RESUMO
This study was conducted to assess the effect of Evodiae Fructus 70% ethanol extract (EFE) on the pathology of atopic dermatitis using in vitro and in vivo models. The major compounds in EFE were identified by ultra-performance liquid chromatography with tandem mass spectrometry as rutaecarpine, evodiamine, evodol, dehydroevodiamine, limonin, synephrine, evocarpine, dihydroevocarpine, and hydroxyevodiamine. EFE significantly decreased chemokine levels in tumor necrosis factor-α/interferon-γ-stimulated HaCaT cells. In house dust mite-treated NC/Nga mice, topical application of EFE significantly decreased the dermatitis score, epidermal hyperplasia and thickening, mast cell infiltration, and plasma levels of histamine and corticosterone. Thymic stromal lymphopoietin, CD4+ T cells, interleukin-4, and intercellular adhesion molecule-1 expression in the lesioned skin was reduced in the treated mice. The mechanism of EFE was elucidated using transcriptome analysis, followed by experimental validation using Western blotting in HaCaT cells. EFE down-regulated the activation of Janus kinase (JAK)-signal transducers and activators of transcription (STAT) and mitogen-activated protein kinases (MAPK) signaling pathways in HaCaT cells. EFE improves atopic dermatitis-like symptoms by suppressing inflammatory mediators, cytokines, and chemokines by regulating the JAK-STAT and MAPK signaling pathways, suggesting its use as a potential agent for the treatment of atopic dermatitis.
Assuntos
Dermatite Atópica , Evodia , Camundongos , Animais , Humanos , Dermatite Atópica/patologia , Pyroglyphidae , Evodia/metabolismo , Células HaCaT , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Citocinas/metabolismo , Quimiocinas/metabolismo , Dermatophagoides pteronyssinus , Etanol/farmacologia , Pele/metabolismoRESUMO
Daeshiho-tang (DSHT), a traditional herbal formula with diverse pharmacological effects, has shown promise in medicine owing to its anti-hypertensive, anti-diabetic, and anti-inflammatory properties. However, the precise molecular mechanism underlying these effects remains unclear. Thus, we investigated the effect of DSHT on inflammatory response and oxidative stress to understand its molecular mechanism using lipopolysaccharide (LPS)-induced macrophage (RAW 264.7) cells. DSHT decreased the contents of nitric oxide (NO) and prostaglandin E2 (PGE2) through downregulating inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein expressions. DSHT suppressed the LPS-induced TLR4 as well as MyD88, subsequently suppressing the NF-κB activation and the phosphorylation of MAPK (p38, ERK, and JNK). Radical scavenging activity results revealed a dose-dependent response of DSHT with diminished ABTS activity, a hallmark of oxidative stress potential. Furthermore, DSHT enhanced Nrf2 and HO-1 expression in response to LPS. Collectively, our findings indicated that DSHT exert anti-inflammatory effect and regulating oxidative stress by modulating TLR4/MyD88, NF-κB, MAPK, and Nrf2/HO-1 pathways, consequently can provide potential therapeutic strategy for the prevention and treatment of inflammation and oxidative stress-related diseases.
Assuntos
Lipopolissacarídeos , NF-kappa B , Animais , Camundongos , NF-kappa B/metabolismo , Lipopolissacarídeos/toxicidade , Lipopolissacarídeos/metabolismo , Fator 88 de Diferenciação Mieloide/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Receptor 4 Toll-Like/metabolismo , Macrófagos/metabolismo , Anti-Inflamatórios/uso terapêutico , Células RAW 264.7 , Estresse Oxidativo , Óxido Nítrico Sintase Tipo II/metabolismo , Ciclo-Oxigenase 2/metabolismoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Traditional herbal medicines have diverse efficacy and are increasingly used worldwide. However, some of these herbal medicines have toxicities or side effects, but the scientific understanding of traditional herbal medicine toxicity has not yet been established. Asiasari Radix et Rhizoma (ARE) is known as a herbal medicine used to relieve pain, and recent studies have shown that ARE has anticancer and antimelanogenesis efficacy. AIM OF THE STUDY: Current study was conducted to assess the potential genotoxicity of an ethanolic extract of ARE. MATERIALS AND METHODS: The genotoxixity of ARE was confirmed by the bacterial reverse mutation assay (Ames test), a mammalian chromosomal aberration test, and a micronucleus test in vivo using ICR mice and comet assay using Sprague-Dawley rats. RESULTS: ARE showed no genotoxicity in a micronucleus test up to 2000 mg/kg body weight in vivo. By contrast, the chromosomal aberration test showed that ARE induced an increase in the number of chromosomal aberrations after treatment for 6 h with a metabolic activation system and for 6 and 22 h without the metabolic activation system when compared with vehicle control. In the Ames test, all strains except TA1535, with or without a metabolic activation system, showed an increase in the number of revertant mutant colonies in the ARE-treated group. In comet assay, DNA damage was observed in the stomach when ARE was administered. CONCLUSION: ARE potentially shows genotoxicity by inducing DNA damage.
