RESUMEN
1,5-Anhydro-D-fructose (1,5-AF) is a bioactive monosaccharide that is produced by the glycogenolysis in mammalians and is metabolized to 1,5-anhydro-D-glucitol (1,5-AG). 1,5-AG is used as a marker of glycemic control in diabetes patients. 1,5-AF has a variety of physiological activities, but its effects on energy metabolism, including feeding behavior, are unclarified. The present study examined whether 1,5-AF possesses the effect of satiety. Peroral administration of 1,5-AF, and not of 1,5-AG, suppressed daily food intake. Intracerebroventricular (ICV) administration of 1,5-AF also suppressed feeding. To investigate the neurons targeted by 1,5-AF, we investigated c-Fos expression in the hypothalamus and brain stem. ICV injection of 1,5-AF significantly increased c-Fos positive oxytocin neurons and mRNA expression of oxytocin in the paraventricular nucleus (PVN). Moreover, 1,5-AF increased cytosolic Ca2+ concentration of oxytocin neurons in the PVN. Furthermore, the satiety effect of 1,5-AF was abolished in oxytocin knockout mice. These findings reveal that 1,5-AF activates PVN oxytocin neurons to suppress feeding, indicating its potential as the energy storage monitoring messenger to the hypothalamus for integrative regulation of energy metabolism.
Asunto(s)
Oxitocina , Núcleo Hipotalámico Paraventricular , Ratones , Animales , Núcleo Hipotalámico Paraventricular/metabolismo , Oxitocina/metabolismo , Hipotálamo/metabolismo , Neuronas/metabolismo , Mamíferos/metabolismoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Ninjin'yoeito (NYT), a traditional Japanese Kampo medicine consisting of 12 herbs, has been reported to improve cognitive dysfunction, depression, and neurological recovery in patients with neurovascular diseases such as Alzheimer's disease and stroke. Several studies have reported that the NYT components exert neurotrophic, neurogenic, and neuroprotective effects. In addition, exercise enhances neuroprotection and functional recovery after stroke. Rehabilitative exercises and pharmacological agents induce neurophysiological plasticity, leading to functional recovery in stroke patients. These reports indicate that NYT treatment and exercise may promote functional recovery following stroke through their beneficial effects. However, no study has determined the effects of NYT and the possible mechanisms of neurorepair and functional recovery after stroke. AIM OF THE STUDY: This study aimed to investigate the combined effects of NYT and exercise on neuroprotection and functional recovery and the underlying mechanisms in a rat ischemic stroke model. MATERIALS AND METHODS: Stroke was induced with 60-min middle cerebral artery occlusion (MCAO) followed by reperfusion in adult male Sprague-Dawley rats. After stroke, the rats were assigned to four groups: ischemia reperfusion (IR), NYT, exercise (Ex), and NYT + Ex. NYT-treated rats were fed a diet containing 1% NYT one day after stroke. Exercise was performed using a motorized treadmill for 5 days a week (8-15 m/min, 20 min/day), starting 3 days after stroke. The NYT treatment and exercise were continued for 4 weeks after the stroke. Infarct volume, neurological deficits, sensorimotor functions, expression of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), tropomyosin receptor kinase A (TrkA) and B (TrkB), caspase-3 activity, and the p-Akt/Akt ratio were examined by immunohistochemistry and western blotting. RESULTS: Compared to the IR group, all treated groups indicated reduced infarct volumes. The NYT + Ex group showed significantly improved waking time and beam walking score compared with the IR group. The expression of NGF/TrkA/p-TrkA and BDNF/TrkB was significantly increased in the NYT + Ex group compared with those in the IR group, whereas the number of caspase-3 positive cells around the lesion was significantly lower in the NYT + Ex group than in the IR group. In addition, the ratio of p-Akt/Akt was significantly higher in the NYT + Ex group than in the IR group. CONCLUSIONS: This study suggests that NYT in combination with exercise provides neuroprotective effects and improves sensorimotor function by stimulating NGF/TrkA and BDNF/TrkB, and by activating the Akt pathway in ischemic stroke of rats. NYT may be an effective adjunctive agent in post-stroke rehabilitation.
