RESUMEN
Alzheimer's disease (AD) is an age-related neurodegenerative disorder characterized by cognitive deficits, which are accompanied by memory loss and cognitive disruption. Rhodiola sachalinensis (RSE) is a medicinal plant that has been used in northeastern Asia for various pharmacological activities. We attempted to carry out the bioconversion of RSE (Bio-RSE) using the mycelium of Bovista plumbe to obtain tyrosol-enriched Bio-RSE. The objective of this study was to investigate the effects of Bio-RSE on the activation of the cholinergic system and the inhibition of oxidative stress in mice with scopolamine (Sco)-induced memory impairment. Sco (1 mg/kg body weight, i.p.) impaired the mice's performance on the Y-maze test, passive avoidance test, and water maze test. However, the number of abnormal behaviors was reduced in the groups supplemented with Bio-RSE. Bio-RSE treatment improved working memory and avoidance times against electronic shock, increased step-through latency, and reduced the time to reach the escape zone in the water maze test. Bio-RSE dramatically improved the cholinergic system by decreasing acetylcholinesterase activity and regulated oxidative stress by increasing antioxidant enzymes (superoxide dismutase (SOD) and catalase (CAT)). The reduction in nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling in the brain tissue due to scopolamine was restored by the administration of Bio-RSE. Bio-RSE also significantly decreased amyloid-beta 1-42 (Aß1-42) and amyloid precursor protein (APP) expression. Moreover, the increased malondialdehyde (MDA) level and low total antioxidant capacity in Sco-treated mouse brains were reversed by Bio-RSE, and an increase in Nrf2 and HO-1 was also observed. In conclusion, Bio-RSE protected against Sco-induced cognitive impairment by activating Nrf2/HO-1 signaling and may be developed as a potential beneficial material for AD.
Asunto(s)
Enfermedad de Alzheimer , Rhodiola , Acetilcolinesterasa/metabolismo , Enfermedad de Alzheimer/inducido químicamente , Enfermedad de Alzheimer/tratamiento farmacológico , Enfermedad de Alzheimer/metabolismo , Animales , Antioxidantes/metabolismo , Colinérgicos/farmacología , Cognición , Aprendizaje por Laberinto , Trastornos de la Memoria/tratamiento farmacológico , Ratones , Micelio/metabolismo , Factor 2 Relacionado con NF-E2/metabolismo , Estrés Oxidativo , Alcohol Feniletílico/análogos & derivados , Extractos Vegetales/farmacología , Extractos Vegetales/uso terapéutico , Rhodiola/metabolismo , Escopolamina/farmacologíaRESUMEN
This study evaluated the use of an optical inspection system (OIS) to determine the freshness of mackerel (Scomber japonicus). The correlations between the light reflection intensity (LRI) of mackerel eyes (determined using an OIS) and the volatile basic nitrogen content (VBN) and K-value were analyzed. After unloading at the harbor, the mackerel were stored at 4 °C for 9 days and the VBN, K-value, and LRI were determined at 3-day intervals. During storage, the LRI, VBN, and K-value all increased. Furthermore, the LRI was correlated with the K-value and VBN. Therefore, although the LRI cannot be applied as an absolute standard for evaluating freshness, the LRI using an OIS is a suitable nondestructive method for evaluating freshness for quality and risk management in the processing industry when handling large numbers of fish.
