RESUMEN
The oral consumption of capsicum has been reported to increase interleukin (IL)-2 and interferon (IFN)-γ production in Peyer's patches (PP); however, the active components responsible for these effects have not been completely identified. The beneficial biological effects of green peppers cultivated under environmentally friendly farming conditions (ECP), without the use of chemical pesticides, have rarely been compared with those of green peppers cultivated under conventional farming conditions (CCP). Oral administration of ECP extract significantly induced the production of IL-2 and IFN-γ in concanavalin A-treated cells from PP ex vivo; their levels were much higher than those in the CCP extract-treated group. A comparative analysis of the HPLC profiles indicated a 1.7-fold increase of a peak, named EF-1, at 415 nm in the ECP extract. The major component of EF-1 was identified as pheophytin a, which is a chlorophyll a molecule lacking a central Mg(2+) ion, as determined from NMR data. Intake of pheophytin a and chlorophyll a significantly increased IL-2 and IFN-γ production, and the percentage of IL-2- and IFN-γ-producing CD4+ T-cells in PP. Taken together, our data suggest that ECPs produce a higher content of pheophytin a than CCPs, and pheophytin a and chlorophyll a are immune-modulating components in green vegetables.
Asunto(s)
Capsicum , Clorofila/farmacología , Interferón gamma/metabolismo , Interleucina-2/metabolismo , Ganglios Linfáticos Agregados/efectos de los fármacos , Feofitinas/farmacología , Agricultura/métodos , Animales , Linfocitos T CD4-Positivos/efectos de los fármacos , Linfocitos T CD4-Positivos/metabolismo , Células Cultivadas , Clorofila/aislamiento & purificación , Clorofila A , Masculino , Ratones , Ratones Endogámicos C57BL , Ganglios Linfáticos Agregados/metabolismo , Feofitinas/aislamiento & purificación , Extractos Vegetales/químicaRESUMEN
BACKGROUND: Bisphenol A (BPA), an endocrine disrupting chemical, has been suspected to pose carcinogenic risks. However, likely mechanisms are obscure and there are difficulties to estimating its real significance for cancer development. METHODS: We therefore studied BPA-induced proteomic alterations in immune organs of ICR mice offspring that were prenatally exposed to BPA (15 and 300 mg/L of drinking water). We performed 2D-gel analyses of samples, considering differences in spleen, exposure levels, sex, and ages. RESULTS: From proteomic analyses, we found various proteins were up- or down-regulated by BPA. Among them, SET, a putative oncogene and inhibitor of phosphatase 2A, was significantly down- regulated in a BPA dose-dependent manner. We also confirmed down-regulation of SET in western blot and real time PCR analyses. From gene network analysis, SET is predicted to communicate with other genes including CYP17, which is involved in biosynthesis and metabolism of sex-hormones. CONCLUSIONS: This study provided evidence that SET can be applied as a new biomarker for prenatal BPA exposure and suggests a potential new mechanism of action in that BPA may disrupt CYP17 via SET.
Asunto(s)
Compuestos de Bencidrilo/toxicidad , Disruptores Endocrinos/toxicidad , Proteínas Oncogénicas/genética , Proteínas Oncogénicas/metabolismo , Fenoles/toxicidad , Efectos Tardíos de la Exposición Prenatal , Esteroide 17-alfa-Hidroxilasa/genética , Animales , Biomarcadores , Proteínas de Unión al ADN , Regulación hacia Abajo , Femenino , Hormonas Esteroides Gonadales/biosíntesis , Hormonas Esteroides Gonadales/metabolismo , Chaperonas de Histonas , Masculino , Ratones , Ratones Endogámicos ICR , Embarazo , Complicaciones del Embarazo , Proteómica , Bazo/fisiología , Esteroide 17-alfa-Hidroxilasa/metabolismoRESUMEN
We investigated the comparative effects of 4 and 60 Hz magnetic fields on pentylenetetrazole (PTZ)-induced seizure in mice. For this study, we measured the latent time to seizure, seizure duration, and lethality induced by PTZ in mice exposed to 4 and 60 Hz magnetic fields (MF) for 30 min. Compared to sham-exposed controls, the latent time to tail twitching and seizure in the 4 Hz MF group was significantly decreased while the latent time to seizure in the 60 Hz MF group was significantly increased. The seizure duration in the 4 Hz MF group was significantly decreased while that in the 60 Hz MF group was significantly increased. More importantly, while the mice exposed to a 60 Hz MF experienced significantly increased lethality after seizure convulsion, those exposed to a 4 Hz MF showed no lethality, with a shortening of the duration of seizure. This beneficial effect of a 4 Hz MF on seizure has the same implication as the anti-oxidative effects of a 4 Hz MF observed in our previous work. The results of our current and previous works indicate that a 4 Hz MF may be used as a therapeutic physical agent for the treatment of oxidative stress-induced diseases, including seizure, with or without chemical drugs.