Asunto(s)
Accidente Cerebrovascular Isquémico , Medicina Kampo , Fármacos Neuroprotectores , Condicionamiento Físico Animal , Animales , Masculino , Ratas , Factor Neurotrófico Derivado del Encéfalo , Caspasa 3 , Infarto , Accidente Cerebrovascular Isquémico/tratamiento farmacológico , Factor de Crecimiento Nervioso , Fármacos Neuroprotectores/farmacología , Fármacos Neuroprotectores/uso terapéutico , Proteínas Proto-Oncogénicas c-akt , Ratas Sprague-DawleyRESUMEN
Physical frailty is an aging-related clinical syndrome involving decreases in body weight, mobility, activity, and walking speed that occurs in individuals with sarcopenia and is accelerated by increased oxidative stress. Ninjin'yoeito, a traditional Japanese Kampo medicine, is used for treating conditions, including anemia and physical weakness. Here, we investigated whether ninjin'yoeito could improve physical frailty by controlling oxidative stress in the senescence-accelerated mouse prone 8 (SAMP8) model. First, SAMP8 mice were divided into two groups, ninjin'yoeito treated and untreated, with the former consuming a diet containing 3% ninjin'yoeito from 3 months of age. At 7 months of age, body weight, motor function, locomotor activity, and mean walking speed were measured. Subsequently, mice were euthanized and measured for muscle weight, 8-hydroxy-2'-deoxyguanosine levels in muscle and brain, and cleaved caspase-3 expression in brain. The results showed reductions in weight, locomotor function, locomotion, and average walking speed in the untreated group, which were significantly improved by ninjin'yoeito. Furthermore, 8-hydroxy-2'-deoxyguanosine levels were reduced in muscle and brain from ninjin'yoeito-treated mice, compared with the levels in untreated mice; cleaved caspase-3 expression was similarly reduced in brain from the treated mice, indicating reduced apoptosis. Our findings suggest that ninjin'yoeito inhibits sarcopenia-based physical frailty through its antioxidant effects.
Asunto(s)
Fragilidad , Sarcopenia , 8-Hidroxi-2'-Desoxicoguanosina , Animales , Antioxidantes , Peso Corporal , Caspasa 3 , Modelos Animales de Enfermedad , Medicamentos Herbarios Chinos , Ratones , Sarcopenia/tratamiento farmacológicoRESUMEN
The gut microbiota has tremendous potential to affect the host's health, in part by synthesizing vitamins and generating nutrients from food that is otherwise indigestible by the host. 1,5-Anhydro-D-fructose (1,5-AF) is a monosaccharide with a wide range of bioactive potentials, including anti-oxidant, anti-inflammatory, and anti-microbial effects. Based on its potential benefits and minimal toxicity, it is anticipated that 1,5-AF will be used as a dietary supplement to support general health. However, the effects of 1,5-AF on the gut microbiota are yet to be clarified. Here, using an unbiased metagenomic approach, we profiled the bacterial taxa and functional genes in the caecal microbiota of mice fed a diet containing either 2% 1,5-AF or a reference sweetener. Supplementation with 1,5-AF altered the composition of the gut microbiota, enriching the proportion of Faecalibacterium prausnitzii. 1,5-AF also altered the metabolomic profile of the gut microbiota, enriching genes associated with nicotinamide adenine dinucleotide biosynthesis. These findings support the potential benefits of 1,5-AF, but further studies are required to clarify the impact of 1,5-AF on health and disease.
Asunto(s)
Fructosa/análogos & derivados , Microbioma Gastrointestinal , Animales , Dieta , Suplementos Dietéticos , Fructosa/metabolismo , Fructosa/farmacología , Microbioma Gastrointestinal/efectos de los fármacos , Metagenoma , Metagenómica/métodos , Ratones , NAD/biosíntesis , Nutrientes/biosíntesis , Vitaminas/biosíntesisRESUMEN
PURPOSE: To determine the effect of histones on corneal endothelial cells generated during cataract surgery. SETTING: Kagoshima University Hospital, Kagoshima, Japan. DESIGN: Experimental study. METHODS: Standard phacoemulsification was performed on enucleated pig eyes. Histones in the anterior segment of the eye were determined by immunohistochemistry. Cultured human corneal endothelial cells were exposed to histones for 18 hours, and cell viability was determined by 2-(2-methoxy-4-nitrophenyl)-3-(4-nitro-phenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt assay. The concentration of interleukin-6 (IL-6) in the culture medium of human corneal endothelial cells was measured using enzyme-linked immunosorbent assay. The effects of signal inhibitors U0126, SB203580, and SP600125 were evaluated. The protective effect of hyaluronan against histones was evaluated in human corneal endothelial cells with and without hyaluronan. RESULTS: Cellular debris containing histones was observed in the anterior chamber of pig eyes after phacoemulsification. Exposure of human corneal endothelial cells to 50 µg/mL of histones or more led to cytotoxic effects. The IL-6 concentration was significantly increased dose dependently after exposure of human corneal endothelial cells to histones (P<.01). The histone-induced IL-6 production was significantly decreased by extracellular signal-regulated kinases 1/2 and p-38 mitogen-activated protein kinase inhibitors (P<.01). Co-incubation of hyaluronan and histones caused formation of histone aggregates, decreased the cytotoxic effects of the histones, and blocked the increase in IL-6 (P<.01). CONCLUSIONS: Histones were released extracellularly during phacoemulsification and exposure of human corneal endothelial cells to histones increased the IL-6 secretion. The intraoperative use of hyaluronan may decrease the cytotoxic effects of histones released during cataract surgery. FINANCIAL DISCLOSURE: No author has a financial or proprietary interest in any material or method mentioned.
