RESUMO
Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by skin barrier dysfunction. Sargassum horneri (S. horneri) is a brown alga that has been widely used in traditional medicine of eastern Asian countries. Recent studies proved that a brown alga S. horneri has anti-inflammatory activity. In this study, we investigated the effect of S. horneri ethanol extract (SHE) against AD in 2,4-dinitrobenzene (DNCB) induced AD in NC/Nga mice. We observed that SHE treatment decreased the epidermal thickness and epidermal hyperplasia that had been worsened through DNCB application. Moreover, SHE significantly inhibited the proliferation of mast cells and decreased the expression of IL-13 on CD4⺠cells prompted by elevated thymic stromal lymphopoietin (TSLP) expression in DNCB-induced AD in mice. We also demonstrated that SHE directly inhibited the expression of keratinocyte-produced TSLP known to exacerbate skin barrier impairment. Especially, the decrease of filaggrin, an integral component of proper skin barrier function through a function in aggregating keratin filaments, observed in DNCB-induced AD mice was significantly improved when treated with SHE. More importantly, we proved that SHE was able to decrease the serum levels of IgG1 and IgG2â, two crucial factors of AD, indicating the protective effect of SHE. Taken together, our findings suggest that SHE may protect NC/Nga mice against DNCB-induced AD via promoting skin barrier function.
Assuntos
Dermatite Atópica , Extratos Vegetais , Sargassum , Dermatopatias , Animais , Camundongos , Anti-Inflamatórios/uso terapêutico , Citocinas/metabolismo , Dermatite Atópica/induzido quimicamente , Dermatite Atópica/tratamento farmacológico , Dinitrobenzenos/efeitos adversos , Imunoglobulina G , Interleucina-13/metabolismo , Queratinas/metabolismo , Extratos Vegetais/farmacologia , Polifenóis/farmacologia , Sargassum/química , Pele/metabolismo , Dermatopatias/induzido quimicamente , Dermatopatias/tratamento farmacológicoRESUMO
The present study evaluated the anti-obesity effect of sulforaphane (SFN) and glucoraphanin (GRN) in broccoli leaf extract (BLE) on 3T3-L1 adipocytes and ob/ob mice. Based on Oil Red O staining and triglyceride (TG) assay, SFN and BLE significantly reduced (P<.05) both lipid accumulation and TG content in the differentiated 3T3-L1 adipocytes. SFN and BLE increased 2-NBDG uptake by 3T3-L1 adipocytes in a dose-dependent manner. Western blot analysis confirmed that SFN and BLE increased the phosphorylation levels of both AMPK (Thr172) and ACC (Ser79), and reduced the expression of HMGCR in liver and white adipose tissues of ob/ob mice. Histological analysis revealed that SFN and BLE ameliorated hepatic steatosis, and reduced the size of adipocyte in ob/ob mice. Treatment with SFN and BLE significantly reduced (P<.05) TG content, low-density lipoprotein (LDL) cholesterol, total cholesterol (TC), and glucose in the serum of ob/ob mice. RNA sequencing analysis showed that up- or down-regulation of 32 genes related to lipid metabolism was restored to control level in both SFN and BLE-treated ob/ob mice groups. A protein-protein interaction (PPI) network was constructed via STRING analysis, and Srebf2, Pla2g2c, Elovl5, Plb1, Ctp1a, Lipin1, Fgfr1, and Plcg1 were located in the functional hubs of the PPI network of lipid metabolism. Overall results suggest that the SFN content in BLE exerts a potential anti-obesity effect by normalizing the expression of genes related to lipid metabolism, which are up- or down-regulated in ob/ob mice.
Assuntos
Adipócitos/metabolismo , Fármacos Antiobesidade/farmacologia , Brassica/química , Isotiocianatos/farmacologia , Obesidade/metabolismo , Extratos Vegetais/farmacologia , Sulfóxidos/farmacologia , Células 3T3-L1 , Proteínas Quinases Ativadas por AMP/metabolismo , Adipócitos/citologia , Adipócitos/efeitos dos fármacos , Adipócitos Brancos/citologia , Animais , Glicemia/metabolismo , Glucose/metabolismo , Glucosinolatos/farmacologia , Metabolismo dos Lipídeos/efeitos dos fármacos , Lipídeos/sangue , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Camundongos , Camundongos Obesos , Obesidade/tratamento farmacológico , Obesidade/patologia , Oximas/farmacologia , Fosforilação , Folhas de Planta/química , Transcriptoma , Triglicerídeos/metabolismoRESUMO
CONTEXT: Sargassum horneri (Turner) C. Agardh (Sargassaceae) is a brown marine alga used in oriental medicine to treat allergic conditions. OBJECTIVE: This study clarifies the effect of polyphenol-containing S. horneri ethanol extract (SHE) on T-helper type-2 (Th2) polarisation. MATERIALS AND METHODS: All mice (BALB/c mice, n = 12) except in the healthy control group were first sensitised with an intraperitoneal injection of ovalbumin (OVA; 20 µg) and alum (2 mg) on Day 0 and Day 14. Similarly, phosphate-buffered saline (PBS) was injected according to the same schedule into the healthy control mice. After the final administration, splenocytes were obtained. OVA sensitised mice were challenged with OVA (100 µg/mL) in the absence or presence (62.5 and 125 µg/mL) of SHE while healthy control group remained untreated. RESULTS: SHE (0-1000 µg/mL) was not cytotoxic to splenocytes and demonstrated IC50 values of 3.27 and 3.92 mg/mL, respectively, at 24 and 48 h of incubation. SHE suppressed cell proliferation at concentrations ≥62.5 µg/mL. SHE treatment (125 µg/mL) subdued (by 1.8-fold) the population expansion of CD3+CD4+ helper T cells induced by OVA challenge. SHE attenuated the OVA-induced activation of respective transcription factors GATA3 and NLRP3. Simultaneously, highly elevated levels of cytokines interleukin (IL)-4 and IL-5 caused by OVA stimulation were removed completely and IL-13 suppressed by 1.5-fold. CONCLUSIONS: SHE exhibits Th2 immune suppression under OVA stimulation via GATA3- and NLRP3-dependent IL-4, IL-5, and IL-13 suppression. Therefore, SHE could be therapeutically useful for alleviating the symptoms of allergen-mediated immune diseases.