Asunto(s)
Magnetoterapia/métodos , Pentilenotetrazol/farmacología , Convulsiones/inducido químicamente , Convulsiones/terapia , Animales , Ratones , Estrés Oxidativo/efectos de los fármacos , Convulsiones/metabolismoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Chaga mushroom (Inonotus obliquus, IO) has been used as a folk remedy for cancer, digestive system diseases, and other illnesses in Russia and Eastern Europe. AIM OF THE STUDY: In the present study, we investigated the immunomodulating effects of IO through in vivo and ex vivo studies. MATERIALS AND METHODS: Serum immunoglobulins (IgE, IgG(1), and IgG(2a)) and cytokines (interleukin (IL)-4, interferon (IFN)-γ, and IL-2) were measured in concanavalin A (ConA)-stimulated splenocytes and CD4(+) T cells. The nitric oxide (NO) secretion of lipopolysaccharide (LPS)-stimulated peritoneal macrophages was also measured after oral administration of 50, 100, or 200 mg kg(-1) d(-1) IO hot water extract (IOE) to ovalbumin (OVA)-sensitized BALB/c mice. RESULTS: We found that the OVA-induced increase in serum IgE and IgG(2a) was significantly suppressed when IOE was orally administered after the second immunization with OVA. ConA stimulation in spleen cells isolated from OVA-sensitized mice treated with 100 mg kg(-1) IOE resulted in a 25.2% decrease in IL-4 production and a 102.4% increase in IFN-γ, compared to the controls. Moreover, IL-4, IFN-γ, and IL-2 were significantly reduced after ConA stimulation in isolated CD4(+)T cells. We also determined that IOE inhibits the secretion of NO from LPS-stimulated peritoneal macrophages ex vivo. CONCLUSIONS: We suggest that IO modulates immune responses through secretion of Th1/Th2 cytokines in immune cells and regulates antigen-specific antibody production.
Asunto(s)
Agaricales , Antígenos , Citocinas/metabolismo , Inmunoglobulina E/metabolismo , Factores Inmunológicos/farmacología , Ovalbúmina/inmunología , Bazo/efectos de los fármacos , Células TH1/efectos de los fármacos , Células Th2/efectos de los fármacos , Administración Oral , Agaricales/química , Animales , Células Cultivadas , Relación Dosis-Respuesta a Droga , Femenino , Inmunoglobulina G/metabolismo , Factores Inmunológicos/administración & dosificación , Factores Inmunológicos/aislamiento & purificación , Inyecciones Intraperitoneales , Interferón gamma/metabolismo , Interleucina-2/metabolismo , Interleucina-4/metabolismo , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/inmunología , Ratones , Ratones Endogámicos BALB C , Óxido Nítrico/metabolismo , Ovalbúmina/administración & dosificación , Ganglios Linfáticos Agregados/efectos de los fármacos , Ganglios Linfáticos Agregados/inmunología , Bazo/citología , Bazo/inmunología , Células TH1/inmunología , Células Th2/inmunologíaRESUMEN
The proliferation and migration of vascular smooth muscle cells (VSMCs) play important roles in the formation and progression of intimal thickening in early-phase atherosclerosis and in restenosis after vascular injury. Tumor necrosis factor-α (TNF-α) is released from macrophages in atherosclerotic lesions and from neointimal vascular smooth muscle cells after balloon-injury. Obovatol, a major biphenolic component isolated from the Magnolia obovata leaf, is known to have anti-inflammatory and antitumor activities. The goal of this study was to examine the cardioprotective effects of the obovatol derivative OD 78 on the TNF-α-induced proliferation and migration of rat aortic smooth muscle cells (RASMCs). The antiproliferative effects of OD 78 on RASMCs were examined by cell counting and [(3)H]-thymidine incorporation assays. Treatment of cells with 1-4 µM OD 78 inhibited the proliferation and DNA synthesis of TNF-α-stimulated RASMCs in a concentration-dependent manner, without cytotoxicity. Treatment with OD 78 inhibited TNF-α-mediated p38 phosphorylation, but did not change the activation of extracellular signal-regulated kinase or c-Jun N-terminal kinase. Furthermore, treatment with OD 78 decreased TNF-α-induced levels of cyclin E, cyclin D1, CDK2, proliferating cell nuclear antigen, and phosphorylated retinoblastoma protein, but not the CDK4 expression level. Also, OD 78 inhibits the migration of TNF-α-induced RASMC in transwells. OD 78 treatment strongly decreased matrix metalloproteinase-9 (MMP-9) expression in a dose-dependent manner, but the MMP-2 expression was unchanged. These results show that OD 78 may be developed as a potential antiproliferative agent for the treatment of angioplasty restenosis and atherosclerosis.