Assuntos
Aristolochiaceae/química , Dano ao DNA , Medicamentos de Ervas Chinesas/toxicidade , Animais , Bactérias/efeitos dos fármacos , Bactérias/genética , Peso Corporal/efeitos dos fármacos , Aberrações Cromossômicas/induzido quimicamente , Ensaio Cometa , Cricetulus , Etanol , Fígado/efeitos dos fármacos , Masculino , Camundongos Endogâmicos ICR , Testes para Micronúcleos , Testes de Mutagenicidade , Ratos Sprague-Dawley , Estômago/efeitos dos fármacosRESUMO
BACKGROUND: There are various Helicobacter species colonizing the stomachs of animals. Although Helicobacter species usually cause asymptomatic infection in the hosts, clinical signs can occur due to gastritis associated with Helicobacter in animals. Among them, Helicobacter pylori is strongly associated with chronic gastritis, gastric ulcers, and gastric cancers. As the standard therapies used to treat H. pylori have proven insufficient, alternative options are needed to prevent and eradicate the diseases associated with this bacterium. Cheonwangbosim-dan (CBD), a traditional herbal formula that is popular in East Asia, has been commonly used for arterial or auricular flutter, neurosis, insomnia, and cardiac malfunction-induced disease. OBJECTIVES: The present study investigated the antimicrobial effect of CBD on H. pylori-infected human gastric carcinoma AGS cells and model mice. METHODS: AGS cells were infected with H. pylori and treated with a variety of concentrations of CBD or antibiotics. Mice were given 3 oral inoculations with H. pylori and then dosed with CBD (100 or 500 mg/kg) for 4 weeks or with standard antibiotics for 1 week. One week after the last treatment, gastric samples were collected and examined by histopathological analysis, real-time quantitative polymerase chain reaction, and immunoblotting. RESULTS: Our results showed that CBD treatment of AGS cells significantly reduced the H. pylori-induced elevations of interleukin-8, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). In the animal model, CBD treatment inhibited the colonization of H. pylori and the levels of malondialdehyde, inflammation, proinflammatory cytokines, iNOS, and COX-2 in gastric tissues. CBD also decreased the phosphorylation levels of p38 mitogen-activated protein kinase family. CONCLUSIONS: This study suggests that CBD might be a prospective candidate for treating H. pylori-induced gastric injury.
Assuntos
Antibacterianos/farmacologia , Anti-Inflamatórios/farmacologia , Gastrite/tratamento farmacológico , Infecções por Helicobacter/tratamento farmacológico , Helicobacter pylori/efeitos dos fármacos , Animais , Carcinoma , Linhagem Celular Tumoral , Masculino , Medicina Tradicional Coreana , Camundongos , Camundongos Endogâmicos C57BL , Neoplasias GástricasRESUMO
Atopic dermatitis (AD) is a prevalent chronic inflammatory skin disease. The use of immunomodulatory corticosteroids in AD treatment causes adverse side effects. Therefore, novel natural anti-inflammatory therapeutics are needed. The aim of the present study was to investigate the anti-allergic and anti-inflammatory activities of kuwanon G and morusin. To investigate the effect of kuwanon G and morusin on skin inflammation, enzyme-linked immunosorbent assays (ELISA) to quantitate secreted (RANTES/CCL5), thymus- and activation-regulated chemokine (TARC/CCL17), and macrophage-derived chemokine (MDC/CCL22) were performed, followed by Western blotting to measure the phosphorylation of signal transducer and activator of transcription 1 (STAT1) and nuclear transcription factor-κB (NF-κB) p65 in tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ)-stimulated HaCaT keratinocytes. In order to evaluate the anti-allergic effects, ELISA to quantify histamine and leukotriene C4 (LTC4) production and Western blotting to measure 5-lipoxygenase (5-LO) activation were performed using PMA and A23187-stimulated MC/9 mast cells. Kuwanon G reduced the release of RANTES/CCL5, TARC/CCL17, and MDC/CCL22 via down-regulation of STAT1 and NF-κB p65 signaling in TNF-α and IFN-γ-stimulated HaCaT keratinocytes. Kuwanon G also inhibited histamine production and 5-LO activation in PMA and A23187-stimulated MC/9 mast cells. Morusin inhibited RANTES/CCL5 and TARC/CCL17 secretion via the suppression of STAT1 and NF-κB p65 phosphorylation in TNF-α and IFN-γ-stimulated HaCaT keratinocytes, and the release of histamine and LTC4 by suppressing 5-LO activation in PMA and A23187-stimulated MC/9 mast cells. Kuwanon G and morusin are potential anti-inflammatory mediators for the treatment of allergic and inflammatory skin diseases such as AD.