Assuntos
Extratos Vegetais/farmacologia , Polifenóis/farmacologia , Sargassum/química , Células Th2/imunologia , Animais , Citocinas/imunologia , Relação Dose-Resposta a Droga , Fator de Transcrição GATA3/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Ovalbumina , Extratos Vegetais/administração & dosagem , Polifenóis/administração & dosagem , Polifenóis/isolamento & purificação , Fator de Transcrição STAT5/metabolismo , Baço/citologia , Baço/efeitos dos fármacos , Baço/imunologiaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Sargassum horneri (Turner) C. Agardh is well known in East Asia as an edible brown alga rich in bioactive compounds. It has an ethnopharmacological significance in traditional Chinese medicine to treat inflammatory disorders varying from edema, furuncles, dysuria to cardiovascular diseases. AIM OF THE STUDY: Surge of fine dust (FD), in densely populated areas, have been reported to cause adverse health conditions ranging from respiratory diseases to inflammatory skin disorders. The current study investigates the protective effects of an ethanol extract from S. horneri (SHE) on FD-induced inflammatory responses and impaired skin hydration in HaCaT keratinocytes. MATERIALS AND METHODS: Intracellular reactive oxygen species (ROS) generation was evaluated with the 2',7'-Dichlorofluorescin diacetate (DCFH-DA) stain. Anti-inflammatory properties of SHE in FD-stimulated HaCaT keratinocytes were investigated for the suppression of nuclear factor (NF)-κB and mitogen-activated protein kinase (MAPK) pathways and downregulation of pro-inflammatory cytokines. As a means of studying FD-induced skin barrier disruption and the effects of SHE on stratum corneum hydration-controlling factors, tight junction regulatory mediators, and hyaluronic acid (HA) production were evaluated using keratinocytes. RESULTS: SHE suppressed the intracellular ROS production, simultaneously improving cell viability in FD-stimulated keratinocytes. Also, SHE upregulated anti-inflammatory cytokine interleukin (IL)-4 while downregulating inflammatory cytokines IL-1ß, IL-6, IL-8, tumor necrosis factor (TNF)-α; epidermal and epithelial cytokines IL-25, IL-33, and thymic stromal lymphopoietin (TSLP); thymus and activation-regulated chemokine (TARC), macrophage-derived chemokine (MDC) and regulated upon activation, normally T-expressed, and presumably secreted expression and suppressed (RANTES) chemokine, MAPK and NF-κB mediators in a dose-dependent manner. Furthermore, SHE ameliorated filaggrin, involucrin, lymphoepithelial Kazal-type-related inhibitor (LEKTI), signifying its beneficial effects on deteriorated skin hydration caused by FD-induced inflammation. SHE further exhibited its skin protective effects regulating the tight junction proteins; Occludin, zonula occludens (ZO)-1, claudin-1, claudin-4, claudin-7, and claudin-23 while increasing the production of HA minimizing skin damage. CONCLUSIONS: Anti-inflammatory effects of, SHE against FD-induced keratinocyte inflammation is attributable to the suppression of upstream MAPK and NF-κB mediators. SHE indicated potential anti-inflammatory properties attenuating deteriorated skin barrier function in HaCaT keratinocytes. The effects are attributable to the polyphenols and other antioxidant compounds in SHE. Further studies could envisage the use of SHE for developing rejuvenating cosmetics.
Assuntos
Poeira , Inflamação/prevenção & controle , Queratinócitos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Sargassum/química , Sobrevivência Celular , Fracionamento Químico , Citocinas/genética , Citocinas/metabolismo , Etanol , Proteínas Filagrinas , Células HaCaT , Humanos , Inflamação/induzido quimicamente , Tamanho da Partícula , Extratos Vegetais/química , Espécies Reativas de OxigênioRESUMO
Sargassum horneri is an invasive brown seaweed that grows along the shallow coastal areas of the Korean peninsula, which are potentially harmful to fisheries and natural habitats in the areas where it is accumulated. Therefore, the author attempted to evaluate the anti-inflammatory mechanism of Sargachromenol isolated from S. horneri against particulate matter (PM)-stimulated RAW 264.7 macrophages. PM is a potent inducer of respiratory diseases such as lung dysfunctions and cancers. In the present study, the anti-inflammatory properties of Sargachromenol were validated using enzyme-linked immunosorbent assay (ELISA), Western blots, and RT-qPCR experiments. According to the results, Sargachromenol significantly downregulated the PM-induced proinflammatory cytokines, Prostaglandin E2 (PGE2), and Nitric Oxide (NO) secretion via blocking downstream activation of Toll-like receptor (TLR)-mediated nuclear factor kappa B (NF-κB) and MAPKs phosphorylation. Thus, Sargachromenol is a potential candidate for innovation in various fields including pharmaceuticals, cosmeceuticals, and functional food.