Asunto(s)
Benzoatos/farmacología , Cardiotónicos/farmacología , Proliferación Celular/efectos de los fármacos , Músculo Liso Vascular/efectos de los fármacos , Miocitos del Músculo Liso/efectos de los fármacos , Éteres Fenílicos/farmacología , Animales , Aorta/citología , Aorta/efectos de los fármacos , Benzoatos/metabolismo , Cardiotónicos/metabolismo , Ciclo Celular/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Ciclina E/metabolismo , ADN/análisis , ADN/biosíntesis , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Músculo Liso Vascular/citología , Músculo Liso Vascular/metabolismo , Miocitos del Músculo Liso/fisiología , Éteres Fenílicos/metabolismo , Fosforilación/efectos de los fármacos , Ratas , Proteína de Retinoblastoma/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
The effects of repeated hair washing and a single hair dyeing on concentrations of methamphetamine (MA) and amphetamine (AM) in hair samples of MA addicts were studied. Thirty-one MA positive hair samples collected from male (n = 24, 24-51 yrs) and female abusers (n = 7, 17-46 yrs) were evaluated for MA and AM concentration's changes after repeated hair washing and a single hair dyeing. Thirty-one MA positive hair samples, no additional treatment hair sample group (NAT), were treated in vitro with liquid soap or three kinds of hair dyes which were black, brown and yellow color hair dye, respectively. Quantitation of AM and MA in hair samples was utilized GC-MS using selected ion monitoring. MA and AM concentrations in NAT were 10.41 ± 8.91 ng/mg (range 1.50-30.0 ng/mg) and 2.24 ± 2.75 ng/mg (range 0.41-12.90 ng/mg). And, their concentrations were decreased about 23.3 ± 4.5% (range 16.7-32.8%) in hair repeated washing group (WAS) and 32.6 ± 4.82 (22.2-41.9) in three kinds of a single hair dyeing groups in comparison to original concentrations of MA and AM in NAT. A statistically significant difference was found between NAT and WAS or three hair dyeing groups (p < 0.01), but not between WAS and three hair dyeing groups, and not between each hair dyeing groups with each three kinds of hair dyes (p > 0.05).
Asunto(s)
Anfetamina/análisis , Estimulantes del Sistema Nervioso Central/análisis , Preparaciones para el Cabello , Cabello/química , Metanfetamina/análisis , Adulto , Femenino , Toxicología Forense , Cromatografía de Gases y Espectrometría de Masas , Humanos , Masculino , Persona de Mediana Edad , Detección de Abuso de Sustancias/métodos , Adulto JovenRESUMEN
Vascular smooth muscle cells (VSMCs) are the main cellular component in the arterial wall, and abnormal proliferation of VSMCs plays a central role in the pathogenesis of atherosclerosis and restenosis after angioplasty, and possibly in the development of hypertension. Pterostilbene, a natural dimethylated analog of resveratrol, is known to have diverse pharmacological activities including anti-cancer, anti-inflammation and anti-oxidant activities. The present study was designed to investigate the effects of pterostilbene on platelet-derived growth factor (PDGF)-BB-induced VSMCs proliferation as well as the molecular mechanisms of the antiproliferative effects. The cell growth of VSMCs was determined by cell counting and [(3)H]thymidine incorporation assays. Pterostilbene significantly inhibited the DNA synthesis and proliferation of PDGF-BB-stimulated VSMCs in a concentration-dependent manner. The inhibition percentages of pterostilbene at 1, 3 and 5microM to VSMCs proliferation were 68.5, 80.7 and 94.6%, respectively. The DNA synthesis of pterostilbene at 1, 3 and 5microM in VSMCs was inhibited by 47.4, 76.7 and 100%, respectively. Pterostilbene inhibited the PDGF-BB-stimulated phosphorylation of Akt kinase. However, pterostilbene did not change the expression of extracellular signal-related kinase (ERK) 1/2, PLCgamma1, phosphatidylinositol (PI)3 kinase and PDGF-Rbeta phosphorylation. In addition, pterostilbene down-regulated the cell cycle-related proteins including the expression of cyclin-dependent kinase (CDK) 2, cyclin E, CDK4, cyclin D1, retinoblastoma (Rb) proteins and proliferative cell nuclear antigen (PCNA). These findings suggest that the inhibition of pterostilbene to the cell proliferation and DNA synthesis of PDGF-BB-stimulated VSMCs may be mediated by the suppression of Akt kinase. Furthermore, pterostilbene may be a potential anti-proliferative agent for the treatment of atherosclerosis and angioplasty restenosis.
