino , Ratones , Factor 3 de Transcripción de Unión a Octámeros/genética , Hojas de la Planta/química , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas c-kit/genética , ARN Mensajero/metabolismo , Desarrollo Sexual , Solventes/química , Recuento de Espermatozoides , Espermatogonias/metabolismo , Espermatogonias/patología , Proteína X Asociada a bcl-2/genéticaRESUMEN
PURPOSE: Wasabia japonica (wasabi) has been shown to exhibit properties of detoxification, anti-inflammation and the induction of apoptosis in cancer cells. This study aimed to investigate the molecular mechanism of the cytotoxicity of wasabi extract (WE) in colon cancer cells to evaluate the potential of wasabi as a functional food for chemoprevention. METHODS: Colo 205 cells were treated with different doses of WE, and the cytotoxicity was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide. Apoptosis and autophagy were detected by 4',6-diamidino-2-phenylindole, 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-imidacarbo-yanine iodide and staining for acidic vascular organelles (AVOs), along with Western blotting. RESULTS: The results demonstrated that WE induced the extrinsic pathway and mitochondrial death machinery through the activation of TNF-α, Fas-L, caspases, truncated Bid and cytochrome C. WE also induced autophagy by decreasing the phosphorylation of Akt and mTOR and promoting the expression of microtubule-associated protein 1 light chain 3-II and AVO formation. An in vivo xenograft model verified that tumor growth was delayed by WE treatment. CONCLUSION: Our studies revealed that WE exhibits anti-colon cancer properties through the induction of apoptosis and autophagy. These results provide support for the application of WE as a chemopreventive functional food and as a prospective treatment of colon cancer.
Asunto(s)
Apoptosis/efectos de los fármacos , Autofagia/efectos de los fármacos , Neoplasias del Colon/tratamiento farmacológico , Extractos Vegetales/farmacología , Wasabia/química , Animales , Antiinflamatorios/farmacología , Antineoplásicos/farmacología , Proteína Proapoptótica que Interacciona Mediante Dominios BH3/genética , Proteína Proapoptótica que Interacciona Mediante Dominios BH3/metabolismo , Línea Celular Tumoral/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Neoplasias del Colon/patología , Citocromos c/genética , Citocromos c/metabolismo , Humanos , Indoles/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Proteínas Asociadas a Microtúbulos/genética , Proteínas Asociadas a Microtúbulos/metabolismo , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Fosforilación , Proteínas Proto-Oncogénicas c-akt/genética , Proteínas Proto-Oncogénicas c-akt/metabolismo , Serina-Treonina Quinasas TOR/genética , Serina-Treonina Quinasas TOR/metabolismo , Sales de Tetrazolio/metabolismo , Tiazoles/metabolismo , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismo , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
This study describes a method for discriminating the true Cervus antlers from its counterfeits using multiplex PCR. Bioinformatics were carried out to design the specific alleles primers for mitochondrial (mt) cytochrome b (Cyt b) and cytochrome C oxidase subunit 1 (Cox 1) genes. The mt DNA and genomic DNA were extracted from Cervi Cornu Pantotrichum through the modified alkaline and the salt-extracting method in addition to its counterfeits, respectively. Sufficient DNA templates were extracted from all samples used in two methods, and joint fragments of 354 bp and 543 bp that were specifically amplified from both of true Cervus antlers served as a standard control. The data revealed that the multiplex PCR-based assays using two primer sets can be used for forensic and quantitative identification of original Cervus deer products from counterfeit antlers in a single step.