Assuntos
Anti-Inflamatórios/farmacologia , Benzopiranos/farmacologia , Extratos Vegetais/farmacologia , Sargassum , Animais , Anti-Inflamatórios/química , Organismos Aquáticos , Benzopiranos/química , Humanos , Macrófagos/metabolismo , Camundongos , Material Particulado , Extratos Vegetais/química , Células RAW 264.7/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Receptores Toll-Like/metabolismoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Sargassum horneri (Turner) C. Agardh (S. horneri), an edible brown marine algae, is known to have immunomodulatory effects and has been used in oriental medicine to treat inflammatory diseases. It is well known that ambient particulate matter (PM) is closely related to increased respiratory diseases inducing lung inflammation. AIM: Considering the use of Sargassum horneri in traditional medicine to treat inflammatory diseases, we hypothesized and investigated the use of Sargassum horneri containing polyphenols against PM-induced inflammatory responses. MATERIALS AND METHODS: In this study, we evaluated the impact of PM (majority <2.5 µm in diameter) on deep bronchial penetration ability upon inhalation and a therapeutic approach to mitigate its harmful effects using an ethanol extract of Sargassum horneri, an edible brown algae, containing polyphenols on a type II alveolar epithelial cell line, MLE-12. RESULTS: PM triggered mRNA expression of toll-like receptors (TLRs) TLR2/4/7, and those TLRs were significantly attenuated by Sargassum horneri extract (SHE). SHE further attenuated the phosphorylation of mitogen-activated protein kinase (MAPK) p38, extracellular signal-regulated kinase 1/2 (Erk1/2), and c-Jun NH (2)-terminal kinase (JNK), which were also activated in PM-exposed cells. Altogether, SHE subdued the PM-induced mRNA expression of pro-inflammatory cytokines (interleukin (IL)-1ß, tumor necrosis factor (TNF)-α, IL-6) and lung epithelial cell derived-chemokines (IL-8, monocyte chemoattractant protein-1 (MCP-1), and chemokine (C-C motif) ligand 5 (CCL5)). SHE also suppressed the mRNA expression of PM-induced pro-allergic cytokines thymic stromal lymphopoietin (TSLP) and interleukin (IL)-33. Furthermore, we showed that SHE suppressed the MAPK-dependent signaling pathway by attenuating receptor-associated factor (TRAF) 6 activation of proteins MyD88 and TNF. CONCLUSION: Taking all the data together, we suggest that the anti-inflammatory potential of SHE on PM-exposed MLE-12 cells is mediated by the inhibition of PM-triggered downstream signaling along the TLR2/4/7-MyD88-TRAF6 axis of MAPK signaling.
Assuntos
Anti-Inflamatórios/farmacologia , Inflamação/tratamento farmacológico , Polifenóis/farmacologia , Sargassum/química , Animais , Anti-Inflamatórios/isolamento & purificação , Linhagem Celular , Inflamação/patologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Camundongos , Fator 88 de Diferenciação Mieloide/metabolismo , Material Particulado/toxicidade , Polifenóis/isolamento & purificação , Transdução de Sinais/efeitos dos fármacos , Receptores Toll-Like/metabolismoRESUMO
The present study investigated the protective effects of Sargassum horneri (S. horneri) ethanol extract (SHE) against atopic dermatitis (AD), known as an abnormal immune response in house dust mite (HDM)/2,4-dinitrochlorobenzene (DNCB)-stimulated NC/Nga mice. The oral administration of SHE attenuated the AD symptoms, including the skin dermatitis severity, transepidermal water loss (TEWL), and ear edema in HDM/DNCB-stimulated mice. Moreover, the histological analysis revealed that SHE improved epidermal hyperplasia and hyperkeratosis, and reduced the dermal infiltrations of mast cells and eosinophils. Moreover, SHE downregulated the expression levels of cytokines (interleukin (IL)-6, IL-10, and interferon (IFN)-γ) and chemokines (Regulated on Activation, Normal T Cell Expressed and Secreted (RANTES), Eotaxin, and Thymus and activation-regulated chemokine (TARC)) by decreasing the expression levels of atopic initiators (IL-25 and IL-33) in HDM/DNCB-stimulated skin. The oral administration of SHE decreased the spleen size, reducing expression levels of AD-related cytokines (IL-4, IL-5, IL-6, IL-10, IL-13, IFN-γ, and TARC) by regulating the expressions of Tbx21 (T-bet), GATA Binding Protein 3 (GATA-3), and Signal transducer and activator of transcription 3 (STAT3). Moreover, SHE significantly attenuated the serum immunoglobulin (Ig)G1 and IgG2a levels in HDM/DNCB-stimulated mice. Collectively, these results suggest that S. horneri could be an ingredient of functional food against abnormal immune response.