Asunto(s)
Músculo Liso Vascular/efectos de los fármacos , Miocitos del Músculo Liso/efectos de los fármacos , Proteínas Proto-Oncogénicas c-akt/efectos de los fármacos , Estilbenos/farmacología , Animales , Aorta/efectos de los fármacos , Aorta/metabolismo , Becaplermina , Proliferación Celular/efectos de los fármacos , Células Cultivadas , ADN/biosíntesis , ADN/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Músculo Liso Vascular/metabolismo , Miocitos del Músculo Liso/metabolismo , Fosforilación/efectos de los fármacos , Factor de Crecimiento Derivado de Plaquetas/farmacología , Proteínas Proto-Oncogénicas c-akt/metabolismo , Proteínas Proto-Oncogénicas c-sis , Ratas , Estilbenos/administración & dosificaciónRESUMEN
Fenofibrate, a lipid-lowering drug, inhibits hydroxyl-methylglutaryl coenzyme A (HMG-CoA)-reductase activity, thus reducing cholesterol synthesis and increasing the clearance of circulating LDL-cholesterol via the high affinity receptor system. In addition, fenofibrate has beneficial effects such as the inhibition of tissue factor expression, antithrombotic effect and anti-inflammatory effect. The aim of this study was to investigate the effects of fenofibrate on thrombus formation in vivo and platelet activation in vitro and ex vivo. The carotid arteries of male Sprague-Dawley rats were subjected to chemical injury by FeCl(3), and then blood flow was measured with a blood flowmeter. Fenofibrate (200 and 400mg/kg/day for 1 week) delayed the time to occlusion by 61.3% (p<0.05, n=10) and 90.7% (p<0.01, n=10), respectively. Fenofibrate also significantly inhibited ex vivo platelet aggregations induced by collagen (7.5microg/ml) (p<0.01, n=11) and ADP (10microM) (p<0.01, n=11), respectively, but did not affect coagulation times following activated partial thromboplastin and prothrombin activation, indicating the antithrombotic effect was mediated by its inhibition on platelet activation rather than coagulation system. This antiplatelet activity was revealed to be mediated by the suppression of thromboxane A(2) receptor, cytosolic calcium mobilization, and cyclooxygenase (COX)-1 activity. Taken together, we demonstrate that fenofibrate can significantly inhibit artery thrombus formation in vivo, which may be due to antiplatelet activity via the inhibition of thromboxane A(2) receptor, cytosolic calcium mobilization and COX-1 activity, and the beneficial effect of fenofibrate on cardiovascular system may be also due to its modulation of platelet activation.
Asunto(s)
Fenofibrato/farmacología , Inhibidores de Agregación Plaquetaria/farmacología , Agregación Plaquetaria/efectos de los fármacos , Trombosis/tratamiento farmacológico , Animales , Ácido Araquidónico/metabolismo , Coagulación Sanguínea/efectos de los fármacos , Calcio/metabolismo , Ciclooxigenasa 1/metabolismo , Fenofibrato/uso terapéutico , Fibrinolíticos/farmacología , Masculino , Conejos , Ratas , Ratas Sprague-Dawley , Serotonina/metabolismoRESUMEN
The interstitial cells of Cajal (ICC) are pacemaking cells required for gastrointestinal motility. The possibility of whether DA-9701, a novel prokinetic agent formulated with Pharbitis Semen and Corydalis Tuber, modulates pacemaker activities in the ICC was tested using the whole cell patch clamp technique. DA-9701 produced membrane depolarization and increased tonic inward pacemaker currents in the voltage-clamp mode. The application of flufenamic acid, a non-selective cation channel blocker, but not niflumic acid, abolished the generation of pacemaker currents induced by DA-9701. Pretreatment with a Ca2+-free solution and thapsigargin, a Ca2+-ATPase inhibitor in the endoplasmic reticulum, abolished the generation of pacemaker currents. In addition, the tonic inward currents were inhibited by U-73122, an active phospholipase C inhibitor, but not by GDP-beta-S, which permanently binds G-binding proteins. Furthermore, the protein kinase C inhibitors, chelerythrine and calphostin C, did not block the DA-9701-induced pacemaker currents. These results suggest that DA-9701 might affect gastrointestinal motility by the modulation of pacemaker activity in the ICC, and the activation is associated with the non-selective cationic channels via external Ca2+ influx, phospholipase C activation, and Ca2+ release from internal storage in a G protein-independent and protein kinase C-independent manner.
Asunto(s)
Intestino Delgado/efectos de los fármacos , Preparaciones de Plantas/farmacología , Fosfolipasas de Tipo C/antagonistas & inhibidores , Animales , Calcio/metabolismo , Femenino , Ácido Flufenámico/farmacología , Fármacos Gastrointestinales/farmacología , Motilidad Gastrointestinal/fisiología , Intestino Delgado/citología , Intestino Delgado/metabolismo , Masculino , Potenciales de la Membrana/efectos de los fármacos , Ratones , Ratones Endogámicos BALB C , Ácido Niflúmico/farmacología , Técnicas de Placa-Clamp , Fosfolipasas de Tipo C/metabolismoRESUMEN
Cudrania tricuspidata has been proposed to possess anti-inflammatory, antioxidant, hepatoprotective, and antitumor activities. Although cudraflavone B, isolated from the root bark of C. tricuspidata, has a variety of pharmacological effects, its effects on rat aortic smooth muscle cells (RASMCs) are unclear. In the present study, cudraflavone B was found to inhibit cell proliferation and DNA synthesis in cultured RASMCs. Pretreatment with cudraflavone B (0.1-4 microM) suppressed platelet-derived growth factor-BB (PDGF-BB)-stimulated cell number in a concentration-dependent manner. The inhibition percentages were 19.7%, 36.4%, 52.3%, and 99.1% at concentrations of 0.1, 1, 2, and 4 microM, respectively. Moreover, cudraflavone B inhibited [H]-thymidine incorporation into DNA in RASMCs in response to 25 ng/mL PDGF-BB. PDGF-BB-stimulated DNA synthesis was significantly reduced by 15.9%, 31.7%, 43.1%, and 78.2% at concentrations of 0.1, 1, 2, and 4 muM, respectively. Thus, cudraflavone B blocked the PDGF-BB-inducible progression through G0/G1 to S phase of the cell cycle in synchronized cells. Furthermore, PDGF-BB-induced phosphorylation of retinoblastoma protein (pRb), the hyperphosphorylation of which is a hallmark of the G1-S transition in the cell cycle, was significantly inhibited by cudraflavone B. Because pRb phosphorylation is regulated by cyclin-dependent kinases (CDKs), we investigated the expression of CDK2, CDK4, cyclin E, and cyclin D1 and the CDK inhibitors p21 and p27. Treatment with cudraflavone B downregulated the cyclins and CDKs and upregulated the expression of p21 and p27, a CDK inhibitor. These findings suggest that cudraflavone B inhibits RASMC proliferation via the induction of p21 and p27 expression and subsequent cell cycle arrest with reduction of pRb phosphorylation at the G1-S phase.