Asunto(s)
Endotelio Corneal/efectos de los fármacos , Histonas/toxicidad , Ácido Hialurónico/farmacología , Facoemulsificación , Viscosuplementos/farmacología , Animales , Cámara Anterior/metabolismo , Supervivencia Celular , Células Cultivadas , Citoprotección/efectos de los fármacos , Endotelio Corneal/metabolismo , Inhibidores Enzimáticos/farmacología , Ensayo de Inmunoadsorción Enzimática , Técnica del Anticuerpo Fluorescente Indirecta , Histonas/metabolismo , Humanos , Interleucina-6/metabolismo , Proteínas Quinasas JNK Activadas por Mitógenos/antagonistas & inhibidores , Proteínas Quinasas Activadas por Mitógenos/antagonistas & inhibidores , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Porcinos , Proteína de la Zonula Occludens-1/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/antagonistas & inhibidoresRESUMEN
OBJECTIVES: The usefulness of omega-3 lipid emulsions has been extensively studied. The objectives of the present study were to examine the effect of an omega-3 lipid emulsion in reducing oxidative stress in a rat model of intestinal ischemia-reperfusion injury and the underlying mechanism. METHODS: A total of 66 rats were divided into three dietary groups (lipid-free, soybean oil, and fish oil groups). Each animal was administered total parenteral nutrition for 3 days, followed by induction of intestinal ischemia for 100 min. Animals subjected to sham surgery served as the controls. Intestinal tissue and blood were harvested 6 and 12 h after the surgery, then, assessment of the histological damage score, plasma-related parameters, and statistical evaluation were performed. RESULTS: The histological damage score in the intestinal tissues was significantly lower in the fish oil group than in the soybean oil group (P = 0.0121). The late-phase urinary level of 8-hydroxy-2-deoxyguanosine was also significantly lower in the fish oil group as compared with that in the other groups (P = 0.0267). Furthermore, the plasma level of high-mobility group box 1 protein was also significantly lower in the fish oil group as compared with that in the lipid-free group (P = 0.0398). CONCLUSION: It appeared that intravenous administration of an omega-3 lipid emulsion prior to ischemia-reperfusion injury reduced the oxidative stress and severity of tissue damage. Modification of membrane fatty acids may serve as the mechanism underlying this reduction of tissue damage.
Asunto(s)
Emulsiones Grasas Intravenosas/administración & dosificación , Ácidos Grasos Omega-3/administración & dosificación , Intestino Delgado/irrigación sanguínea , Estrés Oxidativo/efectos de los fármacos , Nutrición Parenteral Total , Daño por Reperfusión/terapia , Animales , Modelos Animales de Enfermedad , Masculino , Ratas , Ratas Wistar , Daño por Reperfusión/metabolismoRESUMEN
AIM: To investigate the hepatoprotective effect of MK615, a Japanese apricot extract, in an animal model, and its clinical therapeutic effect. METHODS: Wistar rats were administered physiological saline (4 mL/kg) or MK615 solution (4 mL/kg) for 7 d. On the sixth d, acute hepatic injury was induced by administering a single intraperitoneal injection (ip) of D-galactosamine hydrochloride (D-GalN) (600 mg/kg). Plasma levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were determined, and liver tissues were used for histopathological analysis. Fifty-eight patients with liver disorders [hepatitis C (n = 40), non-alcoholic fatty liver disease (n = 15), and autoimmune liver disease (n = 3)] were orally administered commercially available Misatol ME-containing MK615 (13 g/d) daily for 12 wk. Blood and urine were sampled immediately before and 6 wk, 12 wk, and 16 wk after the start of intake to measure various biochemical parameters. The percentage change in ALT and AST levels after 12 wk from the pre-intake baseline served as a primary endpoint. RESULTS: D-GalN effectively induced acute hepatic injury in the rats. At 48 h after the ip injection of D-GalN, the plasma levels of ALT (475.6 ± 191.5 IU/L vs 225.3 ± 194.2 IU/L, P < 0.05) and AST (1253.9 ± 223.4 IU/L vs 621.9 ± 478.2 IU/L, P < 0.05) in the MK615 group were significantly lower than the control group. Scattered single cell necrosis, loss of hepatocytes, and extensive inflammatory cell infiltration were observed in hepatic tissue samples collected from the control group. However, these findings were less pronounced in the group receiving MK615. At the end of the clinical study, serum ALT and AST levels were significantly decreased compared with pre-intake baseline levels from 103.5 ± 58.8 IU/L to 71.8 ± 39.3 IU/L (P < 0.05) and from 93.5 ± 55.6 IU/L to 65.5 ± 34.8 IU/L (P < 0.05), respectively. A reduction of ≥ 30% from the pre-study baseline ALT level was observed in 26 (45%) of the 58 patients, while 25 (43%) patients exhibited similar AST level reductions. The chronic hepatitis C group exhibited significant ALT and AST level reductions from 93.4 ± 51.1 IU/L to 64.6 ± 35.1 IU/L (P < 0.05) and from 94.2 ± 55.5 IU/L to 67.2 ± 35.6 IU/L (P < 0.05), respectively. A reduction of ≥ 30% from the pre-study baseline ALT level was observed in 20 (50%) of the 40 patients. ALT levels in both the combined ursodeoxycholic acid (UDCA) treatment and the UDCA uncombined groups were significantly lower after Misatol ME administration. MK615 protected hepatocytes from D-GalN-induced cytotoxicity in rats. Misatol ME decreased elevated ALT and AST levels in patients with liver disorders. CONCLUSION: These results suggest that MK615 and Misatol ME are promising hepatoprotective agents for patients with liver disorders.