Assuntos
Dermatite Atópica/tratamento farmacológico , Dermatite Atópica/imunologia , Dinitroclorobenzeno/imunologia , Alimento Funcional , Extratos Vegetais/administração & dosagem , Pyroglyphidae/imunologia , Sargassum/química , Administração Oral , Animais , Quimiocinas/genética , Quimiocinas/metabolismo , Citocinas/genética , Citocinas/metabolismo , Dermatite Atópica/genética , Dermatite Atópica/patologia , Feminino , Proteínas da Membrana Plasmática de Transporte de GABA/metabolismo , Expressão Gênica/efeitos dos fármacos , Imunoglobulina G/metabolismo , Camundongos , Fator de Transcrição STAT3/metabolismo , Índice de Gravidade de DoençaRESUMO
Eckol, a precursor compound belonging to the dibenzo-1,4-dioxin class of phlorotannins, is a phloroglucinol derivative that exerts various activities. In the present study, we investigated the antiallergic effects of eckol isolated from the marine brown algae, Ecklonia cava using immunoglobulin E (IgE)/bovine serum albumin (BSA)-stimulated mouse bone marrow-derived cultured mast cells (BMCMC) and a mouse model of anaphylaxis. Eckol inhibited IgE/BSA-induced BMCMC degranulation by reducing ß-hexosaminidase release. A flow cytometric analysis revealed that eckol decreases FcεRI expression on cell surface and IgE binding to the FcεRI in BMCMC. Moreover, eckol suppressed the production of the cytokines, interleukin (IL)-4, IL-5, IL-6, and IL-13 and the chemokine, thymus activation-regulated chemokine (TARC) by downregulating, IκB-α degradation and NF-κB nuclear translocation. Furthermore, it attenuated the passive cutaneous anaphylactic reaction induced by IgE/BSA-stimulation in the ear of BALB/c mice. These results suggest that eckol is a potential therapeutic candidate for the prevention and treatment of allergic disorders.
Assuntos
Anafilaxia/tratamento farmacológico , Antialérgicos , Dioxinas/farmacologia , Dioxinas/uso terapêutico , Imunoglobulina E/imunologia , Mastócitos/imunologia , Anafilaxia Cutânea Passiva/efeitos dos fármacos , Anafilaxia Cutânea Passiva/imunologia , Phaeophyceae/química , Fitoterapia , Anafilaxia/imunologia , Animais , Células Cultivadas , Dioxinas/isolamento & purificação , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BLRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Sargassum horneri is a nutrient rich edible brown seaweed with numerous biological properties found in shallow coastal areas of Korean peninsula. S. horneri traditionally used as a medicinal ingredient to treat several disease conditions such as hyperlipidemia, hypertension, heart disease, and inflammatory diseases (furuncle). However, to utilize S. horneri as an active ingredient for functional foods and human health applications requires to conform the bioactive properties and underlying mechanisms of those activities. AIM OF THE STUDY: Here, we investigated anti-inflammatory mechanisms of commercial grade 70% ethanol extract separated from S. horneri (SHE) on inflammatory response in particulate matter (PM)-induced MH-S lung macrophages; where PM in breathable air one of the major health concern in Korea. MATERIALS AND METHODS: We compared the anti-inflammatory effects of SHE on the activity of toll-like receptors (TLR) activation, NF-κB, MAPKs, and pro-inflammatory cytokine secretion in MH-S lung macrophages exposed to PM as a lung inflammation model. RESULTS: According to the results, PM-stimulation, induced the levels of NO, PGE2, TNF-α, IL-1ß, IL-6, iNOS, and COX2 (Pâ¯<â¯0.05) in MH-S macrophages. In addition, phosphorylation levels of NF-κB and MAPKs were also increased with the PM stimulation through the upregulated expression of TLR. However, SHE treatment significantly repressed the secretions of inflammatory cytokines and reduced protein expression such as PGE2, TNF-α, IL-6, IL-1ß, NF-κB, and MAPKs from PM-activated macrophages. Specifically, SHE inhibited the upregulated mRNA expression levels of TLR2, TLR3, TLR4, and TLR7 in PM-induced MH-S cells; known biomarkers of downstream activation of NF-κB and MAPKs. CONCLUSION: These results suggested that SHE is a potential inhibitor of PM-induced inflammatory responses in lung macrophages. Thus, SHE could inhibit PM-induced chronic inflammation in lungs via blocking TLR/NF-κB/MAPKs signal transduction. Therefore, it was concluded that SHE may be a useful substance to develop as functional product to reduce inflammation against PM-induced inflammation.
Assuntos
Anti-Inflamatórios/farmacologia , Inflamação/tratamento farmacológico , Macrófagos/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Material Particulado/efeitos adversos , Sargassum/química , Receptores Toll-Like/metabolismo , Animais , Linhagem Celular , Citocinas/metabolismo , Inflamação/induzido quimicamente , Inflamação/metabolismo , Macrófagos/metabolismo , Camundongos , Extratos Vegetais/farmacologia , República da Coreia , Transdução de Sinais/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Sargassum horneri is an edible brown seaweed with potential anti-inflammatory properties. The present study was designed to evaluate the anti-inflammatory properties of S. horneri using an in vivo mouse asthma model following exposure to particulate matter (PM). 7-8 week old BALB/c mice (20-25 g) were randomly divided into seven groups (n = 4) as follows: 1: no treatment, 2: OVA (ovalbumin) + PM, 3: OVA + PM + SHE (S. horneri ethanol extract) 200 mg kg-1, 4: OVA + PM + SHE 400 mg kg-1, 5: OVA + PM + prednisone 5 mg kg-1, 6: OVA only, and 7: PM only. All mice (except healthy controls) were sensitized on the first day by intraperitoneal injection of 10 µg OVA and 2 mg Al(OH)3 in 200 µL of saline. Starting from day 15, mice (except groups 1 and 6) were exposed to sonicated PM (5 mg m-3, 30 min day-1) through a nebulizer daily for 7 consecutive days. Mice exposed to PM and OVA showed up-regulated expression of MAPKs and pro-inflammatory cytokine production in the lungs. Furthermore, PM-exposed lungs had significantly reduced expression of Nrf2 and HO-1 genes. However, oral administration of the SHE reduced the phosphorylation levels of MAPKs, iNOS and COX2 expression levels, and mRNA expression levels of pro-inflammatory cytokines. In addition, SHE treated group mice had up-regulated anti-oxidant gene expression levels in the lungs compared to group 2. These findings demonstrate that oral administration of the SHE re-establishes PM-induced inflammation and oxidative stress in the lungs. Taken together, the SHE has therapeutic potential in preventing PM-induced inflammation and oxidative stress.