Asunto(s)
Proliferación Celular/efectos de los fármacos , Regulación hacia Abajo/efectos de los fármacos , Flavonoides/farmacología , Moraceae/química , Miocitos del Músculo Liso/efectos de los fármacos , Proteína de Retinoblastoma/metabolismo , Transducción de Señal/efectos de los fármacos , Animales , Aorta/citología , Células Cultivadas , Medio de Cultivo Libre de Suero , Relación Dosis-Respuesta a Droga , Flavonoides/química , Flavonoides/aislamiento & purificación , Estructura Molecular , Fosforilación/efectos de los fármacos , Corteza de la Planta/química , Raíces de Plantas/química , Factor de Crecimiento Derivado de Plaquetas/farmacología , Ratas , Factores de TiempoRESUMEN
Beta-carboline alkaloids including harmalol, harmaline, norharmane, harmol, harmine and harmane are important constituents of the medicinal plant, Perganum harmala L. (Zygophylaceae), which has been used in traditional medicine. In the present study, the antiplatelet activities of six beta-carboline alkaloid compounds were investigated in vitro. At a concentration of 200 microM, these compounds have no effect on arachidonic acid (AA)-, thrombin- and U46619 (a thromboxane A2 mimic)-stimulated platelet aggregation. On the contrary, it was revealed that collagen-induced platelet aggregation could be inhibited by these compounds with different potencies (harmane and harmine were most potent, harmol had medium potency, and harmol, norharmane, harmalol and harmaline had a weak, non significant effect), indicating a selective inhibition on collagen-mediated platelet activation. Consistently, further study revealed that collagen-mediated phospholipase (PL) Cgamma2 and protein tyrosine phosphorylation, cytosolic calcium mobilization and arachidonic acid liberation were completely inhibited by harmane and harmine in a concentration-dependent manner, while the other compounds were only partially or not effective at all. Taken together, these results indicate that three of these six beta-carboline alkaloids can selectively affect collagen-induced platelet aggregation with different potencies; in particular, harmane and harmine were most potent, and their antiplatelet activities may be mediated by inhibiting PLCgamma2 and protein tyrosine phosphorylation with sequential suppression of cytosolic calcium mobilization and arachidonic acid liberation, indicating that harmane and harmine have a potential to be developed as a novel agent for atherothrombotic diseases.
Asunto(s)
Alcaloides/farmacología , Carbolinas/farmacología , Peganum , Fosfolipasa C gamma/metabolismo , Agregación Plaquetaria/efectos de los fármacos , Animales , Ácido Araquidónico/metabolismo , Plaquetas/metabolismo , Calcio/metabolismo , Carbolinas/química , Técnicas In Vitro , Masculino , Estructura Molecular , Fosforilación/efectos de los fármacos , Fosfotirosina/metabolismo , Extractos Vegetales/farmacología , Inhibidores de Agregación Plaquetaria/farmacología , Conejos , Relación Estructura-ActividadRESUMEN
The proliferation of vascular smooth muscle cells (VSMCs) induced by injury to the intima of arteries is an important etiologic factor in vascular proliferative disorders such as atherosclerosis and restenosis. Uncaria rhynchophylla is traditional Chinese herb that has been applied to the treatment of convulsive disorders, such as epilepsy, in China. In the present study, we examined whether corynoxeine exerts inhibitory effects on platelet-derived growth factor (PDGF)-BB-induced rat aortic VSMC proliferation and the possible mechanism of such effects. Pre-treatment of VSMCs with corynoxeine (5-50 microM) for 24 h resulted in significant decreases in cell number without any cytotoxicity; the inhibition percentages were 25.0+/-12.5, 63.0+/-27.5 and 88.0+/-12.5% at 5, 20 and 50 microM, respectively. Also, corynoxeine significantly inhibited the 50 ng/ml PDGF-BB-induced DNA synthesis of VSMCs in a concentration-dependent manner without any cytotoxicity; the inhibitions were 32.8+/-11.0, 51.8+/-8.0 and 76.9+/-7.4% at concentrations of 5, 20 and 50 microM, respectively. Pre-incubation of VSMCs with corynoxeine significantly inhibited PDGF-BB-induced extracellular signal-regulated kinase 1/2 (ERK1/2) activation, whereas corynoxeine had no effects on mitogen-activated protein kinase (MAPK/ERK)-activating kinase 1 and 2 (MEK1/2), Akt, or phospholipase C (PLC)gamma1 activation or on PDGF receptor beta (PDGF-Rbeta) phosphorylation. These results suggest that corynoxeine is a potent ERK1/2 inhibitor of key PDGF-BB-induced VSMC proliferation and may be useful in the prevention and treatment of vascular diseases and restenosis after angioplasty.