Asunto(s)
Hígado Graso/tratamiento farmacológico , Hepatitis C/tratamiento farmacológico , Hepatitis Autoinmune/tratamiento farmacológico , Extractos Vegetales/uso terapéutico , Prunus , Adulto , Anciano , Anciano de 80 o más Años , Alanina Transaminasa/metabolismo , Animales , Aspartato Aminotransferasas/metabolismo , Enfermedad Hepática Inducida por Sustancias y Drogas/tratamiento farmacológico , Enfermedad Hepática Inducida por Sustancias y Drogas/metabolismo , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Modelos Animales de Enfermedad , Hígado Graso/metabolismo , Hígado Graso/patología , Femenino , Galactosamina/efectos adversos , Hepatitis C/metabolismo , Hepatitis C/patología , Hepatitis Autoinmune/metabolismo , Hepatitis Autoinmune/patología , Humanos , Hígado/enzimología , Hígado/patología , Masculino , Persona de Mediana Edad , Enfermedad del Hígado Graso no Alcohólico , Ratas , Ratas Wistar , Resultado del TratamientoRESUMEN
The Japanese apricot, a commonly consumed food called 'Ume' in Japan, has been used for a traditional Japanese medicine for centuries. MK615, an extract of compounds from 'Ume', has strong antitumorigenic and antiinflammatory effects including the induction of apoptosis and autophagy, and inhibition of cytokine production mediated via the inhibition of MAPKs signaling including ERK-1/2, JNK and p38MAPK. The inhibitor of DNA binding 1 (Id-1), a basic helix-loop-helix (bHLH) transcription factor family, is essential for DNA binding and the transcriptional regulation of various proteins that play important roles in the development, progression and invasion of tumors. In melanoma, Id-1 is constitutively expressed in the late and early stages, suggesting it as a therapeutic target in patients with melanoma. This study reports that MK615 profoundly reduced both the mRNA- and protein expression levels of Id-1 and inhibited cell growth in A375 melanoma cells. MK615 markedly inhibited the phosphorylation of ERK1/2, which is associated with Id-1 protein expression in A375 cells. Id-1-specific RNAi induced the death of A375 cells. Moreover, the expression of Bcl-2 was decreased by both MK615 and Id-1-specific RNAi in A375 cells. The results suggest that MK615 is a potential therapeutic agent for treating malignant melanoma.
Asunto(s)
Regulación Neoplásica de la Expresión Génica , Proteína 1 Inhibidora de la Diferenciación/metabolismo , Sistema de Señalización de MAP Quinasas , Melanoma/patología , Extractos Vegetales/farmacología , Prunus/química , Antineoplásicos Fitogénicos/farmacología , Apoptosis , Muerte Celular , Línea Celular Tumoral , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Proteína 1 Inhibidora de la Diferenciación/genética , Melanoma/genética , Melanoma/metabolismo , Fosforilación , Extractos Vegetales/química , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Interferencia de ARN , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/genética , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
Tobacco-smoke exposure is linked to carcinogenic, oxidative and inflammatory cellular reactions. Green tea has been reported to have anti-release properties against various pro-inflammatory cytokines. To determine the effects of green tea extract (GTE) on serum high mobility group box-1 (HMGB1) levels in rats exposed to cigarette smoke (CS), we divided rats into 4 treatment groups: (1) CS only, (2) dietary supplement with GTE (3 mg/d) and CS (GCS1), (3) dietary supplement with GTE (4.5 mg/d) and CS (GCS2) and (4) a control group. HMGB1 and cotinine serum levels were analyzed by ELISA. The average serum HMGB1 level in the CS group was significantly higher than the other groups (p < 0.01), indicating the release of HMGB1 into the blood was stimulated by CS exposure, while GTE consumption suppressed HMGB1 levels. Rats exposed to CS had an average serum cotinine level of 37 ng/ml, indicating tobacco related compounds were present in the rats' blood. However, treatment with GTE did not reduce cotinine levels in all groups. Cotinine stimulated HMGB1 secretion in a dose- and time-dependent manner, and HMGB1 levels were suppressed by GTE in murine macrophage cell lines. Our results show GTE supplementation may offer beneficial systemic effects and suppress HMGB1 by protecting against cell inflammation.