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The effect of Rhodiola sachalinensis Boriss extract irradiated with 50 kGy gamma rays (HKC) on benign prostatic hyperplasia (BPH) was investigated. Seven-week-old male SD rats received a subcutaneous injection of 20 mg/kg of testosterone propionate (TP) to induce BPH. Then, the testosterone only group received testosterone, the testosterone + finasteride group received testosterone and finasteride (5 mg/kg), the testosterone + HKC group received testosterone and HKC extract (500 mg/kg). Prostate weight and the dihydrotestosterone (DHT) levels in serum or prostate tissue were determined. The mRNA expressions of 5-alpha reductase (AR) in prostate tissue were also measured. Compared to the control group, prostate weight was significantly improved in the TP group and decreased in the HKC and finasteride-treated groups. Furthermore, the mRNA expression of 5-AR in the prostate was significantly reduced in the HKC and finasteride-treated groups. Similarly, the expression levels of α-smooth muscle actin (α-SMA) and cytokeratin, which are associated with prostatic enlargement in the HKC and finasteride groups, were much lower than in the TP group. HKC treatment showed similar efficacy to finasteride treatment on rats with testosterone-induced BPH. HKC may be explored as a potential new drug for BPH treatment.
Asunto(s)
Colestenona 5 alfa-Reductasa/metabolismo , Rayos gamma , Extractos Vegetales/química , Extractos Vegetales/uso terapéutico , Hiperplasia Prostática/tratamiento farmacológico , Rhodiola/química , Testosterona/metabolismo , Testosterona/toxicidad , Animales , Colestenona 5 alfa-Reductasa/genética , Masculino , Hiperplasia Prostática/sangre , Ratas , Ratas Sprague-Dawley , Testosterona/sangreRESUMEN
Sortilin-related receptor 1 (SORL1) is a neuronal sorting protein that reduces amyloid precursor protein (APP) trafficking to secretases that generate amyloid beta (Aß). Although 6-shogaol, a constituent of ginger, has been reported to have anti-inflammatory and anti-oxidant effects on neuronal cells, research regarding the activation of SORL1 has not yet been reported. Here, we aimed to investigate whether 6-shogaol contributes to the increases in SORL1 that are related to Alzheimer's disease (AD). To clarify the effect of 6-shogaol as a possible activator of SORL1, we used SORL1 siRNA as a blockade of SORL1 in hippocampal neuronal cells (HT22). We found that SORL1 siRNA treatment naturally inhibited SORL1 and led to increases in ß-secretase APP cleaving enzyme (BACE), secreted APP-ß (sAPPß) and Aß. In contrast, 6-shogaol-mediated activation of SORL1 significantly downregulated BACE, sAPPß, and Aß in both in vitro HT22 cells and in vivo APPSw/PS1-dE9 Tg mice. Therefore, SORL1 activation by 6-shogaol provides neuronal cell survival through the inhibition of Aß production. These results indicate that 6-shogaol should be regarded as an SORL1 activator and a potential preventive agent for the treatment of AD.
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Precursor de Proteína beta-Amiloide/metabolismo , Proteínas Amiloidogénicas/metabolismo , Catecoles/administración & dosificación , Proteínas de Transporte de Membrana/metabolismo , Neuronas/metabolismo , Receptores de LDL/metabolismo , Transducción de Señal/fisiología , Animales , Línea Celular , Relación Dosis-Respuesta a Droga , Humanos , Ratones , Mutágenos/administración & dosificación , Neuronas/efectos de los fármacos , Transducción de Señal/efectos de los fármacosRESUMEN
This study was conducted to improve sensory quality of Jokbal (Korean Pettitoes) made from frozen pig feet by addition of herbal mixture (glasswort, raspberry and Sansa powders). After adding herbal mixture, lipid oxidation (2-thiobarbituric acid values, TBARS), sensory property, and textural property were determined. Herbs were individually added into cooking soup at concentration of 6% (low concentration treatment, LCT) or 12% (high concentration treatment, HCT) of raw pig feet. Refrigerated pig feet were used as control. Thawed feet without any herbal mixture were used as freezing treatment (FT). TBARS in LCT or HCT were lower than that in FT, and showed the similar to that in Control. Addition of the herbal mixture was effective in improving the flavor and textural property of thawed feet by inhibiting lipid oxidation and protein denaturation in a dose-dependent manner.