Assuntos
Asma/prevenção & controle , Material Particulado/efeitos adversos , Substâncias Protetoras/administração & dosagem , Sargassum/química , Animais , Asma/etiologia , Asma/genética , Asma/imunologia , Citocinas , Feminino , Heme Oxigenase-1/genética , Heme Oxigenase-1/imunologia , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/imunologia , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/imunologia , Extratos VegetaisRESUMO
BACKGROUND: Among the different kinds of pollution, air pollution continues to increase globally. East Asia is considered to be significantly affected. As a result, the populations in these regions face serious health issues including respiratory disorders. This study investigated the impact of fine dust (FD) particles (CRM No. 28) on macrophage cells as a model for alveolar lung cells. METHODS: The research focused on inflammation and oxidative stress induced by FD and Sargassum horneri (Turner) C. Agardh ethanol extract (SHE) as a potential treatment. S. horneri is a type of brown algae that has demonstrated anti-inflammatory effects against RAW 264.7 macrophages in previous studies. MTT, Griess, ELISA, western blotting, and mRNA expression analyses using PCR techniques were used in this study. RESULTS: The optimum FD concentration was determined to be 125 µg mL- 1. FD particles stimulated inflammatory mediators production (iNOS, COX-2, and PGE2) and pro-inflammatory cytokines (IL-1ß, IL-6, and TNF-α), leading to NO production. These mediators were dose-dependently downregulated by treatment with SHE. IL-6 and TNF-α were identified as biomarkers for FD. SHE treatment induced HO-1 and Nrf2 activity in a dose-dependent manner under FD stimulation. This confirmed the cytoprotective effect against oxidative stress induced via FD. Furthermore, treatment of the cells with a p38 MAPK inhibitor (SB202190) induced FD-stimulated NO production. CONCLUSIONS: The results suggest that SHE increases macrophage cellular resistance to FD-induced inflammation and oxidative stress, probably via the p38 MAPK pathway and Nrf2/HO-1 expression.
Assuntos
Antioxidantes/farmacologia , Heme Oxigenase-1/metabolismo , Inflamação/prevenção & controle , Fator 2 Relacionado a NF-E2/metabolismo , Extratos Vegetais/farmacologia , Sargassum/química , Animais , Sobrevivência Celular/efeitos dos fármacos , Poeira , Inflamação/induzido quimicamente , Camundongos , Estresse Oxidativo/efeitos dos fármacos , Células RAW 264.7 , Transdução de Sinais/efeitos dos fármacosRESUMO
Oxidative stress plays a crucial role in the progression of alcoholic liver diseases and substances of antioxidant property are of special interest for therapeutic purposes. We investigated the hepatoprotective effect of leaf extracts of Sasa quelpaertensis, an edible bamboo mainly cultivated in Jeju Island, South Korea. We examined the cytotoxicity of different extracts (distilled water, 20-80% EtOH) of S. quelpaertensis on HepG2 cells and their hepatoprotective effect on HepG2 cells stimulated by ethanol (800â¯mM, 24â¯h). Furthermore, we measured reactive oxygen species (ROS) production, ethanol toxicity induced cell death, and the activity of antioxidant enzymes. In in vivo experiments, liver damage was induced by oral administration of 5â¯g/kg ethanol with or without potent ethanol extract of S. quelpaertensis (10 or 100â¯mg/kg) 12â¯h interval for a total of 3 doses. Only 80% ethanol extract of S. quelpaertensis (SQEE80) exhibited cytoprotective effect on HepG2 cells against alcohol-induced toxicity. SQEE80 treatment (250, 500⯵g/mL) in ethanol exposed HepG2 cells showed significant attenuation of ROS production and ethanol toxicity induced cell death. Furthermore, SQEE80 markedly increased the activity of antioxidant enzyme glutathione peroxidase 1 in ethanol exposed HepG2 cells compared to ethanol stimulated cells. In in vivo experiments, SQEE80 treatment evidently suppressed the alcohol-induced histopathological changes in liver, serum ethanol content, and expression of cytochrome P450 2E1. Furthermore, SQEE80 significantly reversed the reduction of glutathione level in the ethanol challenged liver. Taken together, we suggest the possibility of developing SQEE80 as a natural hepatoprotective substance in attenuating alcohol-induced oxidative stress.