Asunto(s)
Antiinflamatorios/farmacología , Proteína HMGB1/antagonistas & inhibidores , Extractos Vegetales/farmacología , Té , Contaminación por Humo de Tabaco/efectos adversos , Animales , Supervivencia Celular , Cotinina/sangre , Relación Dosis-Respuesta a Droga , Masculino , Ratas , Factores de TiempoRESUMEN
The Japanese apricot, known as Ume in Japanese, has been a traditional Japanese medicine for centuries, and is a familiar and commonly consumed food. The health benefits of Ume are now being widely recognized and have been strengthened by recent studies showing that MK615, an extract of compounds from Ume, has strong anticancer and anti-inflammatory effects. However, the potential role of MK615 in the periodontal field remains unknown. Here, we found that MK615 significantly reduced the production of pro-inflammatory mediators (tumor necrosis factor-alpha and interleukin-6) induced by Porphyromonas gingivalis lipopolysaccharide (LPS), a major etiological agent in localized chronic periodontitis, in murine macrophage-like RAW264.7 cells. MK615 markedly inhibited the phosphorylation of ERK1/2, p38MAPK, and JNK, which is associated with pro-inflammatory mediator release pathways. Moreover, MK615 completely blocked LPS-triggered NF-kappaB activation. The present results suggest that MK615 has potential as a therapeutic agent for treating inflammatory diseases such as periodontitis.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Interleucina-6/antagonistas & inhibidores , Macrófagos/efectos de los fármacos , Quinasas de Proteína Quinasa Activadas por Mitógenos/antagonistas & inhibidores , Extractos Vegetales/farmacología , Porphyromonas gingivalis , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Transporte Activo de Núcleo Celular , Animales , Antiinflamatorios no Esteroideos/uso terapéutico , Línea Celular , Interleucina-6/biosíntesis , Lipopolisacáridos/inmunología , Macrófagos/enzimología , Ratones , Periodontitis/tratamiento farmacológico , Periodontitis/microbiología , Extractos Vegetales/uso terapéutico , Factor de Transcripción ReIA/metabolismo , Factor de Necrosis Tumoral alfa/biosíntesisRESUMEN
High mobility group box-1 protein (HMGB1), primarily from the nucleus, is released into the extracellular milieu either passively from necrotic cells or actively through secretion by monocytes/macrophages. Extracellular HMGB1 acts as a potent inflammatory agent by promoting the release of cytokines such as tumor necrosis factor (TNF)-alpha, has procoagulant activity, and is involved in death due to sepsis. Accordingly, HMGB1 is an appropriate therapeutic target. In this study, we found that an extract of Prunus mume Sieb. et Zucc. (Ume) fruit (Ume extract), an abundant source of triterpenoids, strongly inhibited HMGB1 release from lipopolysaccharide (LPS)-stimulated macrophage-like RAW264.7 cells. The inhibitory effect on HMGB1 release was enhanced by authentic oleanolic acid (OA), a naturally occurring triterpenoid. Similarly, the HMGB1 release inhibitor in Ume extract was found to be OA. Regarding the mechanisms of the inhibition of HMGB1 release, the OA or Ume extract was found to activate the transcription factor Nrf2, which binds to the antioxidative responsive element, and subsequently the heme oxygenase (HO)-1 protein was induced, indicating that the inhibition of HMGB1 release from LPS-stimulated RAW264.7 cells was mediated via the Nrf2/HO-1 system; an essentially antioxidant effect. These results suggested that natural sources of triterpenoids warrant further evaluation as 'rescue' therapeutics for sepsis and other potentially fatal systemic inflammatory disorders.
Asunto(s)
Proteína HMGB1/metabolismo , Macrófagos/efectos de los fármacos , Ácido Oleanólico/farmacología , Prunus/química , Vías Secretoras/efectos de los fármacos , Animales , Antioxidantes/aislamiento & purificación , Antioxidantes/farmacología , Línea Celular , Evaluación Preclínica de Medicamentos , Hemo-Oxigenasa 1/metabolismo , Lipopolisacáridos/farmacología , Macrófagos/metabolismo , Ratones , Factor 2 Relacionado con NF-E2/metabolismo , Ácido Oleanólico/aislamiento & purificación , Extractos Vegetales/química , Extractos Vegetales/farmacología , Transporte de Proteínas/efectos de los fármacosRESUMEN
OBJECTIVE: Tissue hypoxia is closely associated with arthritis pathogenesis, and extracellular high mobility group box chromosomal protein 1 (HMGB-1) released from injured cells also has a role in arthritis development. This study was thus undertaken to investigate the hypothesis that extracellular HMGB-1 may be a coupling factor between hypoxia and inflammation in arthritis. METHODS: Concentrations of tumor necrosis factor alpha, interleukin-6, vascular endothelial growth factor, lactic acid, lactate dehydrogenase, and HMGB-1 were measured in synovial fluid (SF) samples from patients with inflammatory arthropathy (rheumatoid arthritis and pseudogout) and patients with noninflammatory arthropathy (osteoarthritis). The localization of tissue hypoxia and HMGB-1 was also examined in animal models of collagen-induced arthritis (CIA). In cell-based experiments, the effects of hypoxia on HMGB-1 release and its associated cellular events (i.e., protein distribution and cell viability) were studied. RESULTS: In SF samples from patients with HMGB-1-associated inflammatory arthropathy (i.e., samples with HMGB-1 levels >2 SD above the mean level in samples from patients with noninflammatory arthropathy), concentrations of HMGB-1 were significantly correlated with those of lactic acid, a marker of tissue hypoxia. In CIA models in which the pathologic phenotype could be attenuated by HMGB-1 neutralization, colocalization of HMGB-1 with tissue hypoxia in arthritis lesions was also observed. In cell-based experiments, hypoxia induced significantly increased levels of extracellular HMGB-1 by the cellular processes of secretion and/or apoptosis-associated release, which was much more prominent than the protein release in necrotic cell injury potentiated by oxidative stress. CONCLUSION: These findings indicate that tissue hypoxia and its resultant extracellular HMGB-1 might play an important role in the development of arthritis.