Asunto(s)
Alcaloides/farmacología , Proliferación Celular/efectos de los fármacos , Medicamentos Herbarios Chinos/farmacología , Proteína Quinasa 1 Activada por Mitógenos/antagonistas & inhibidores , Proteína Quinasa 3 Activada por Mitógenos/antagonistas & inhibidores , Músculo Liso Vascular/efectos de los fármacos , Uncaria/química , Alcaloides/aislamiento & purificación , Animales , Aorta/citología , Aorta/efectos de los fármacos , Aorta/enzimología , Becaplermina , Células Cultivadas , ADN/biosíntesis , Relación Dosis-Respuesta a Droga , Medicamentos Herbarios Chinos/aislamiento & purificación , Alcaloides Indólicos , Músculo Liso Vascular/citología , Músculo Liso Vascular/enzimología , Fosforilación , Componentes Aéreos de las Plantas/química , Factor de Crecimiento Derivado de Plaquetas/farmacología , Proteínas Proto-Oncogénicas c-sis , RatasRESUMEN
DA-9102 isolated from Actinidia arguta is a candidate of natural medicine currently under Phase II clinical trial for atopic dermatitis in Korea. In this study, spontaneous dermatitis was induced by magnesium deficiency in hairless rats and this system was applied to assess the suppressive effects of DA-9102 on atopic dermatitis-like skin disease. Oral administration of DA-9102 at a dose of 100 mg/kg for 16 days substantially suppressed the occurrence of spontaneous dermatitis. Eczematous skin lesions, water loss and scratching behavior were significantly decreased by DA-9102 in a dose-dependent manner. Infiltration of inflammatory cells into the skin and pathologic remodeling of the epidermis and dermis were much less than the Mg-def. group. Results from flow cytometry analysis of peripheral blood mononuclear cells indicated that DA-9102 suppressed activation of leukocytes. The decrease in the number of CD45RA+ cells was accompanied by a lower level of IgE in DA-9102 treated rats, and the reduction in the number of CD11b+ cells by DA-9102 in both periphery and skin was significant. Further, DA-9102 not only suppressed the mRNA expression of T(H)2 cytokines including IL-4 and IL-10 in the lymph node but it also decreased the levels of inflammatory mediators such as nitric oxide and leukotriene B(4) (LTB(4)) in the serum. Taken together, these results suggest that DA-9102 is an orally applicable potent immune modulator capable of controlling the occurrence of atopic dermatitis-like skin disease.
Asunto(s)
Actinidia , Dermatitis Atópica/prevención & control , Medicamentos Herbarios Chinos/uso terapéutico , Fitoterapia , Animales , Antígeno CD11b/metabolismo , Dermatitis Atópica/etiología , Dermatitis Atópica/patología , Modelos Animales de Enfermedad , Medicamentos Herbarios Chinos/aislamiento & purificación , Inmunoglobulina E/sangre , Antígenos Comunes de Leucocito/metabolismo , Deficiencia de Magnesio/complicaciones , Masculino , Ratas , Ratas sin PeloRESUMEN
We investigated the effects of delta and theta brain wave frequency magnetic fields (3, 4, and 5) on mouse brain by detecting photonic oxidative stress makers; spontaneous photon emission (SPE) and lucigenin and tert-butyl hydroperoxide (TBHP) induced chemiluminescences (CL). For this purpose, Balb/C mice were exposed to 3, 4, and 5 Hz magnetic fields (MF) at 0.7 mT for 3 h, respectively. After that we monitored SPE and lucigenin and TBHP-induced CL of the homogenates of mice brains. There was a significant decrease in SPE in the 4 Hz MF-exposed group. Lucigenin-induced CL was also significantly decreased only in the 4 Hz MF-exposed group. TBHP-induced CL was also distinctively decreased by all frequencies, 3, 4, and 5 Hz MF exposures. These results showed that oxidative stress in a mouse brain was decreased by 4 Hz MF. We suggest that the application of 4 Hz MF will contribute to magnetic field therapy.