Assuntos
Antialérgicos/farmacologia , Anti-Inflamatórios/farmacologia , Flavonoides/farmacologia , Queratinócitos/efeitos dos fármacos , Queratinócitos/metabolismo , Mastócitos/efeitos dos fármacos , Mastócitos/metabolismo , Antialérgicos/química , Anti-Inflamatórios/química , Biomarcadores , Linhagem Celular , Quimiocinas/metabolismo , Cromatografia Líquida de Alta Pressão , Flavonoides/química , Humanos , Estrutura Molecular , NF-kappa B/metabolismo , Fosforilação , Fator de Transcrição STAT1/metabolismoRESUMO
In the present study, Bojungikgi-tang (BJIKT: Buzhongyiqi-tang, Hochuekki-to) and Palmijihwang-hwan (PMJHH: Baweidìhuang-wan, Hachimijio-gan), traditional herbal formulas, investigated anti-inflammatory efficacies in murine macrophage cell line and the influence on the activities of drug-metabolizing enzymes (DMEs). The anti-inflammatory potentials of the herbal formulas were evaluated to inhibit the production of the inflammatory mediators and cytokines and the protein expression of inducible nitric oxide and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-treated RAW 264.7 cells. The activities of the major human DMEs, cytochrome P450 isozymes (CYP450s) and UDP-glucuronosyltransferase isozymes (UGTs), were measured by in vitro enzyme assay systems. BJIKT and PMJHH significantly suppressed the prostaglandin E2 (PGE2) production (IC50 = 317.3 and 282.2 µg/mL, respectively) and the protein expression of COX-2 in LPS-treated RAW264.7 cells. On the human microsomal DMEs, BJIKT inhibited the activities of CYP1A2 (IC50 = 535.05 µg/mL), CYP2B6 (IC50 > 1000 µg/mL), CYP2C9 (IC50 = 800.78 µg/mL), CYP2C19 (IC50 = 563.11 µg/mL), CYP2D6 (IC50 > 1000 µg/mL), CYP2E1 (IC50 > 1000 µg/mL), CYP3A4 (IC50 = 879.60 µg/mL), UGT1A1 (IC50 > 1000 µg/mL), and UGT1A4 (IC50 > 1000 µg/mL), but it showed no inhibition of the UGT2B7 activity at doses less than 1000 µg/mL. PMJHH inhibited the CYP2D6 activity (IC50 = 280.89 µg/mL), but IC50 values of PMJHH exceeded 1000 µg/mL on the activities of CYP1A2, CYP2C19, CYP2E1, and CYP3A4. At concentrations less than 1000 µg/mL, PMJHH did not affect the activities of CYP2B6, CYP2C9, UGT1A1, UGT1A4, and UGT2B7. The results indicate that both BJIKT and PMJHH may be potential candidates to prevent and treat PGE2- and COX-2-mediated inflammatory diseases. In addition, this study will expand current knowledge about herb-drug interactions by BJIKT and PMJHH.
Assuntos
Anti-Inflamatórios/farmacologia , Inibidores Enzimáticos/farmacologia , Macrófagos/efeitos dos fármacos , Microssomos Hepáticos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Animais , Anti-Inflamatórios/química , Inibidores das Enzimas do Citocromo P-450/farmacologia , Sistema Enzimático do Citocromo P-450/metabolismo , Composição de Medicamentos , Inibidores Enzimáticos/química , Glucuronosiltransferase/antagonistas & inibidores , Glucuronosiltransferase/metabolismo , Humanos , Macrófagos/enzimologia , Macrófagos/imunologia , Camundongos , Microssomos Hepáticos/enzimologia , Microssomos Hepáticos/imunologia , Extratos Vegetais/química , Células RAW 264.7RESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Traditional herbal formula Gyejibokryeong-hwan (GJBRH; Guizhifuling-wan, Keishibukuryo-gan) consisting five medicinal herbs has been used to treat uterine disorders, gynecological diseases and blood stasis syndrome in Asia. AIM OF THE STUDY: We evaluated the safety of GJBRH in Crl:CD Sprague-Dawley (SD) rats over a period of 13 weeks. MATERIALS AND METHODS: To confirm the stability of the components of GJBRH, we analyzed the component contents in GJBRH at different storage periods, using high-performance liquid chromatography. Male and female SD rats were orally administered with GJBRH at doses of 0, 1000, 2000 and 5000â¯mg/kg/day for 13 weeks and assessed after a 4-week recovery period. Mortality, changes in body weight and food consumption, organ weights, hematology and serum biochemistry were monitored during the experimental period, along with clinical observations, ophthalmological examinations, urinalysis and histopathology. RESULTS: There were no significant differences among the eight marker compounds in GJBRH according to storage period. No significant GJBRH-treatment-related toxicological changes were observed in mortality or ophthalmological examinations in either sex. However, soft feces were observed in the male 5000â¯mg/kg/day group. In addition, there were significant changes in body weight and food consumption in both male and female rats treated with GJBRH at a dose of 5000â¯mg/kg/day. In the hematological examinations, we found a significant increase in white blood cells, neutrophils and fibrinogen in the 5000â¯mg/kg/day groups. In the urinalysis, a decrease in the total protein and albumin and an increase in the ovalbumin/globulin ratio were observed in both male and female rats treated with GJBRH at a dose of 5000â¯mg/kg/day. Histopathological examinations revealed erosion/ulcers and dilated glands in the stomachs of males from the 5000â¯mg/kg/day group, and squamous cell hyperplasia and epithelial atrophy was observed in the stomachs of both male and female rats treated with GJBRH at a dose of 5000â¯mg/kg/day. CONCLUSION: The no-observed-adverse-effect level (NOAEL) was 2000â¯mg/kg/day for both sexes.