RESUMEN
6-Shogaol can be extracted from ginger and has been shown to exert anti-inflammatory and antioxidant activities, which are potentially relevant to the treatment of central nervous system disorders. Oxidative stress and inflammation are closely associated with ischemic injury and can eventually result in neuronal death. The aim of this study was to evaluate if 6-shogaol exerts neuroprotective activity. To this end, we determined its effects on oxidative stress and inflammation in a mouse model of middle cerebral artery occlusion (MCAO)-induced brain damage. In this model, MCAO was induced in C57BL/6 mice (30-35g, 9 weeks) for 1h, followed by 24h reperfusion. Mice were treated orally with 6-shogaol (0.1ml, 5 or 20mg/kg) once daily for 7 consecutive days prior to MCAO. We found that 6-shogaol significantly reduced neurological deficit scores and the mean infarct area. Moreover, 6-shogaol improved the behavioral deficits in the MCAO group. In addition, 6-shogaol pretreatment dampened MCAO-mediated production of reactive oxygen species and inflammatory cytokines. Mechanistic studies revealed that 6-shogaol inhibits the cysteinyl leukotriene 1 receptor (CysLT1R) and mitogen-activated protein kinase (MAPK) signaling proteins, thus providing a potential pharmacological mechanism for our observations. These results suggest that 6-shogaol can ameliorate the outcomes of MCAO and could thus be used as a potential preventive of stroke.
Asunto(s)
Catecoles/farmacología , Infarto de la Arteria Cerebral Media/tratamiento farmacológico , Infarto de la Arteria Cerebral Media/metabolismo , Estrés Oxidativo/efectos de los fármacos , Animales , Conducta Animal/efectos de los fármacos , Catecoles/uso terapéutico , Citocinas/biosíntesis , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Infarto de la Arteria Cerebral Media/complicaciones , Infarto de la Arteria Cerebral Media/patología , Inflamación/complicaciones , Masculino , Malondialdehído/metabolismo , Ratones , Ratones Endogámicos C57BL , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Receptores de Leucotrienos/metabolismo , Transducción de Señal/efectos de los fármacosRESUMEN
Although 6-shogaol, a constituent of ginger, has been reported to have anti-inflammatory and anti-oxidant effects on neuronal cells, the effects of 6-shogaol on Alzheimer's disease (AD) have not yet been investigated. Here we aimed to determine whether 6-shogaol exerts neuroprotective effects against AD. Specifically, we investigated the effects of 6-shogaol on the cysteinyl leukotriene 1 receptor (CysLT1R), a major factor in AD pathogenesis. Moreover, we clarified the relationship between CysLT1R and cathepsin B, a cysteine protease. We used in vitro and in vivo models to determine whether 6-shogaol inhibits CysLT1R/cathepsin B in an amyloid-beta (Aß; 1-42)-induced model of neurotoxicity. We first confirmed that CysLT1R and cathepsin B are upregulated by Aß (1-42) and that CysLT1R activation induces cathepsin B. In contrast, we found that 6-shogaol-mediated inhibition of CysLT1R downregulates cathepsin B in both in vitro and in vivo models. Furthermore, we found that 6-shogaol-mediated inhibition of CysLT1R/cathepsin B reduces Aß deposition in the brain and ameliorates behavioral deficits in APPSw/PS1-dE9 Tg mice. Our results indicate that 6-shogaol is a CysLT1R/cathepsin B inhibitor and is a novel potential therapeutic agent for the treatment of various neurodegenerative diseases, including AD.