Asunto(s)
Encéfalo/metabolismo , Encéfalo/efectos de la radiación , Campos Electromagnéticos , Estrés Oxidativo/fisiología , Estrés Oxidativo/efectos de la radiación , Especies Reactivas de Oxígeno/metabolismo , Animales , Relación Dosis-Respuesta en la Radiación , Mediciones Luminiscentes , Masculino , Ratones , Ratones Endogámicos BALB C , Dosis de RadiaciónRESUMEN
Many investigators have encountered difficulty in clarifying the risks of exposure to bisphenol A (BPA), an endocrine disrupting chemical in epidemiological studies or animal experiments. In the present study, we developed biomarkers of BPA-induced proteomic alterations in immune organs of mouse offspring that were prenatally exposed to BPA (15 and 300 mg/L of drinking water; they were exposed to 8.9 +/- 1.8 mg of BPA/kg/day and 171.1 +/- 16.8 mg of BPA/kg/day, respectively) that were evaluated in terms of sex, age, and BPA-exposure levels. We performed 2D-gel analyses of samples from various tissues (thymus and spleen), exposure levels, sex, and ages (3- and 7-week-old) (N = 48), and found seven proteins that were altered in a BPA dose-dependent manner. Among them, we further studiedapo-AI, DPPIII, and VAT1, which are suspected to be associated with endocrine disorders. By performing Western blots, we confirmed BPA upregulation of all three proteins. Moreover, the apo-AI mRNA levels were increased in a BPA dose-dependent manner in 3- and 7-week-old female mice. Females and young offspring were somewhat more sensitive to protein alterations than others. Our study, which is based on proteome analyses, suggests that apo-AI, DPPIII, and VAT represent protein biomarkers for BPA and provide useful mechanistic clues for BPA-induced endocrine disruption.
Asunto(s)
Disruptores Endocrinos/toxicidad , Fenoles/toxicidad , Efectos Tardíos de la Exposición Prenatal , Biosíntesis de Proteínas/efectos de los fármacos , Proteómica , Bazo/efectos de los fármacos , Timo/efectos de los fármacos , Factores de Edad , Animales , Apolipoproteína A-I/biosíntesis , Compuestos de Bencidrilo , Biomarcadores/análisis , Western Blotting , Dipeptidil-Peptidasas y Tripeptidil-Peptidasas/biosíntesis , Relación Dosis-Respuesta a Droga , Regulación hacia Abajo , Electroforesis en Gel Bidimensional , Femenino , Ratones , Ratones Endogámicos ICR , Embarazo , Efectos Tardíos de la Exposición Prenatal/inducido químicamente , Efectos Tardíos de la Exposición Prenatal/inmunología , Biosíntesis de Proteínas/inmunología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores Sexuales , Bazo/crecimiento & desarrollo , Bazo/metabolismo , Timo/crecimiento & desarrollo , Timo/metabolismo , Regulación hacia Arriba , Proteínas de Transporte Vesicular de Monoaminas/biosíntesisRESUMEN
Mancozeb (MCZ) is known to have detrimental effects on the reproductive system, but the toxicity of MCZ on immune responses has not been systematically investigated. We investigated the effects of MCZ exposure on the activities of murine peritoneal macrophages through evaluation of MCZ-induced alteration of nitric oxide (NO) production and tumor necrosis factor-alpha (TNF-alpha) synthesis. Macrophages were examined ex vivo from mice orally treated with various doses of MCZ for 5 consecutive days per week for 4 weeks (subacute exposure, 250, 1000, 1500 mg/kg/day) followed by culture for 2 (TNF-alpha) or 3 days (NO) in the presence of LPS plus IFN-gamma. Macrophages from naive mice were also cultured with various concentrations of MCZ (0.05, 0.25, 0.5, 1 and 2 microg/mIL in the presence of LPS plus IFN-gamma for 2 (TNF-alpha) or 3 days (NO) in vitro. NO production was decreased with the in vitro exposure to all concentrations of MCZ. However, the amount of NO production by peritoneal macrophages from MCZ-subacutely exposed mice was increased in comparision with that of control group. In vitro, MCZ suppressed TNF-alpha secretion with significant reduction at 2 microg/mL MCZ. Conversely, TNF-alpha release was enhanced ex vivo. This study provides the substantial evidence on MCZ-induced alternation in macrophage activity. In order to clearly understand the contrasting effect of MCZ on peritoneal macrophage activity, it is necessary to further investigate the influence of major metabolite of MCZ (ETU) exposure on the NO production and TNF-alpha synthesis.