Assuntos
Antioxidantes/química , Hepatopatias Alcoólicas/tratamento farmacológico , Extratos Vegetais/química , Folhas de Planta/química , Sasa/química , Animais , Antioxidantes/uso terapêutico , Western Blotting , Sobrevivência Celular/efeitos dos fármacos , Células Hep G2/efeitos dos fármacos , Humanos , Imuno-Histoquímica , Camundongos , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/uso terapêuticoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Sasa quelpaertensis Nakai is an edible dwarf bamboo cultivated mainly in Jeju Island, South Korea and its leaf displays various health-promoting properties including antioxidant scavenging. AIM OF THE STUDY: In this study, we aimed at elucidating its hepatoprotective effect against alcohol-induced fatty liver. METHODS: In in vitro study, we evaluated the cytotoxicity and hepatoprotective effect of different solvent fractions (aqua, butanol, chloroform, ethyl acetate and hexane) of 80% EtOH extract of S. quelpaertensis Nakai leaf. In vivo experiment performed using binge alcohol consumption model. RESULTS: Although all five fractions (0-1000⯵g/mL) were non-cytotoxic to HepG2 cells, only ethyl acetate fraction (SQEA), rich in phenolic acids such as p-coumaric acid and flavonoids particularly myristin, showed hepatoprotective effect against EtOH (400â¯mM) in HepG2 cells. Furthermore, SQEA significantly decreased the ethanol induced cell death and enhanced the cell proliferation. In in vivo experiment using binge consumption model (5â¯g of EtOH/kg body weight in every 12â¯h for 3 times), SQEA treatment (10, 50 and 100â¯mg/kg) markedly reduced the alcohol induced histopathological changes and serum EtOH content, and reversed the reduction of glutathione level in ethanol challenged livers. Further, it suppressed the expression of cytochrome P450 2E1 (CYP2E1). In particular, SQEA activated AMP activated protein kinase (AMPK) pathway, and decreased the expression of tumor necrosis factor receptor-1 (TNFR1), which attenuated lipogenesis via decreased expression of fatty acid synthase (FAS). Inhibited lipogenesis due to SQEA treatment directed towards decreased perilipin-2 expression. These results indicate that SQEA has hypolipidemic effect which is mediated by decreased oxidative stress, increased fatty acid oxidation response and decreased lipogenesis. CONCLUSION: Our results suggest the possibility of developing SQEA as a natural hepatoprotective agent potent in attenuating alcohol-induced fatty liver.
Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Fígado Gorduroso Alcoólico/tratamento farmacológico , Flavonoides , Hidroxibenzoatos , Substâncias Protetoras , Sasa , Animais , Apoptose/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Citocromo P-450 CYP2E1/metabolismo , Fígado Gorduroso Alcoólico/metabolismo , Flavonoides/farmacologia , Flavonoides/uso terapêutico , Glutationa/metabolismo , Células Hep G2 , Humanos , Hidroxibenzoatos/farmacologia , Hidroxibenzoatos/uso terapêutico , Fígado/efeitos dos fármacos , Fígado/patologia , Camundongos Endogâmicos C57BL , Perilipina-2/metabolismo , Fitoterapia , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Folhas de Planta , Substâncias Protetoras/farmacologia , Substâncias Protetoras/uso terapêutico , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Triglicerídeos/metabolismoRESUMO
The leaves of Sasa quelpaertensis Nakai were extracted with 80% ethanol and further partitioned with n-hexane, chloroform, ethyl acetate, n-butanol, and aqueous fractions to evaluate the biological activity through assessment via various in vitro assays, including total phenol content; 1,1-diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azino-bis-(3-ethylbenzothiazothiazoline-6-sulfornic acid (ABTS) radical scavenging; reducing power; α-glucosidase and tyrosinase inhibitory; and alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) activity assays. The highest activity was found in the ethyl acetate fraction for all assays and showed stronger DPPH radical scavenging, reducing power, and tyrosinase inhibitory activity than the positive controls (butylated hydroxytoluene, α-tocopherol, and arbutin, respectively). When compared to the ethyl acetate fraction, the n-butanol fraction had lower rates, but it still demonstrated relatively high activity. The activity of the n-hexane fraction was high for DPPH and ABTS radical scavenging activity and contained significant amounts of phenol content, whereas the chloroform fraction possessed the highest reducing power, tyrosinase inhibitory, and ADH and ALDH activity, despite having the lowest phenol content when compared to the other fractions. These findings clearly indicate that S. quelpaertensis Nakai leaves can be a good natural source of antioxidants and tyrosinase inhibitors, as well as ADH and ALDH activity inducers, suggesting that may have potential for treating various diseases and improving human health.