Assuntos
Medicamentos de Ervas Chinesas/toxicidade , Animais , Atrofia/induzido quimicamente , Contagem de Células Sanguíneas , Peso Corporal/efeitos dos fármacos , Medula Óssea/efeitos dos fármacos , Medula Óssea/patologia , Medicamentos de Ervas Chinesas/análise , Ingestão de Alimentos/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/patologia , Feminino , Hiperplasia/induzido quimicamente , Fígado/efeitos dos fármacos , Fígado/patologia , Masculino , Nível de Efeito Adverso não Observado , Tamanho do Órgão/efeitos dos fármacos , Compostos Fitoquímicos/análise , Compostos Fitoquímicos/toxicidade , Ratos Sprague-Dawley , Baço/efeitos dos fármacos , Baço/patologia , Estômago/efeitos dos fármacos , Estômago/patologia , Testes de Toxicidade SubcrônicaRESUMO
Macrophages are phagocytes that play important roles in health and diseases. Acyl-CoA:cholesterol acyltransferase 1 (ACAT1) converts cellular cholesterol to cholesteryl esters and is expressed in many cell types. Unlike global Acat1 knockout (KO), myeloid-specific Acat1 KO ( Acat1-) does not cause overt abnormalities in mice. Here, we performed analyses in age- and sex-matched Acat1-M/-M and wild-type mice on chow or Western diet and discovered that Acat1-M/-M mice exhibit resistance to Western diet-induced obesity. On both chow and Western diets, Acat1-M/-M mice display decreased adipocyte size and increased insulin sensitivity. When fed with Western diet, Acat1-M/-M mice contain fewer infiltrating macrophages in white adipose tissue (WAT), with significantly diminished inflammatory phenotype. Without Acat1, the Ly6Chi monocytes express reduced levels of integrin-ß1, which plays a key role in the interaction between monocytes and the inflamed endothelium. Adoptive transfer experiment showed that the appearance of leukocytes from Acat1-M/-M mice to the inflamed WAT of wild-type mice is significantly diminished. Under Western diet, Acat1-M/-M causes suppression of multiple proinflammatory genes in WAT. Cell culture experiments show that in RAW 264.7 macrophages, inhibiting ACAT1 with a small-molecule ACAT1-specific inhibitor reduces inflammatory responses to lipopolysaccharide. We conclude that under Western diet, blocking ACAT1 in macrophages attenuates inflammation in WAT. Other results show that Acat1-M/-M does not compromise antiviral immune response. Our work reveals that blocking ACAT1 suppresses diet-induced obesity in part by slowing down monocyte infiltration to WAT as well as by reducing the inflammatory responses of adipose tissue macrophages.
Assuntos
Acetil-CoA C-Acetiltransferase/genética , Acetil-CoA C-Acetiltransferase/fisiologia , Dieta , Inflamação/genética , Inflamação/patologia , Resistência à Insulina/genética , Macrófagos/patologia , Obesidade/genética , Esterol O-Aciltransferase/genética , Esterol O-Aciltransferase/fisiologia , Adipócitos/patologia , Tecido Adiposo Branco/metabolismo , Tecido Adiposo Branco/patologia , Animais , Tamanho Celular , Feminino , Intolerância à Glucose/genética , Intolerância à Glucose/metabolismo , Inflamação/imunologia , Integrina beta1/metabolismo , Macrófagos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Obesidade/fisiopatologia , Células RAW 264.7RESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Helicobacter pylori, which is found in the stomachs of approximately half of the world's population, has been associated with the development of chronic gastritis and gastric cancer. Hwanglyeonhaedok-tang (HHT) is a popular traditional medicine for the therapies of gastric ulcers and gastritis. AIM OF THE STUDY: The emerging resistance of H. pylori to antibiotics arouses requirement on alternative nonantibiotic-based therapies. In the present study, we investigated the anti-inflammatory activity and anti-microbial activity of HHT against H. pylori in vitro and in an H. pylori-infected mouse model. MATERIALS AND METHODS: H. pylori were treated with various concentrations of HHT and then incubated with human gastric carcinoma AGS cells. For the in vivo study, mice were orally infected with H. pylori three times over the course of 1 week, and then subjected to daily administration of HHT (120 or 600â¯mg/kg) for 4 weeks or standard triple therapy for 1 week. At the scheduled termination of the experiment, all mice were killed and their stomachs were collected for histological examination, quantitative real-time PCR, and Western blot analysis. RESULTS: Our in vitro studies showed that HHT treatment inhibited the adhesion of H. pylori to AGS cells and suppressed the H. pylori-induced increases of inflammatory regulators, such as interleukin (IL)-8, cyclooxygenase 2 (COX-2), and inducible nitric oxide synthase (iNOS). In the mouse model, HHT treatment significantly reduced H. pylori colonization, inflammation, and the levels of IL-1ß, IL-6, C-X-C motif chemokine ligand 1 (CXCL1), tumor necrosis factor alpha (TNF-α), COX-2, and iNOS in gastric mucosa. Further investigation showed that HHT treatment reduced the H. pylori-induced phosphorylations of p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinases 1/2 (ERK1/2), c-Jun N-terminal protein kinase (JNK), and nuclear factor-kappa B (NF-κB). CONCLUSIONS: Our findings collectively suggest that HHT has anti-inflammatory activity and antibacterial activity against H. pylori and could be an alternative to antibiotics for preventing H. pylori infection.