Asunto(s)
Enfermedad de Alzheimer/prevención & control , Péptidos beta-Amiloides/antagonistas & inhibidores , Catecoles/farmacología , Catepsina B/antagonistas & inhibidores , Fragmentos de Péptidos/antagonistas & inhibidores , Receptores de Leucotrienos/fisiología , Animales , Reacción de Prevención/efectos de los fármacos , Encéfalo/efectos de los fármacos , Encéfalo/enzimología , Encéfalo/metabolismo , Línea Celular , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Ratones , Ratones Endogámicos C57BL , Ratones TransgénicosRESUMEN
This study was conducted to investigate the effects of kimchi powder and onion peel extract on the quality characteristics of emulsion sausage manufactured with irradiated pork. The emulsion sausages were formulated with 2% kimchi powder and/or 0.05% onion peel extract. The changes in pH value of all treatments were similar, depending on storage periods. The addition of kimchi powder increased the redness and yellowness of the emulsion sausage. The addition of onion peel extract decreased the thiobarbituric acid reactive substances value of the emulsion sausages prepared with irradiated pork. The volatile basic nitrogen value of the emulsion sausage prepared with kimchi powder was the highest, whereas that of the emulsion sausage prepared with onion peel extract was the lowest. The treatment without kimchi powder or onion peel extract and the treatments prepared with onion peel extract showed lower microbial populations than the other treatment. Sensory evaluations indicated that a higher acceptability was attained when kimchi powder was added to the emulsion sausages manufactured with irradiated pork. In conclusion, our results suggest that combined use of kimchi powder and onion peel extract could improve quality characteristics and shelf stability of the emulsion sausage formulated with irradiated pork during chilled storage.
RESUMEN
The purpose of this study was to investigate the effect of lactic acid bacteria (LAB) cell wall extract on the proliferation and cytokine production of immune cells to select suitable probiotics for space food. Ten strains of LAB (Lactobacillus bulgaricus, L. paracasei, L. casei, L. acidophilus, L. plantarum, L. delbruekii, Lactococcus lactis, Streptococcus thermophilus, Bifidobacterium breve, and Pedicoccus pentosaceus) were sub-cultured and further cultured for 3 d to reach 7-10 Log colony-forming units (CFU)/mL prior to cell wall extractions. All LAB cell wall extracts failed to inhibit the proliferation of BALB/c mouse splenocytes or mesenteric lymphocytes. Most LAB cell wall extracts except those of L. plantarum and L. delbrueckii induced the proliferation of both immune cells at tested concentrations. In addition, the production of TH1 cytokine (IFN-γ) rather than that of TH2 cytokine (IL-4) was enhanced by LAB cell wall extracts. Of ten LAB extracts, four (from L. acidophilus, L. bulgaricus, L. casei, and S. thermophiles) promoted both cell proliferating and TH1 cytokine production. These results suggested that these LAB could be used as probiotics to maintain immunity and homeostasis for astronauts in extreme space environment and for general people in normal life.
RESUMEN
This study evaluated the effectiveness of irradiated porcine tendon xenografts for lateral collateral ligament (LCL) reconstruction. Twenty healthy adult beagle dogs underwent LCL reconstruction using irradiated porcine tendons treated with poly-gamma-glutamic acid. Serological and histological assessments were performed to evaluate host immunological response at 3 and 12 months after surgery. The healing and functional integrity of the LCL reconstructions were assessed by mechanical testing and gait analysis. Histological assessment of the porcine xenografts showed gradual host cellular infiltration and graft collagen remodeling during the healing process. Porcine xenografts showed angiogenesis and no signs of inflammatory reaction. Additionally, biomechanical and gait evaluations supported graft functional integration with no differences between normal and porcine xenograft reconstruction at 12 months after surgery. Irradiated porcine xenografts showed greater cellular responses and healing properties in short- and long-term evaluations. Irradiated porcine tendons appear to be useful as xenografts for the reconstruction of damaged ligaments.