Asunto(s)
Macrófagos Peritoneales/efectos de los fármacos , Maneb/farmacología , Zineb/farmacología , Animales , Relación Dosis-Respuesta a Droga , Femenino , Macrófagos Peritoneales/metabolismo , Ratones , Ratones Endogámicos ICR , Nitritos/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Bisphenol A (BPA) is known to have detrimental effects on the reproductive system, but the toxicity of BPA on immune responses has not been systematically investigated. We investigated the effects of BPA exposure on the activities of murine peritoneal macrophages through evaluation of BPA-induced alteration of nitric oxide (NO) production, tumor necrosis factor-α (TNF-α) synthesis, and expression of co-stimulatory molecules B7. Macrophages were examined ex vivo from mice orally treated with various doses of BPA for 5 consecutive days per week for 4 weeks followed by culture for 2 or 4 days in the presence of lipopolysaccharides (LPS). Macrophages from naive mice were also stimulated with LPS ± BPA for 2 or 4 days. NO production was decreased with the in vitro exposure to 1, 10 and 100µM BPA. NO production was lower in the BPA-exposed mice than the control mice with all doses. In vitro, BPA suppressed TNF-α secretion with significant reduction at 10 and 100µM BPA. Similar findings were observed with the macrophages from the BPA-exposed mice. This study provides the substantial evidence on BPA-induced alteration in macrophage activity.
RESUMEN
The metabolism of bisphenol A (BPA), a suspected endocrine disruptor, should be considered for monitoring human exposure to BPA, because the conjugation with beta-D-glucuronide and sulfate reduces the estrogenic activity. In this study, BPA levels in 30 healthy Koreans (men, N=15, 42.6+/-2.4 years; women, N=15, 43.0+/-2.7 years) were analyzed from urine treated with/without beta-glucuronidase and/or sulfatase by an RP-HPLC with fluorescence detection. The total BPA concentrations including free BPA and the urinary conjugates were similar in men and women (2.82+/-0.73 and 2.76+/-0.54 ng ml(-1), respectively), but gender differences were found in the levels of urinary BPA conjugates. Men had significantly higher levels of BPA-glucuronide (2.34+/-0.85 ng ml(-1)) than women (1.00+/-0.34 ng ml(-1)), whereas women had higher levels of BPA-sulfate (1.20+/-0.32 ng ml(-1)) than men (0.49+/-0.27 ng ml(-1)).
Asunto(s)
Contaminantes Ambientales/orina , Glucuronidasa/química , Fenoles/química , Fenoles/orina , Sulfatasas/química , Urinálisis/métodos , Adulto , Compuestos de Bencidrilo , Cromatografía Líquida de Alta Presión/métodos , Femenino , Humanos , Masculino , Fenoles/metabolismo , Unión Proteica , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Espectrometría de Fluorescencia/métodosRESUMEN
Nonylphenol (NP) is widely used as a component of detergents, paints, pesticides, and many other formulated products. Several studies have demonstrated that NP is estrogenic in fish, avian, and mammalian cells. NP also competitively inhibits the binding of 17 beta-estradiol (E2) to the estrogen receptor (ER). However, there are relatively few in vivo data related to this issue in mammals. The aim of this study was to investigate the estrogenic activity of NP in animal models. We performed a 3-day uterotrophic assay using immature female rats for comparison with other endpoints of Tier I screening including vaginal opening (VO) in prepubertal intact female rats. For the uterotrophic assay, diethylstilbestrol (DES) (0.2 and 1.0 microg/kg) and p-NP (10, 25, 50, 100, and 200 mg/kg) were administered subcutaneously to immature Sprague-Dawley female rats for 3 consecutive days (postnatal days (PND) 20, 21, and 22). For the female pubertal onset assay, DES (0.2, 1.0, and 5.0 microg/kg) and p-NP (10, 50, and 100 mg/kg) were administered daily by oral gavage from 21 days of age for 20 days. In the uterotrophic assay, statistically significant increases in uterine wet weight were observed at doses of 100 and 200 mg/kg p-NP. DES (0.2 and 1.0 microg/kg) also significantly increased uterine weight compared to the vehicle control. In the female pubertal onset assay, the age of VO was advanced following oral exposure to DES (1.0 and 5.0 microg/kg) and p-NP (50 and 100 mg/kg). Estrous cyclicity was monitored in prepubertal rats from the day of VO to the day of necropsy. Irregular estrous cycles were observed in the groups treated with DES (5.0 microg/kg) and p-NP (50 and 100 mg/kg). High-dose DES (5.0 microg/kg) produced a persistent estrus state, whereas p-NP (50 and 100 mg/kg) increased the number of days in diestrus. Serum thyroxine (T(4)) concentrations were decreased in a dose-dependent manner by DES and p-NP treatment. A significant decrease in serum T(4) level was observed at high-dose DES (5.0 microg/kg) and p-NP (100 mg/kg). Serum TSH level was significantly increased by DES (5.0 microg/kg) treatment. Statistically significant decreases in ovarian weight were observed in female rats treated with DES (5.0 microg/kg) and p-NP (100 mg/kg). Our data demonstrate that p-NP can accelerate the onset of puberty and alter estrous cyclicity in prepubertal female rats at oral doses lower than the subcutaneous doses typically used in the uterotrophic assay. We therefore suggest that the female pubertal onset assay may be used as a sensitive testing method to detect environmental agents with weak estrogenic activity, but requires further research.