Assuntos
Antibacterianos/farmacologia , Anti-Inflamatórios/farmacologia , Gastrite/prevenção & controle , Infecções por Helicobacter/tratamento farmacológico , Helicobacter pylori/efeitos dos fármacos , Extratos Vegetais/farmacologia , Estômago/efeitos dos fármacos , Animais , Aderência Bacteriana/efeitos dos fármacos , Linhagem Celular Tumoral , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Mucosa Gástrica/metabolismo , Gastrite/metabolismo , Gastrite/microbiologia , Gastrite/patologia , Infecções por Helicobacter/metabolismo , Infecções por Helicobacter/microbiologia , Infecções por Helicobacter/patologia , Helicobacter pylori/patogenicidade , Interações Hospedeiro-Patógeno , Humanos , Mediadores da Inflamação/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fosforilação , Estômago/microbiologia , Estômago/patologiaRESUMO
BACKGROUND: Gyejibokryeong-hwan (Guizhi Fuling Wan in China), a mixture of five herbal plants, is a well-known treatment for renal diseases including those associated with climacteric syndrome. However, the genotoxicity of Gyejibokryeong-hwan has not yet been well established. METHODS: The present study investigated that the genotoxicity of an aqueous extract of Gyejibokryeong-hwan (GJBRHE): an in vitro chromosomal aberration test using Chinese hamster lung cells, an in vitro bacterial reverse mutation assay (Ames test) with Salmonella typhimurium and Escherichia coli strains, and an in vivo micronucleus test using ICR mouse bone marrow. RESULTS: GJBRHE with or without the S9 mix showed no genotoxicity in the Ames test up to 5000 µg/plate or in the in vivo MN test up to 2000 mg/kg body weight. In contrast, the chromosomal aberration test showed that GJBRHE induced an increase in the number of chromosomal aberrations compared with the control after treatment for 6 h with 4200 µg/mL GJBRHE in the presence of the S9 mix and for 22 h with 800 µg/mL GJBRHE in the absence of the S9 mix. CONCLUSIONS: GJBRHE did not cause detectable genotoxic effects in the bacterial mutation test or the in vivo MN test, however genotoxic effect was detected in the in vitro chromosomal aberration assay. Our results suggest that GJBRHE may be associated with a low risk of carcinogenesis. Thus, further detailed experiments would be needed to clarify the compound responsible for inducing this genotoxicity of GJBRHE and to determine its mechanism.
Assuntos
Aberrações Cromossômicas/efeitos dos fármacos , Dano ao DNA/efeitos dos fármacos , Medicamentos de Ervas Chinesas/toxicidade , Mutagênicos/toxicidade , Animais , Peso Corporal/efeitos dos fármacos , Medula Óssea/efeitos dos fármacos , Linhagem Celular , Cricetinae , Escherichia coli/efeitos dos fármacos , Camundongos , Testes para Micronúcleos , Testes de Mutagenicidade , Salmonella typhimurium/efeitos dos fármacosRESUMO
OBJECTIVE: Brown and white adipose tissue exerts pleiotropic effects on systemic energy metabolism in part by releasing endocrine factors. Neuregulin 4 (Nrg4) was recently identified as a brown fat-enriched secreted factor that ameliorates diet-induced metabolic disorders, including insulin resistance and hepatic steatosis. However, the physiological mechanisms through which Nrg4 regulates energy balance and glucose and lipid metabolism remain incompletely understood. The aims of the current study were: i) to investigate the regulation of adipose Nrg4 expression during obesity and the physiological signals involved, ii) to elucidate the mechanisms underlying Nrg4 regulation of energy balance and glucose and lipid metabolism, and iii) to explore whether Nrg4 regulates adipose tissue secretome gene expression and adipokine secretion. METHODS: We examined the correlation of adipose Nrg4 expression with obesity in a cohort of diet-induced obese mice and investigated the upstream signals that regulate Nrg4 expression. We performed metabolic cage and hyperinsulinemic-euglycemic clamp studies in Nrg4 transgenic mice to dissect the metabolic pathways regulated by Nrg4. We investigated how Nrg4 regulates hepatic lipid metabolism in the fasting state and explored the effects of Nrg4 on adipose tissue gene expression, particularly those encoding secreted factors. RESULTS: Adipose Nrg4 expression is inversely correlated with adiposity and regulated by pro-inflammatory and anti-inflammatory signaling. Transgenic expression of Nrg4 increases energy expenditure and augments whole body glucose metabolism. Nrg4 protects mice from diet-induced hepatic steatosis in part through activation of hepatic fatty acid oxidation and ketogenesis. Finally, Nrg4 promotes a healthy adipokine profile during obesity. CONCLUSIONS: Nrg4 exerts pleiotropic beneficial effects on energy balance and glucose and lipid metabolism to ameliorate obesity-associated metabolic disorders. Biologic therapeutics based on Nrg4 may improve both type 2 diabetes and non-alcoholic fatty liver disease (NAFLD) in patients.
Assuntos
Adipocinas/sangue , Ácidos Graxos/metabolismo , Neurregulinas/metabolismo , Hepatopatia Gordurosa não Alcoólica/metabolismo , Obesidade/metabolismo , Adipócitos/metabolismo , Animais , Células Cultivadas , Dieta Hiperlipídica/efeitos adversos , Metabolismo Energético , Glucose/metabolismo , Fígado/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Neurregulinas/genética , Hepatopatia Gordurosa não Alcoólica/etiologia , Obesidade/etiologiaRESUMO
The aim of the present study was to investigate whether Syzygium aromaticum water extract (SAWE) has a protective effect against ethanolinduced gastric injury in rats. Acute gastric injury was induced via intragastric administration of absolute ethanol at a dose of 5 ml/kg. SAWE (250 or 500 mg/kg/day) or cimetidine (100 mg/kg/day), which was used as a positive control, were administered to the rats 2 h prior to ethanol administration for 3 days. All rats were sacrificed 24 h following the final ethanol administration. To examine whether SAWE has a gastroprotective effect, assays were performed to assess the contents of malondialdehyde (MDA) and glutathione (GSH), the activities of catalase, glutathioneStransferase and superoxide dismutase, and an immune-linked immunosorbent assay was performed for prostaglandin E2 (PGE2) production in gastric tissues by hematoxylin and eosin and periodic acid-Schiff staining. Histological assessment of the gastric wall was performed. Compared with ethanol treatment alone, treatment with SAWE at a dose of 250 mg/kg/day significantly decreased the gastric MDA content and increased the GSH content, catalase activity, and production of gastric PGE2. Histological assessment showed that SAWE attenuated inflammatory cell infiltration and the loss of epithelial cells. These findings suggested that SAWE protected against ethanolinduced gastric mucosal injury in the rats. These effects appeared to be associated with antioxidant activity, activation of the production of PGE2, suppression of inflammatory cell infiltration and loss of epithelial cells in the gastric mucosa. Collectively, SAWE may be beneficial in the prevention of gastric disease associated to oxidative stress.