Assuntos
Sasa , Disponibilidade Biológica , Extratos Vegetais , Folhas de Planta , SolventesRESUMO
Cells of the hematopoietic system are uniquely radiosensitive due to their rapid proliferation. Consequently, immune suppression readily and undesirably results from irradiation. Our previous studies demonstrated that geraniin isolated from Nymphaea tetragona var. angusta (water lily) had a protective effect on the splenocytes and intestinal tract of irradiated mice. This study was designed to assess the effectiveness of geraniin, an ellagitannin isolated from the water lily, in decreasing gamma ray irradiation-induced destruction of the hematopoietic system in mice. Geraniin treatment improved the survival time of bone marrow cells and maintained bone marrow integrity and also up-regulated the expression of stem cell receptors and the extent of cell mitosis. Geraniin also enhanced the proliferation and differentiation of immune cells that had been suppressed by irradiation. These results suggest geraniin is a promising agent for reconstituting hematopoietic cells after exposure to irradiation.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/efeitos da radiação , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/efeitos da radiação , Glucosídeos/farmacologia , Células-Tronco Hematopoéticas/citologia , Taninos Hidrolisáveis/farmacologia , Lesões Experimentais por Radiação/tratamento farmacológico , Protetores contra Radiação/farmacologia , Animais , Células da Medula Óssea , Células Cultivadas , Raios gama/efeitos adversos , Glucosídeos/administração & dosagem , Glucosídeos/isolamento & purificação , Glucosídeos/uso terapêutico , Células-Tronco Hematopoéticas/efeitos da radiação , Taninos Hidrolisáveis/administração & dosagem , Taninos Hidrolisáveis/isolamento & purificação , Taninos Hidrolisáveis/uso terapêutico , Injeções Intraperitoneais , Camundongos Endogâmicos C57BL , Nymphaea/química , Fitoterapia , Lesões Experimentais por Radiação/patologia , Protetores contra Radiação/uso terapêutico , Baço/citologiaRESUMO
CONTEXT: Beetroot [Beta vulgaris Linné (Chenopodiaceae)], a vegetable usually consumed as a food or a medicinal plant in Europe, has been reported to have antioxidant and anti-inflammatory properties. Since the lymphohematopoietic system is the most sensitive tissue to ionizing radiation, protecting it from radiation damage is one of the best ways to decrease detrimental effects from radiation exposure. OBJECTIVE: In this study, we evaluated the radio-protective effects of beetroot in hematopoietic stem cells (HSCs) and progenitor cells. MATERIALS AND METHODS: Beetroot extract was administered at a dose of 400 mg/mouse per os (p.o.) three times into C57BL/6 mice and, at day 10 after γ-ray irradiation, diverse molecular presentations were measured and compared against non-irradiated and irradiated mice with PBS treatments. Survival of beetroot-fed and unfed irradiated animal was also compared. RESULTS: Beetroot not only stimulated cell proliferation, but also minimized DNA damage of splenocytes. Beetroot also repopulated S-phase cells and increased Ki-67 or c-Kit positive cells in bone marrow. Moreover, beetroot-treated mice showed notable boosting of differentiation of HSCs into burst-forming units-erythroid along with increased production of IL-3. Also, beetroot-treated mice displayed enhancement in the level of hematocrit and hemoglobin as well as the number of red blood cell in peripheral blood. Beetroot diet improved survival rate of lethally exposed mice with a dose reduction factor (DRF) of 1.1. DISCUSSION AND CONCLUSION: These results suggest that beetroot has the potency to preserve bone marrow integrity and stimulate the differentiation of HSCs against ionizing radiation.
Assuntos
Beta vulgaris/química , Medula Óssea/efeitos dos fármacos , Raios gama/efeitos adversos , Hematínicos/farmacologia , Hematopoese/efeitos dos fármacos , Células-Tronco Hematopoéticas/efeitos dos fármacos , Tolerância Imunológica/efeitos dos fármacos , Fatores Imunológicos/farmacologia , Protetores contra Radiação/farmacologia , Animais , Apoptose/efeitos dos fármacos , Apoptose/efeitos da radiação , Medula Óssea/imunologia , Medula Óssea/patologia , Medula Óssea/efeitos da radiação , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/efeitos da radiação , Células Cultivadas , Dano ao DNA/efeitos dos fármacos , Relação Dose-Resposta a Droga , Hematínicos/isolamento & purificação , Hematopoese/efeitos da radiação , Células-Tronco Hematopoéticas/imunologia , Células-Tronco Hematopoéticas/patologia , Células-Tronco Hematopoéticas/efeitos da radiação , Tolerância Imunológica/efeitos da radiação , Fatores Imunológicos/isolamento & purificação , Interleucina-3/metabolismo , Camundongos Endogâmicos C57BL , Fitoterapia , Raízes de Plantas , Plantas Medicinais , Protetores contra Radiação/isolamento & purificação , Baço/efeitos dos fármacos , Baço/imunologia , Baço/efeitos da radiação , Fatores de Tempo , Irradiação Corporal Total/efeitos adversosRESUMO
Bearing pathologic and clinical similarities to human multiple sclerosis (MS), experimental autoimmune encephalomyelitis (EAE) is used as a murine model to test potential therapeutic agents for MS. Recently, we reported the protective effects of an acidic polysaccharide of Panax ginseng (APG) in C57BL/6 strain-dependent EAE, a model of primary progressive MS. In this study, we extend our previous findings on the therapeutic capacity of APG in relapsing-remitting EAE (rr-EAE), the animal model to closely mimic recurrent inflammatory demyelination lesions of relapsing-remitting MS. Treatments with APG led to a significant reduction of clinical symptoms and the relapse rate of EAE than vehicle treatments. Consistent with this, histological examination revealed that APG markedly modulated the infiltration of CD4[Formula: see text] T cells and CD11b[Formula: see text] macrophages into the spinal cord and the APG-treated CNS was devoid of demyelination and axonal damages. In addition, APG decreased the proliferation of peripheral PLP-reactive T cells and the production of pro-inflammatory factors such as IFN-[Formula: see text], IL-17 and TNF-[Formula: see text]. The fact that APG can induce clinically beneficial effects to distinct types of EAE furthers our understanding on the basis of its immunosuppression in EAE and, possibly, in MS. Our results suggest that APG may serve as a new therapeutic agent for MS as well as other human autoimmune diseases, and warrants continued evaluation for its translation into therapeutic application.