Asunto(s)
Artritis/metabolismo , Proteína HMGB1/análisis , Hipoxia/metabolismo , Inflamación/metabolismo , Articulaciones/metabolismo , Líquido Sinovial/química , Adulto , Anciano , Anciano de 80 o más Años , Animales , Artritis/patología , Artritis Experimental/inducido químicamente , Artritis Experimental/metabolismo , Artritis Experimental/patología , Western Blotting , Células Cultivadas , Femenino , Técnica del Anticuerpo Fluorescente , Humanos , Hipoxia/patología , Inflamación/patología , Interleucina-1/análisis , L-Lactato Deshidrogenasa/análisis , Ácido Láctico/análisis , Masculino , Ratones , Persona de Mediana Edad , Ratas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Índice de Severidad de la Enfermedad , Membrana Sinovial/metabolismo , Membrana Sinovial/patología , Factor de Necrosis Tumoral alfa/análisis , Factor A de Crecimiento Endotelial Vascular/análisisRESUMEN
Bofutsushosan (BOF), a traditional Chinese formulation (Kampo formulation in Japanese), is widely used for patients with obesity and hyperlipidemia resulting from long-term inappropriate lifestyles. Since atherosclerosis, a lifestyle-related disease, is accompanied by an abnormal accumulation of vascular smooth muscle cells (VSMCs) in the intimal area of the artery, we investigated the preventive effect of BOF on intimal thickening. Oral administration of BOF extracts 3 d before and 7 d after balloon endothelial denudation dose dependently suppressed the intimal thickening and proliferation of VSMCs in the intimal area in rat carotid arteries. This model has a similar pathologic process to atherosclerosis and is considered to be an "accelerated atherosclerosis" model. BOF extract also dose dependently inhibited the migration of cultured VSMCs. BOF extract suppressed serum lipid levels, which are a major risk factor for atherosclerosis. These findings clarified the usefulness of BOF in cardiovascular risk-reduction therapy.
Asunto(s)
Vasos Sanguíneos/patología , Fármacos Cardiovasculares/farmacología , Medicamentos Herbarios Chinos/farmacología , Endotelio Vascular/fisiología , Músculo Liso Vascular/citología , Animales , Aorta Torácica/citología , Aorta Torácica/efectos de los fármacos , Vasos Sanguíneos/efectos de los fármacos , Peso Corporal/efectos de los fármacos , Arterias Carótidas/efectos de los fármacos , Arterias Carótidas/patología , Cateterismo , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Ingestión de Alimentos/efectos de los fármacos , Lípidos/sangre , Masculino , Músculo Liso Vascular/efectos de los fármacos , RatasRESUMEN
Intimal formation of animal carotid arteries induced by balloon endothelial denudation has been considered to be an "accelerated atherosclerosis" model and used in primary screening methods to evaluate natural drugs and chemical candidates. The aim of the present study was to examine whether intimal formation is prevented by Bezoar Bovis (dried cattle gallbladder stones: Niuhuang in Chinese and Go-o in Japanese), which has been used to prevent heart palpitation in patients with hypertension. The intimal-to-medial area ratio in rat carotid arteries 7 days after balloon endothelial denudation was significantly reduced by oral administration of Bezoar Bovis. Bezoar Bovis also suppressed vascular smooth muscle cells (VSMCs) proliferation, which is thought to play important roles in the intimal formation after endothelial damage and also atherosclerosis resulting from long-term inappropriate lifestyle. The present findings suggest that Bezoar Bovis may be useful for preventing atherosclerosis and for protection against restenosis after percutaneous coronary intervention, for which effective reduction method is not currently available.