Assuntos
Antioxidantes/farmacologia , Etanol/efeitos adversos , Gastrite/etiologia , Gastrite/metabolismo , Extratos Vegetais/farmacologia , Syzygium/química , Animais , Catalase/metabolismo , Dinoprostona/metabolismo , Modelos Animais de Doenças , Ativação Enzimática/efeitos dos fármacos , Mucosa Gástrica/efeitos dos fármacos , Mucosa Gástrica/metabolismo , Mucosa Gástrica/patologia , Gastrite/tratamento farmacológico , Gastrite/patologia , Glutationa/metabolismo , Mediadores da Inflamação/metabolismo , Peroxidação de Lipídeos , Masculino , Oxirredução/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Ratos , Superóxido Dismutase/metabolismoRESUMO
Gleditsia sinensis thorns have traditionally been used to treat edema and carbuncles and drain abscesses. In the present study, a simultaneous analysis of four flavonoids [(+)catechin, ()epicatechin, eriodictyol and quercetin] and two phenolic compounds (caffeic acid and ethyl gallate), obtained from a 70% ethanol extract of G. sinensis, was performed using highperformance liquid chromatographyphotodiode array techniques. In addition, the inhibitory activities of the solvent fractions from a G. sinensis extract and its major constituents on the lipopolysaccharidestimulated production of inflammatory mediators by macrophage RAW 264.7 cells and the tumor necrosis factor (TNF)α and interferon (IFN)γ (TI)stimulated production of chemokines by HaCaT keratinocyte cells were investigated. The established analytical method showed high linearity, with a correlation coefficient of ≥0.9998. The limits of detection and quantification of the six compounds were 0.0370.425 and 0.1241.418 µg/ml, respectively. The ethyl acetate fraction inhibited nitric oxide and prostaglandin E2 production in RAW 264.7 cells and the production of thymus and activationregulated chemokine (TARC) in HaCaT cells more than did the other fractions. Furthermore, the six compounds reduced the production of TARC, macrophagederived chemokine and regulated on activation normal Tcell expressed and secreted in TIstimulated HaCaT cells; in particular, ethyl gallate and quercetin exhibited a significant dosedependent inhibition. Further elucidation of the signaling pathways involved in the Thelper cell 2 chemokine inhibition by G. sinensis is necessary to facilitate the design of therapeutic agents for the inflammatory response.
Assuntos
Anti-Inflamatórios/farmacologia , Gleditsia/química , Queratinócitos/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Animais , Linhagem Celular , Quimiocina CCL17/biossíntese , Dinoprostona/biossíntese , Avaliação Pré-Clínica de Medicamentos , Humanos , Queratinócitos/imunologia , Queratinócitos/metabolismo , Limite de Detecção , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos , Óxido Nítrico/biossíntese , Caules de Planta/químicaRESUMO
Traditional herbal formulas made from Scutellariae Radix (SR), the root of Scutellaria baicalensis, have previously been used in the treatment of inflammatory diseases, such as atherosclerosis. The aim of the present study was to investigate the effects of SR on low-density lipoprotein (LDL) oxidation and inflammation in macrophages, which are early events in the development of atherosclerosis. High-performance liquid chromatography photo-diode array analysis was used to obtain a three-dimensional chromatogram of SR. The antioxidative effects of SR were evaluated by determining its scavenging activities against ABTS and DPPH radicals. The inhibitory effect of SR on LDL oxidation was examined using a thiobarbituric acid-reactive substance assay and a relative electrophoretic mobility assay. In addition, the anti-inflammatory effects of SR were evaluated in lipopolysaccharide (LPS)-induced RAW264.7 murine macrophage cells. The results showed that SR exhibited radical-scavenging activities in a dose-dependent manner; in addition, SR attenuated the Cu2+-induced oxidation of LDL as well as significantly inhibited nitric oxide (NO) production and inducible NO synthase (iNOS) expression in LPS-induced RAW264.7 cells. Furthermore, SR induced the protein expression of heme oxygenase-1 (HO-1) in RAW264.7 cells. In conclusion, the results of the present study demonstrated that SR decreased the oxidation of LDL and suppressed inflammatory responses in macrophages, which occurred at least in part via the induction of HO1. These results therefore suggested that SR may be a potential therapeutic agent for the treatment of atherosclerosis.