Asunto(s)
Ligamentos Colaterales/cirugía , Marcha/fisiología , Xenoinjertos/fisiología , Procedimientos de Cirugía Plástica/veterinaria , Tendones/trasplante , Trasplante Heterólogo/veterinaria , Animales , Fenómenos Biomecánicos , Perros , Rayos gamma , Xenoinjertos/anatomía & histología , Xenoinjertos/efectos de la radiación , Porcinos , Tendones/efectos de la radiación , Cicatrización de Heridas/fisiología , Cicatrización de Heridas/efectos de la radiaciónRESUMEN
The present study investigated the effects of viscothionin, a compound isolated from Korean mistletoe (Viscum album coloratum), on nonalcoholic fatty liver disease (NAFLD) in both in vitro and in vivo models. A connection was discovered between viscothionin and the adenosine monophosphate-activated protein kinase (AMPK) signaling pathway, which is involved in lipid metabolism. Viscothionin was shown to significantly attenuate lipid accumulation in HepG2 cells treated with oleic acid, which induces lipid accumulation. Moreover, the phosphorylation of AMPK and acetyl-coenzyme A carboxylase in HepG2 cells was increased by viscothionin treatment. Viscothionin was orally administered to high fat diet-induced obese mice and subsequently histopathological analysis associated with AMPK signaling pathways was evaluated. A significant reduction in the extent of hepatic steatosis was revealed in viscothionin-treated obese mice. Thus, viscothionin mediates its beneficial effects on NAFLD via AMPK signaling pathways, suggesting that it may be a potential target for novel NAFLD treatments.
Asunto(s)
Proteínas Quinasas Activadas por AMP/metabolismo , Muérdago/química , Enfermedad del Hígado Graso no Alcohólico/tratamiento farmacológico , Péptidos/administración & dosificación , Extractos Vegetales/administración & dosificación , Proteínas Quinasas Activadas por AMP/genética , Acetil-CoA Carboxilasa/genética , Acetil-CoA Carboxilasa/metabolismo , Animales , Activadores de Enzimas , Acido Graso Sintasa Tipo I/genética , Acido Graso Sintasa Tipo I/metabolismo , Células Hep G2 , Humanos , Metabolismo de los Lípidos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Obesos , Enfermedad del Hígado Graso no Alcohólico/enzimología , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Fosforilación , República de CoreaRESUMEN
The aim of this study was to clarify the efficacy of procyanidin C1 (Pro C1) for modulating vascular tone. Pro C1 induced a potent vasorelaxant effect on phenylephrine-constricted endothelium-intact thoracic aortic rings, but had no effect on denuded thoracic aortic rings. Moreover, Pro C1 caused a significant increase in nitric oxide (NO) production in endothelial cells. Pro C1-induced vasorelaxation and Pro C1-induced NO production were significantly decreased in the presence of a nonspecific potassium channel blocker (tetraethylammonium chloride [TEA]), an endothelial NO synthase inhibitor (N(G)-monomethyl-L-arginine [L-NMMA]), and a store-operated calcium entry inhibitor (2-aminoethyl diphenylborinate [2-APB]). Pro C1-induced vasorelaxation was also completely abolished by an inhibitor of soluble guanyl cyclase, which suggests that the Pro C1 effects observed involved cyclic guanosine monophosphate (cGMP) production. Interestingly, Pro C1 significantly enhanced basal cGMP levels. Taken together, these results indicate that Pro C1-induced vasorelaxation is associated with the activation of the calcium-dependent NO/cGMP pathway, involving potassium channel activation. Thus, Pro C1 may represent a novel and potentially therapeutically relevant compound for the treatment of cardiovascular diseases.