Asunto(s)
Proliferación Celular/efectos de los fármacos , Músculo Liso Vascular/citología , Túnica Íntima/efectos de los fármacos , Túnica Media/efectos de los fármacos , Animales , Arterias Carótidas/patología , Bovinos , Modelos Animales de Enfermedad , Cálculos Biliares , Masculino , Medicina Tradicional China , Músculo Liso Vascular/efectos de los fármacos , Inhibidores de Agregación Plaquetaria/farmacología , Ratas , Ratas Wistar , Túnica Íntima/lesiones , Túnica Íntima/patología , Túnica Media/patología , ortoaminobenzoatos/farmacologíaRESUMEN
Oral administration of Saiko-ka-Ryukotsu-Borei-To (SRB), a traditional Chinese formulation, has been found to prevent intimal thickening of the carotid artery after balloon endothelial denudation in cholesterol-fed rats. To clarify the mechanism of this effect, the present study investigated if SRB inhibits vascular smooth muscle cell (VSMC) migration, which plays an important role in the development of intimal thickening after endothelial injury. The serum (SRB serum) sampled from cholesterol-fed rats treated orally with SRB for 3 days before and 4 days after the injury dose-dependently inhibited the migration of cultured VSMCs. When added directly to cultured VSMCs, the SRB extract did not inhibit VSMC migration. It is remarkable that SRB serum, which may contain a much lower concentration of SRB ingredients compared with the SRB extract, inhibited cultured VSMC migration. The present testing system using serum obtained from animals treated orally with traditional Chinese formulations may be useful for clarifying the pharmacological efficacy of such drugs, including many non-absorbable components. Furthermore, it may be useful in the search for new active compounds in serum after oral administration of traditional Chinese formulations, the active metabolites of which have not been identified.
Asunto(s)
Movimiento Celular/efectos de los fármacos , Medicamentos Herbarios Chinos/farmacología , Músculo Liso Vascular/efectos de los fármacos , Administración Oral , Animales , Arterias Carótidas/patología , Cateterismo , Células Cultivadas , Colesterol en la Dieta , Medios de Cultivo Condicionados , Inmunohistoquímica , Masculino , Músculo Liso Vascular/patología , Ratas , Ratas Wistar , Suero , Túnica Íntima/lesiones , Túnica Íntima/patologíaRESUMEN
It has been suggested that oxidation of low-density lipoprotein (LDL) plays an important role in the initiation and progression of atherosclerosis. In the present study, we determined the anti-atherogenic effects of egg yolk-enriched garlic powder (EGP), which has been used as a traditional health-promoting food in southern Japan since ancient times, on LDL oxidation and oxidant stress-induced cell injury models. We confirmed that EGP inhibits copper-induced LDL oxidation in a dose-dependent manner. We also observed that pretreatment of EGP significantly suppressed the production of peroxides in HL60 cells and protected endothelial cells from hydrogen peroxide-induced cell injury. These findings might, in part, be ascribed to the biodistribution of garlic compounds and egg yolk interaction, and suggest that EGP might be useful in the prevention of atherosclerosis.
Asunto(s)
Yema de Huevo/metabolismo , Ajo/metabolismo , Lipoproteínas LDL/metabolismo , Estrés Oxidativo/fisiología , Arteriosclerosis/metabolismo , Arteriosclerosis/prevención & control , Disponibilidad Biológica , Supervivencia Celular/fisiología , Células Endoteliales/metabolismo , Células HL-60 , Humanos , Japón , Oxidación-Reducción , Especies Reactivas de Oxígeno/metabolismo , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismoRESUMEN
We examined the inhibitory effects of traditional Chinese formulations (TCFs: Kampo formulation in Japanese) on intimal thickening of the carotid artery injured by balloon endothelial denudation in rats. Among the eight TCFs examined oren-gedoku-to (huanglian-jiedu-tang in Chinese), choto-san (diao-teng-san), saiko-ka-ryukotsu-borei-to (chaihujia-longgu-muli-tang) and dai-joki-to (da-cheng-qi-tang) significantly inhibited the intimal thickening 7 days after denudation. These four TCFs also inhibited proliferation of vascular smooth muscle cells (VSMC), which may play a central role in the development of restenosis after balloon endothelial denudation. The present results suggest that further evaluation of these four TCFs as inhibitors of VSMC proliferation to prevent arteriosclerosis is warranted.
Asunto(s)
Traumatismos de las Arterias Carótidas/tratamiento farmacológico , Medicamentos Herbarios Chinos/uso terapéutico , Endotelio Vascular/lesiones , Animales , Antiinflamatorios no Esteroideos/uso terapéutico , Peso Corporal/fisiología , Estenosis Carotídea/tratamiento farmacológico , Estenosis Carotídea/patología , Cateterismo , División Celular/efectos de los fármacos , Indicadores y Reactivos , Luminol , Masculino , Músculo Liso Vascular/citología , Músculo Liso Vascular/efectos de los fármacos , Antígeno Nuclear de Célula en Proliferación , Ratas , Ratas Wistar , ortoaminobenzoatos/uso terapéuticoRESUMEN
Oral administration of Saiko-ka-Ryukotsu-Borei-To (SRB), a traditional Chinese formulation, dose dependently inhibited intimal thickening in carotid artery injured by balloon endothelial denudation in cholesterol-fed rats. SRB also inhibited vascular smooth muscle cell (VSMC) proliferation, which is assessed by counting the VSMCs immunoreactive with antiproliferating cell nuclear antigen (PCNA) antibody in the intimal area. VSMC proliferation is considered to play a central role in the development of intimal thickening. SRB slightly, but not significantly, reduced serum total cholesterol and low-density lipoprotein cholesterol. These results indicate that the suppressive effect of SRB on intimal thickening may result from its inhibitory effect against VSMC proliferation, but does not depend on lowering of lipid levels. The balloon injury model used in this study has similar pathological processes to restenosis after percutaneous coronary intervention (PCI). Therefore the present results may provide a new therapeutic strategy using SRB to reduce restenosis after PCI in the treatment of patients with ischemic coronary artery disease. Furthermore, since it is considered that artery restenosis after balloon injury in PCI is "accelerated atherosclerosis, " SRB may have beneficial effects in atherosclerosis that develops over a long clinical course in hyperlipidemia, diabetes, etc.