Assuntos
Aterosclerose/tratamento farmacológico , Heme Oxigenase-1/biossíntese , Inflamação/tratamento farmacológico , Lipoproteínas LDL/antagonistas & inibidores , Extratos Vegetais/administração & dosagem , Animais , Aterosclerose/metabolismo , Aterosclerose/patologia , Heme Oxigenase-1/genética , Humanos , Inflamação/induzido quimicamente , Inflamação/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Lipoproteínas LDL/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase Tipo II/biossíntese , Oxirredução , Extratos Vegetais/química , Scutellaria baicalensis/químicaRESUMO
CONTEXT: The fruit pulp of Luffa cylindrica Roemer (Cucurbitaceae) (LC) has been used to induce hemostasis, resolve phlegm and clear fever in traditional Korean medicine. However, the efficacy of LC has not been examined in atopic dermatitis (AD). OBJECTIVE: A 70% ethanol extract of LC was evaluated to determine anti-inflammation and anti-AD effects in vitro and in vivo. MATERIALS AND METHODS: The inhibitory effects of LC on the production of PGE2 and histamine were respectively measured in lipopolysaccharide-treated (1 µg/mL) RAW264.7 macrophages and phorbol-12 myristate 13-acetate (50 nM) and A23187 (1 µM)-stimulated HMC-1 mast cells. The production of AD-related chemokines (RANTES, TARC, and MDC) were evaluated in IFN-γ and TNF-α-stimulated (10 ng/mL, each) HaCaT keratinocytes. LC (10 mg/mouse/d) was topically applied to the dorsal skin and ears of Dermatophagoides farina (Pyroglyphidae)-sensitized Nc/Nga mice for 4 weeks. RESULTS: The IC50 values of LC on PGE2 and histamine production were 16.89 and 139.9 µg/mL, individually. The production of TARC and RANTES were inhibited 20% and 12% by LC (50 µg/mL) in HaCaT cells, respectively (p < 0.05). In sensitized-NC/Nga mice, the plasma levels of IgE and histamine were suppressed 36% and 41% by LC, respectively (p < 0.05). LC also reduced hemorrhage, hypertrophy, and hyperkeratosis of the epidermis and infiltration of mast cells in the dorsal skin and ear. DISCUSSION AND CONCLUSION: LC can inhibit AD-like skin lesions and reduce the generation of IgE via inhibition of the inflammatory responses. LC has potential as a therapeutic agent to treat allergic diseases, including AD.
Assuntos
Dermatite Atópica/prevenção & controle , Dermatophagoides farinae/imunologia , Luffa/química , Extratos Vegetais/uso terapêutico , Animais , Linhagem Celular , Quimiocinas/imunologia , Dermatite Atópica/sangue , Dermatite Atópica/imunologia , Dermatite Atópica/patologia , Dinoprostona/biossíntese , Frutas/química , Liberação de Histamina/efeitos dos fármacos , Humanos , Imunoglobulina E/sangue , Queratinócitos/efeitos dos fármacos , Queratinócitos/imunologia , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Masculino , Mastócitos/efeitos dos fármacos , Mastócitos/imunologia , Camundongos Endogâmicos , Óxido Nítrico/biossíntese , Extratos Vegetais/isolamento & purificaçãoRESUMO
The aim of this study was to explore the protective effects of Trichosanthes kirilowii ethanol extract (TKE) against cisplatin-induced acute renal failure (ARF). In the in vitro study, TKE-pretreated porcine kidney cells (PK15) exhibited enhanced cell viability after cisplatin (15 µg mL(- 1)) treatment in both MTT and crystal violet assays. PK15 cells pretreated with TKE (50 µg mL(- 1)) exhibited increased glutathione content, decreased reactive oxygen species production and ameliorated p53 expression. In vivo study, rats were administered with TKE for 4 weeks before cisplatin (5 mg kg(- 1)) injection. TKE (100 mg kg(- 1)) decreased blood urea nitrogen and creatinine levels by 24% and 47%, respectively, in comparison with cisplatin-alone group. In addition, TKE pretreatment ameliorated cisplatin-induced oxidative stress, as evidenced by increased antioxidative enzyme levels and decreased lipid peroxidation levels. Moreover, TKE pretreatment reduced histopathological alterations in the kidney with decreased apoptotic cells. Taken together, TKE might be beneficial in treating cisplatin-induced ARF.
Assuntos
Injúria Renal Aguda/tratamento farmacológico , Cisplatino/efeitos adversos , Rim/patologia , Extratos Vegetais/farmacologia , Trichosanthes/química , Injúria Renal Aguda/induzido quimicamente , Animais , Apoptose/efeitos dos fármacos , Nitrogênio da Ureia Sanguínea , Células Cultivadas , Creatinina/sangue , Glutationa/metabolismo , Glutationa Peroxidase/metabolismo , Rim/efeitos dos fármacos , Peroxidação de Lipídeos/efeitos dos fármacos , Malondialdeído/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Ratos , Espécies Reativas de Oxigênio/metabolismo , SuínosRESUMO
We investigated the protective effects against acute renal failure (ARF) of Anemarrhena asphodeloides (AA) and performed simultaneous analysis of three compounds, neomangiferin (1), mangiferin (2), and isomangiferin (3) in AA using a high-performance liquid chromatography-photodiode array. To measure the protective effect of ARF, the levels of reactive oxygen species (ROS) and glutathione depletion were determined using a kit. HPLC analysis was performed using a Gemini Cia column at 40 degrees C. The mobile phase used gradient elution with 1.0% (v/v) aqueous acetic acid (A) and 1.0% (v/v) acetic acid in acetonitrile (B). The flow rate was 1.0 mL/min. In our assay, AA extract inhibits cisplatin-induced production of intracellular ROS. Pre-incubation of AA extracts (10-200 microg/mL) markedly maintained cell viability compared with controls in the noncisplatin-treated cells. Calibration curves of all compounds showed good linearity (r2 > or = 0.9992). Recoveries of the three compounds were 98.9-103.4%. The relative standard deviations of intra- and inter-day precision were 0.07-1.73% and 0.12-1.49%, respectively. The amounts of the three components were 1.22-20.63 mg/g. The AA extract has potential as a therapeutic agent for treatment of ARF. In addition, the established method will help to improve quality control of AA.