Asunto(s)
Cuernos de Venado , Citocromos b/genética , Citocromos c/genética , Ciervos/genética , Medicina Tradicional China/normas , Reacción en Cadena de la Polimerasa Multiplex , Oxidorreductasas/genética , Animales , ADN Mitocondrial , Genoma Mitocondrial , Reacción en Cadena de la Polimerasa Multiplex/métodosRESUMEN
The PI3K-Akt-mTOR, Wnt/ß-catenin and apoptosis signaling pathways have been shown to be involved in genesis of colorectal cancer (CRC). The aim of this study was to elucidate whether combination of Gelam honey and ginger might have chemopreventive properties in HT29 colon cancer cells by modulating the mTOR, Wnt/ß-catenin and apoptosis signaling pathways. Treatment with Gelam honey and ginger reduced the viability of the HT29 cells dose dependently with IC50 values of 88 mg/ml and 2.15 mg/ml respectively, their while the combined treatment of 2 mg/ml of ginger with 31 mg/ml of Gelam honey inhibited growth of most HT29 cells. Gelam honey, ginger and combination induced apoptosis in a dose dependent manner with the combined treatment exhibiting the highest apoptosis rate. The combined treatment downregulated the gene expressions of Akt, mTOR, Raptor, Rictor, ß-catenin, Gsk3ß, Tcf4 and cyclin D1 while cytochrome C and caspase 3 genes were shown to be upregulated. In conclusion, the combination of Gelam honey and ginger may serve as a potential therapy in the treatment of colorectal cancer through inhibiton of mTOR, Wnt/ß catenin signaling pathways and induction of apoptosis pathway.
Asunto(s)
Anticarcinógenos/farmacología , Miel , Extractos Vegetales/farmacología , Serina-Treonina Quinasas TOR/genética , Vía de Señalización Wnt/efectos de los fármacos , Zingiber officinale , Proteínas Adaptadoras Transductoras de Señales/genética , Apoptosis/efectos de los fármacos , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/genética , Proteínas Portadoras/genética , Caspasa 3/genética , Supervivencia Celular/efectos de los fármacos , Quimioprevención , Ciclina D1/genética , Citocromos c/genética , Relación Dosis-Respuesta a Droga , Regulación hacia Abajo/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Glucógeno Sintasa Quinasa 3/genética , Glucógeno Sintasa Quinasa 3 beta , Células HT29 , Humanos , Concentración 50 Inhibidora , Melaleuca , Proteínas Proto-Oncogénicas c-akt/genética , Proteína Asociada al mTOR Insensible a la Rapamicina , Proteína Reguladora Asociada a mTOR , Serina-Treonina Quinasas TOR/metabolismo , Factor de Transcripción 4 , Factores de Transcripción/genética , Regulación hacia Arriba/efectos de los fármacos , beta Catenina/genéticaRESUMEN
Naringin, the primary active compound of the traditional Chinese medicine Rhizoma drynariae, possesses many pharmacological activities. The present study is an effort to explore the anti-osteoporosis potential of naringin in vivo and in vitro. In vivo, we used ovariectomized rats to clarify the mechanisms by which naringin anti-osteoporosis. In vitro, we used osteoclasts to investigate naringin promotes osteoclasts apoptosis. Naringin was effective at enhancing BMD, trabecular thickness, bone mineralization, and mechanical strength in a dose-dependent manner. The result of RT-PCR analysis revealed that naringin down-regulated the mRNA expression levels of BCL-2 and up-regulated BAX, caspase-3 and cytochrome C. In addition, naringin significantly reduced the bone resorption area in vitro. These findings suggest that naringin promotes the apoptosis of osteoclasts by regulating the activity of the mitochondrial apoptosis pathway and prevents OVX-induced osteoporosis in rats.