Asunto(s)
Arterias Carótidas/efectos de los fármacos , Cateterismo/efectos adversos , Colesterol en la Dieta/efectos adversos , Medicamentos Herbarios Chinos/farmacología , Endotelio Vascular/efectos de los fármacos , Animales , Arterias Carótidas/patología , Medicamentos Herbarios Chinos/uso terapéutico , Endotelio Vascular/patología , Masculino , Músculo Liso Vascular/efectos de los fármacos , Músculo Liso Vascular/patología , Ratas , Ratas Wistar , Túnica Íntima/efectos de los fármacos , Túnica Íntima/patologíaRESUMEN
We report here that the traditional Chinese formulation, Chaihu-jia-Longgu-Muli-tang (CLM), significantly inhibited the increase in intimal thickening in rat carotid artery injured by balloon endothelial denudation, which mimics many aspects of restenosis after percutaneous coronary intervention (PCI) in humans. CLM, Saiko-ka-Ryukotsu-Borei-to in Japanese, is commonly prescribed for symptoms accompanying hypertension and atherosclerosis in Japanese Kampo medical care. CLM administered orally 1 week before and 1, 4 and 8 weeks after balloon injury inhibited the increase in intimal area, intimal/medial ratio and stenosis ratio. To our knowledge, this is the first report demonstrating inhibitory effects of a traditional Chinese formulation on intimal thickening of carotid artery after balloon injury. It is worth noting that CLM maintained its inhibitory effect up to 8 weeks after balloon injury. The reduction in intimal thickening by CLM could have resulted from inhibition of intimal smooth muscle cell proliferation, which was assessed by immuno-histochemical analysis using monoclonal antibody against proliferating cell nuclear antigen. Therefore, CLM may be a favourable candidate for prevention of restenosis after PCI. Moreover CLM may have a therapeutic value in the prevention of atherosclerosis, because restenosis after PCI is considered to be an accelerated atherosclerosis.
Asunto(s)
Angioplastia Coronaria con Balón/efectos adversos , Traumatismos de las Arterias Carótidas/tratamiento farmacológico , Estenosis Carotídea/prevención & control , Medicamentos Herbarios Chinos/farmacología , Endotelio Vascular/efectos de los fármacos , Túnica Íntima/patología , Animales , Peso Corporal/efectos de los fármacos , Traumatismos de las Arterias Carótidas/etiología , División Celular/efectos de los fármacos , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/uso terapéutico , Ingestión de Alimentos/efectos de los fármacos , Endotelio Vascular/lesiones , Endotelio Vascular/patología , Humanos , Hiperplasia/tratamiento farmacológico , Hiperplasia/prevención & control , Inmunohistoquímica , Masculino , Medicina Tradicional China , Medicina Kampo , Estructura Molecular , Músculo Liso Vascular/citología , Músculo Liso Vascular/efectos de los fármacos , Músculo Liso Vascular/metabolismo , Inhibidores de Agregación Plaquetaria/farmacología , Inhibidores de Agregación Plaquetaria/uso terapéutico , Complicaciones Posoperatorias/tratamiento farmacológico , Complicaciones Posoperatorias/prevención & control , Antígeno Nuclear de Célula en Proliferación/análisis , Ratas , Ratas Wistar , Túnica Íntima/efectos de los fármacos , Túnica Íntima/lesiones , ortoaminobenzoatos/farmacología , ortoaminobenzoatos/uso terapéuticoRESUMEN
The antioxidant effects of 1,5-anhydro-D-fructose (1,5-AF), a unique anhydrohexulose, were studied in 1,1-diphenyl-2-picrylhydrazyl (DPPH) solution, in human cells along with lipid peroxidation of low-density lipoprotein (LDL). We have confirmed that 1,5-AF scavenges DPPH radicals directly in solution and inhibits the formation of hydrogen peroxide and superoxide anion, typical reactive oxygen species (ROS), induced by phorbol myristate acetate (PMA) in a dose-dependent manner in THP-1 cells. We also observed the dose-dependent antioxidant effects of 1,5-AF on copper-mediated LDL oxidation. These findings suggest that 1,5-AF might play a role in reducing the risk of atherosclerosis and may help prevent coronary heart disease.