Assuntos
Anemarrhena/química , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão/métodos , Extratos Vegetais/farmacologia , Insuficiência Renal/prevenção & controle , Animais , Linhagem Celular , Cisplatino/toxicidade , Glutationa/metabolismo , Fitoterapia , Extratos Vegetais/química , Raízes de Plantas/química , Plantas Medicinais , Espécies Reativas de Oxigênio/metabolismoRESUMO
Saussurea lappa is a traditional herbal medicine used for to treat various inflammatory diseases. In this study, we investigated the protective effects of S. lappa against atopic dermatitis using human keratinocyte HaCaT cells, murine mast cell line MC/9 cells, and a house dust mite-induced atopic dermatitis model of Nc/Nga mice. Treatment with the S. lappa caused a significant reduction in the mRNA levels and production of inflammatory chemokines and cytokine, including thymus- and activation-regulated chemokine (TARC), macrophage-derived chemokine (MDC), regulated on activation, normal T-cell expressed and secreted (RANTES), and interleukin-8 (IL-8) in tumor necrosis factor-α/interferone-γ-stimulated HaCaT cells. S. lappa exhibited the significant reduction in histamine production in MC/9 cells. In the atopic dermatitis model, S. lappa significantly reduced the dermatitis score and serum IgE and TARC levels. In addition, the back skin and ears of S. lappa-treated Nc/Nga mice exhibited reduced histological manifestations of atopic skin lesions such as erosion, hyperplasia of the epidermis and dermis, and inflammatory cell infiltration. In conclusion, an extract of S. lappa effectively suppressed the development of atopic dermatitis, which was closely related to the reduction of chemokines and cytokine. Our study suggests that S. lappa may be a potential treatment for atopic dermatitis.
Assuntos
Quimiocinas/antagonistas & inibidores , Dermatite Atópica/metabolismo , Mediadores da Inflamação/antagonistas & inibidores , Ácaros , Extratos Vegetais/farmacologia , Saussurea/química , Animais , Sequência de Bases , Linhagem Celular , Quimiocinas/biossíntese , Primers do DNA , Liberação de Histamina/efeitos dos fármacos , Humanos , Imunoglobulina E/sangue , Mediadores da Inflamação/metabolismo , Camundongos , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
OBJECTIVE: Pinellia ternata is an important plant in traditional Chinese medicine. This study describes the anti-inflammatory effects of a water extract of P. ternata (PTE) in allergic airway inflammation in a model of asthma in mice. MATERIALS AND METHODS: BALB/c mice were sensitized with ovalbumin (OVA) and, upon an OVA aerosol challenge, developed airway eosinophilia, mucus hypersecretion, elevations in cytokine, chemokine, and immunoglobulin levels and overexpression of inducible nitric oxide (iNOS). RESULTS: Intragastric administration of PTE significantly attenuated OVA-induced influx of total leukocytes, eosinophils, neutrophils, macrophages and lymphocytes into lungs, and attenuated levels of interleukin (IL)-4, IL-13 and tumor necrosis factor-α (TNF-α), in a dose-dependent manner. PTE also significantly reduced the plasma levels of total and OVA-specific immunoglobulin (Ig)E release into the airspace. Histological studies showed that PTE inhibited OVA-induced lung tissue eosinophilia and airway mucus production. Moreover, in whole lung tissue lysates, immunohistology showed that PTE markedly attenuated the OVA-induced increase in mucin 5AC and iNOS expression. CONCLUSIONS: These results indicate that PTE has protective effects against allergic airway inflammation.
Assuntos
Antiasmáticos/uso terapêutico , Asma/prevenção & controle , Medicamentos de Ervas Chinesas/uso terapêutico , Muco/metabolismo , Pinellia/química , Sistema Respiratório/efeitos dos fármacos , Animais , Antiasmáticos/administração & dosagem , Antiasmáticos/isolamento & purificação , Asma/imunologia , Asma/patologia , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Contagem de Células , Citocinas/imunologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/isolamento & purificação , Feminino , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Inflamação/imunologia , Inflamação/patologia , Inflamação/prevenção & controle , Camundongos , Camundongos Endogâmicos BALB C , Muco/imunologia , Ovalbumina/imunologia , Sistema Respiratório/imunologia , Sistema Respiratório/patologiaRESUMO
Amomum compactum is commonly used in Korean traditional medicine. In this study, we demonstrate that A. compactum ethanolic extract (ACEE) has anti-inflammatory effects in a lipopolysaccharide-induced RAW 264.7 cell model of inflammation. In this system, ACEE prominently inhibited the production of nitric oxide (NO), prostaglandin E(2) (PGE(2)), interleukin (IL)-6 and tumor necrosis factor (TNF)-α, and inhibited the protein expression of inducible nitric oxide synthase and cyclooxygenase-2. Furthermore, ACEE treatment inhibited the translocation of nuclear factor-kappaB (NF-κB) and the degradation of inhibitory factor-kappaB alpha, but enhanced the expression of heme oxygenase (HO)-1 and the nuclear translocation of nuclear factor-erythroid 2 (Nrf2). Treatment with tin protoporphyrin IX dichloride (SnPP), a selective HO-1 inhibitor, reversed the ACEE-induced suppression of NO production, suggesting that the induction of HO-1 is involved in the suppression of NO, TNF-α, and IL-6 production by ACEE. Taken together, these results suggest that ACEE have anti-inflammatory effects occurring through HO-1 induction, which leads to suppression of the blocking NF-κB.