Asunto(s)
Biflavonoides/farmacología , Catequina/farmacología , GMP Cíclico/metabolismo , Óxido Nítrico/metabolismo , Proantocianidinas/farmacología , Transducción de Señal/efectos de los fármacos , Vasodilatación/efectos de los fármacos , Animales , Aorta Torácica/efectos de los fármacos , Compuestos de Boro/farmacología , Células Cultivadas , Células Endoteliales/efectos de los fármacos , Guanilato Ciclasa/antagonistas & inhibidores , Guanilato Ciclasa/metabolismo , Técnicas In Vitro , Masculino , Óxido Nítrico Sintasa de Tipo III/metabolismo , Bloqueadores de los Canales de Potasio/farmacología , Ratas , Ratas Sprague-Dawley , Tetraetilamonio/farmacología , Vasodilatadores/farmacología , omega-N-Metilarginina/farmacologíaRESUMEN
The principal objective of this study was to assess the DNA damage in a normal cell line system after exposure to 60 Hz of extremely low frequency magnetic field (ELF-MF) and particularly in combination with various external factors, via comet assays. NIH3T3 mouse fibroblast cells, WI-38 human lung fibroblast cells, L132 human lung epithelial cells, and MCF10A human mammary gland epithelial cells were exposed for 4 or 16 h to a 60-Hz, 1 mT uniform magnetic field in the presence or absence of ionizing radiation (IR, 1 Gy), H(2)O(2) (50 µM), or c-Myc oncogenic activation. The results obtained showed no significant differences between the cells exposed to ELF-MF alone and the unexposed cells. Moreover, no synergistic or additive effects were observed after 4 or 16 h of pre-exposure to 1 mT ELF-MF or simultaneous exposure to ELF-MF combined with IR, H(2)O(2), or c-Myc activation.
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Daño del ADN , Campos Electromagnéticos/efectos adversos , Peróxido de Hidrógeno/farmacología , Proteínas Proto-Oncogénicas c-myc/genética , Animales , Línea Celular , Relación Dosis-Respuesta a Droga , Relación Dosis-Respuesta en la Radiación , Expresión Génica , Humanos , Ratones , Oncogenes/genética , Radiación IonizanteRESUMEN
Ground lean pork was irradiated by an electron beam or X-rays to compare the effects of two types of radiation generated by a linear accelerator on the quality of Bologna sausage as a model meat product. Raw ground lean pork was vacuum packaged at a thickness of 1.5 cm and irradiated at doses of 2, 4, 6, 8, or 10 kGy by an electron beam (2.5 MeV) or X-rays (5 MeV). Solubility of myofibrillar proteins, bacterial counts, and thiobarbituric acid reactive substance (TBARS) values were determined for raw meat samples. Bologna sausage was manufactured using the irradiated lean pork, and total bacterial counts, TBARS values, and quality properties (color differences, cooking yield, texture, and palatability) were determined. Irradiation increased the solubility of myofibrillar proteins in a dose-dependent manner (p<0.05). Bacterial contamination of the raw meat was reduced as the absorbed dose increased, and the reduction was the same for both radiation types. Differences were observed only between irradiated and non-irradiated samples (p<0.05). X-ray irradiation may serve as an alternative to gamma irradiation and electron beam irradiation.
RESUMEN
This study was conducted to compare the effects of two forms of radiation (electron and X-ray; generated by an electron beam accelerator) on the conformation and antigenic properties of hen's egg albumin, ovalbumin (OVA), which was used as a model protein. OVA solutions (2.0 mg/mL) were individually irradiated by electron beam or X-ray at the absorbed doses of 0 (control), 2, 4, 6, 8, and 10 kGy. No differences between the two forms of radiation on the structural properties of OVA were shown by spectrometric and electrophoretic analyses. The turbidity of OVA solution increased and the main OVA bands on polyacrylamide gels disappeared after irradiation, regardless of the radiation source. In competitive indirect enzyme-linked immunosorbent assay, OVA samples irradiated by electron beam or X-ray showed different immunological responses in reactions with monoclonal and polyclonal antibodies (immunoglobulin G) produced against non-irradiated OVA. The results indicate that electron beam irradiation and X-ray irradiation produced different patterns of structural changes to the OVA molecule.
RESUMEN
The combined effects of kimchi powder (KP) and smoking (SM) on the quality characteristics and shelf life of cooked sausage prepared with 10 kGy irradiated pork (IP) were studied. The cooked sausages were formulated with single or combined treatment of 0.5% KP and SM (70°C for 30 min). IP+KP+SM treatment showed increased redness, hardness, gumminess, and chewiness, but decreased cooking yield. As for sensory evaluation, treatment with both KP and SM was effective to mask the radiolytic off-flavor. Moreover, the cooked sausage treated with both KP and SM had the highest color, flavor, and overall acceptance (P<0.05), where the differences of flavor patterns were verified by using an electronic nose. During chilled storage for 4 weeks, the combined treatment is effective to retard lipid oxidation, formation of volatile compound, and total microbial number due to the addition of KP. Therefore, usages of KP and SM can provide improved quality characteristics and shelf life of cooked sausage prepared with IP.