Asunto(s)
Conservadores de la Densidad Ósea/farmacología , Resorción Ósea/prevención & control , Flavanonas/farmacología , Osteoclastos/efectos de los fármacos , Osteoporosis/prevención & control , Ovariectomía/efectos adversos , Animales , Apoptosis/efectos de los fármacos , Densidad Ósea/efectos de los fármacos , Conservadores de la Densidad Ósea/aislamiento & purificación , Resorción Ósea/genética , Resorción Ósea/metabolismo , Resorción Ósea/patología , Calcificación Fisiológica/efectos de los fármacos , Caspasa 3/genética , Caspasa 3/metabolismo , Línea Celular , Citocromos c/genética , Citocromos c/metabolismo , Relación Dosis-Respuesta a Droga , Femenino , Flavanonas/aislamiento & purificación , Expresión Génica/efectos de los fármacos , Humanos , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Osteoclastos/metabolismo , Osteoclastos/patología , Osteoporosis/etiología , Osteoporosis/genética , Osteoporosis/metabolismo , Extractos Vegetales/química , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
Isocudraxanthone K (IK) is a novel, natural compound from a methanol extract of the root bark of Cudrania tricuspidata. It has not been shown previously that IK possessed antitumor activity. We investigated the antitumor effects and molecular mechanism of IK and related signal transduction pathway(s) in oral squamous cell carcinoma cells (OSCCCs). The MTT assay revealed that IK had an antiproliferative effect on OSCCCs, in a dose- and time-dependent manner. IK induced apoptosis in OSCCCs, as identified by a cell-cycle analysis, annexin V-FITC and propidium iodide staining, and the nuclear morphology in cell death. IK caused time-dependent phosphorylation of Akt, p38, and ERK (extracellular signal-regulated kinase). In addition, IK increased the cytosolic to nuclear translocation of nuclear factor-κB (NF-κB) p65 and the degradation and phosphorylation of IκB-α in HN4 and HN12 cells. Furthermore, IK treatment downregulated hypoxia-inducible factor 1α (HIF-1α) and its target gene, vascular endothelial growth factor (VEGF). Cobalt chloride (CoCl2), a HIF-1α activator, attenuated the IK-induced growth-inhibiting and apoptosis-inducing effects, and blocked IK-induced expression of apoptosis regulatory proteins, such as Bax, Bcl-2, caspase-3, caspase-8, and caspase-9, and cytochrome c. Collectively, these data provide the first evidence of antiproliferative and apoptosis-inducing effects of IK as a HIF-1α inhibitor and suggest it may be a drug candidate for chemotherapy against oral cancer.
Asunto(s)
Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Moraceae/química , Neoplasias de la Boca , Corteza de la Planta/química , Extractos Vegetales/farmacología , Raíces de Plantas/química , Xantonas/farmacología , Caspasas/genética , Caspasas/metabolismo , Línea Celular Tumoral , Proliferación Celular/genética , Citocromos c/genética , Citocromos c/metabolismo , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Proteínas I-kappa B/genética , Proteínas I-kappa B/metabolismo , Neoplasias de la Boca/tratamiento farmacológico , Neoplasias de la Boca/genética , Neoplasias de la Boca/metabolismo , Neoplasias de la Boca/patología , Extractos Vegetales/química , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Factor de Transcripción ReIA/genética , Factor de Transcripción ReIA/metabolismo , Xantonas/química , Proteína X Asociada a bcl-2/genética , Proteína X Asociada a bcl-2/metabolismoRESUMEN
Rutin is derived from buckwheat, apples, and black tea. It has been shown to have beneficial anti-inflammatory and antioxidant effects. Ethanol is a central nervous system depressant and neurotoxin. Its metabolite, acetaldehyde, is critically toxic. Aldehyde dehydrogenase 2 (ALDH2) metabolizes acetaldehyde into nontoxic acetate. This study examined rutin's effects on ALDH2 activity in hippocampal neuronal cells (HT22 cells). Rutin's protective effects against acetaldehyde-based ethanol neurotoxicity were confirmed. Daidzin, an ALDH2 inhibitor, was used to clarify the mechanisms of rutin's protective effects. Cell viability was significantly increased after rutin treatment. Rutin significantly reversed ethanol-increased Bax, cytochrome c expression and caspase 3 activity, and decreased Bcl-2 and Bcl-xL protein expression in HT22 cells. Interestingly, rutin increased ALDH2 expression, while daidzin reversed this beneficial effect. Thus, this study demonstrates rutin protects HT22 cells against ethanol-induced neurotoxicity by increasing ALDH2 activity.