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Irradiación de Alimentos/métodos , Conservantes de Alimentos/análisis , Calidad de los Alimentos , Productos de la Carne/análisis , Productos de la Carne/efectos de la radiación , Animales , Fenómenos Químicos , Frío , Color , Culinaria , Embalaje de Alimentos , Conservación de Alimentos , Almacenamiento de Alimentos , Humanos , Concentración de Iones de Hidrógeno , Odorantes , Porcinos , Gusto , Sustancias Reactivas al Ácido Tiobarbitúrico/análisisRESUMEN
Polyphenolic compounds have been found to possess a wide range of physiological activities that may contribute to their beneficial effects against inflammation-related diseases; however, the molecular mechanisms underlying this anti-inflammatory activity are not completely characterized, and many features remain to be elucidated. In this study, we investigated the molecular basis for the down-regulation of toll-like receptor 4 (TLR4) signal transduction by procyanidin dimer B2 (Pro B2) in macrophages. Pro B2 markedly elevated the expression of the interleukin (IL)-1 receptor-associated kinase (IRAK)-M protein, a negative regulator of TLR signaling. Lipopolysaccharide (LPS)-induced expression of cell surface molecules (CD80, CD86, and MHC class I/II) and production of pro-inflammatory cytokines (tumor necrosis factor-α, IL-1ß, IL-6, and IL-12p70) were inhibited by Pro B2, and this action was prevented by IRAK-M silencing. In addition, Pro B2-treated macrophages inhibited LPS-induced activation of mitogen-activated protein kinases such as extracellular signal-regulated kinase 1/2, p38, and c-Jun N-terminal kinase and the translocation of nuclear factor κB and p65 through IRAK-M. We also found that Pro B2-treated macrophages inactivated naïve T cells by inhibiting LPS-induced interferon-γ and IL-2 secretion through IRAK-M. These novel findings provide new insights into the understanding of negative regulatory mechanisms of the TLR4 signaling pathway and the immune-pharmacological role of Pro B2 in the immune response against the development and progression of many chronic diseases.
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Biflavonoides/farmacología , Catequina/farmacología , Quinasas Asociadas a Receptores de Interleucina-1/biosíntesis , Macrófagos/metabolismo , Proantocianidinas/farmacología , Receptor Toll-Like 4/metabolismo , Animales , Línea Celular , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/fisiología , Macrófagos/efectos de los fármacos , RatonesRESUMEN
This study evaluated the effect of gamma irradiation on the reduction of the toxicity of mistletoe lectin using both in vitro and in vivo models. To extract the lectin from mistletoe, an (NH4)2SO4 precipitation method was employed and the precipitant purified using a Sepharose 4B column to obtain the pure lectin fraction. Purified lectin was then gamma-irradiated at doses of 0, 5, 10, 15, and 20 kGy, or heated at 100 °C for 30 min. Toxic effects of non-irradiated, irradiated, and heat-treated lectins were tested using hemagglutination assays, cytotoxicity assays, hepatotoxicity, and a mouse survival test and immunological response was tested using cytokine production activity. Hemagglutination of lectin was remarkably decreased (P < 0.05) by irradiation at doses exceeding 10 kGy and with heat treatment. However, lectin irradiated with 5 kGy maintained its hemagglutination activity. The cytotoxicity of lectin was decreased by irradiation at doses over 5 kGy and with heat treatment. In experiments using mouse model, glutamate oxaloacetate transaminase (GOT) and glutamic pyruvic transaminase (GPT) levels were decreased in the group treated with the 5 kGy irradiated and heat-treated lectins as compared to the intact lectin, and it was also shown that 5 kGy irradiated and heat-treated lectins did not cause damage in liver tissue or mortality. In the result of immunological response, tumor necrosis factor (TNF-α) and interleukin (IL-6) levels were significantly (P < 0.05) increased in the 5 kGy gamma-irradiated lectin treated group. These results indicate that 5 kGy irradiated lectin still maintained the immunological response with reduction of toxicity. Therefore, gamma-irradiation may be an effective method for reducing the toxicity of lectin maintaining the immune response.
RESUMEN
Numerous studies have shown various relationships between foods with a high nutritional value and a robust immune response, particularly studies that have focused on host protection and cytokine networks. This study aimed to clarify the role played by the procyanidin trimer C1 in innate and adaptive immunity. Procyanidin C1 did not exert cytotoxicity at concentrations ranging from 7.8 to 62.5 µg/ml in macrophage cells; therefore, concentration of 62.5 µg/ml was used as the maximum dose of procyanidin C1 throughout subsequent experiments. Procyanidin C1 enhanced inducible nitric oxide synthase-mediated nitric oxide production in a concentration-dependent manner. In addition, procyanidin C1 functionally induced macrophage activation by augmenting the expression of cell surface molecules (CD80, CD86, and MHC II) and proinflammatory cytokine production (tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, and IL-6) via activation of mitogen-activated protein kinase (MAPK), e.g., p38, ERK, and JNK and nuclear factor (NF)-κB signaling pathways. Interestingly, procyanidin C1 effectively polarized T helper type 1 (Th1) by secreting Th1-mediated cytokines (interferon-γ, IL-12p70, and IL-2) and inducing splenocyte proliferation, indicating that procyanidin C1 contributes to Th1 polarization of the immune response. Accordingly, these findings confirms that the procyanidin C1 induces macrophage activation via NF-κB and MAPK pathways, leading to Th1 polarization in murine splenocytes, which suggests that procyanidin C1 regulates innate and adaptive immunity by macrophage activation and Th1 polarization.
Asunto(s)
Biflavonoides/química , Biflavonoides/farmacología , Catequina/química , Catequina/farmacología , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Activación de Macrófagos/efectos de los fármacos , FN-kappa B/metabolismo , Polimerizacion , Proantocianidinas/química , Proantocianidinas/farmacología , Bazo/citología , Animales , Células de la Médula Ósea/citología , Línea Celular , Proliferación Celular/efectos de los fármacos , Citocinas/biosíntesis , Macrófagos/citología , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Macrófagos/metabolismo , Ratones , Óxido Nítrico/biosíntesis , Bazo/inmunología , Células TH1/citología , Células TH1/efectos de los fármacosRESUMEN
Natural products and dietary components rich in polyphenols have been shown to reduce inflammation; however, the molecular mechanisms underlying this anti-inflammatory activity are not completely characterized, and many features remain to be elucidated. This research was carried out to clarify the potential role of procyanidin trimer C1 in the anti-inflammatory effect of polyphenols. Procyanidin C1 inhibited inducible nitric oxide synthase-mediated nitric oxide production and the release of pro-inflammatory cytokines (interleukin-6 and tumor necrosis factor-α) in lipopolysaccharide (LPS)-induced macrophages. Treatment with procyanidin C1 resulted in a significant decrease in prostaglandin E2 and cyclooxygenase-2 levels, as well as the expression of cell surface molecules (CD80, CD86, and MHC class II), which was induced by LPS. Furthermore, our data demonstrated that the anti-inflammatory effect of procyanidin C1 occurs through inhibition of mitogen-activated protein kinase (p38 and c-Jun N-terminal kinase) and nuclear factor-κB signaling pathways. These 2 factors play a major role in controlling inflammation, through toll-like receptor 4, suggesting that procyanidin C1 plays a potent role in promoting anti-inflammatory activity in macrophages. These results represent a novel and effective therapeutic intervention for the treatment of inflammatory disease.