Assuntos
Encefalomielite Autoimune Experimental/tratamento farmacológico , Panax/química , Fitoterapia , Polissacarídeos/farmacologia , Polissacarídeos/uso terapêutico , Animais , Antígeno CD11b , Linfócitos T CD4-Positivos/imunologia , Células Cultivadas , Doenças Desmielinizantes , Modelos Animais de Doenças , Feminino , Humanos , Macrófagos/imunologia , Camundongos Endogâmicos C57BL , Esclerose Múltipla/tratamento farmacológico , Polissacarídeos/isolamento & purificação , Recidiva , Medula Espinal/imunologia , Medula Espinal/patologiaRESUMO
Dangyuja (Citrus grandis Osbeck), a citrus cultivated in southern Korea, has been used in traditional medicine for its anti-inflammatory effect. In this study, we investigated the anti-inflammatory potential of extract of Citrus grandis Osbeck (ECGO). In in vitro assays, ECGO treatment of concanavalin A (10µg/ml, for 24h) stimulated splenocytes showed significant reduction in CD44/CD62L+ T cell population and a marked decrease in the production of inflammatory cytokines IL-2, IFN-γ and IL-4. Interestingly, in vivo assays of ECGO topical treatment (100µg/20µl/ear) significantly mitigated the TPA (4µg/20µl/ear) induced edema induction and Myeloperoxidase activity. Anti-inflammatory potential of ECGO were further evidenced through its potent decrease in expression of inducible nitric oxide, cyclooxygenase-2, IL-1ß and TNF-α and suppressed homing of CD3+ T cells and F4/80+ macrophages to site of inflammation. This study emphasizes the possibility of developing ECGO as an alternative natural topical agent to combat inflammatory diseases.
Assuntos
Citrus , Concanavalina A/toxicidade , Edema/tratamento farmacológico , Extratos Vegetais/uso terapêutico , Baço/efeitos dos fármacos , Acetato de Tetradecanoilforbol/toxicidade , Animais , Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Relação Dose-Resposta a Droga , Edema/induzido quimicamente , Edema/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Dermatopatias/induzido quimicamente , Dermatopatias/tratamento farmacológico , Dermatopatias/metabolismo , Baço/citologia , Baço/metabolismoRESUMO
CONTEXT: Hallabong [(Citrus unshiu × C. sinensis) X C. reticulata)] (Rutaceae) is a hybrid citrus cultivated in temperate regions of South Korea. Its fruit is well-known for pharmacological properties. OBJECTIVE: This study examined the anti-inflammatory effect of 80% ethanol extract of Hallabong (HE) on concanavalin A (Con A)-stimulated splenocytes and mouse oedema model induced by 12-O-tetradecanoylphorbal acetate (TPA). MATERIALS AND METHODS: Murine splenocytes treated with HE were stimulated with Con A (10 µg/mL, for 24 h) were evaluated for T-cell population and production of inflammatory cytokines IL-2, IL-4 and IFN-γ. Anti-inflammatory effect of topically applied HE (100 µg/20 µL) on TPA (4 µg/20 µL/ear)-induced ear oedema was investigated in mouse model. RESULTS: HE-treated Con A-stimulated murine splenocytes showed a marked decrease in CD44/CD62L+ memory T-cell population, an important marker for anti-inflammatory activity, and a significant inhibition in the production of IL-2 and IFN-γ. HE treatment had reduced the mouse skin oedema (47%) and myeloperoxidase (MPO) activity significantly (40%) in TPA-challenged tissues. More importantly, immunohistochemical localization revealed the suppressed (p < 0.05) expression of inducible nitric oxide (iNOS), cyclooxygenase-2 (COX2). HE decreased the infiltration of CD3+ T cells and F4/80+ macrophages to the site of inflammation and a topical application of HE significantly suppressed the expression of TNF-α (20.2%). DISCUSSION AND CONCLUSION: A topical application of HE can exert a potential anti-inflammatory effect and HE can be explored further as a putative alternative therapeutic agent for inflammatory oedema.
Assuntos
Anti-Inflamatórios/uso terapêutico , Citrus , Modelos Animais de Doenças , Edema/tratamento farmacológico , Extratos Vegetais/uso terapêutico , Baço/efeitos dos fármacos , Acetato de Tetradecanoilforbol/toxicidade , Animais , Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Células Cultivadas , Citrus sinensis , Relação Dose-Resposta a Droga , Orelha , Edema/induzido quimicamente , Edema/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologiaRESUMO
The protective and therapeutic mechanism of bee venom acupuncture (BVA) in neurodegenerative disorders is not clear. We investigated whether treatment with BVA (0.25 and 0.8 mg/kg) at the Zusanli (ST36) acupoints, located lateral from the anterior border of the tibia, has a beneficial effect in a myelin basic protein (MBP)(68-82)-induced acute experimental autoimmune encephalomyelitis (EAE) rat model. Pretreatment (every 3 days from 1 h before immunization) with BVA was more effective than posttreatment (daily after immunization) with BVA with respect to clinical signs (neurological impairment and loss of body weight) of acute EAE rats. Treatment with BVA at the ST36 acupoint in normal rats did not induce the clinical signs. Pretreatment with BVA suppressed demyelination, glial activation, expression of cytokines [interferon (IFN)-γ, IL-17, IL-17A, tumor necrosis factor-alpha (TNF-α), and IL-1ß], chemokines [RANTES, monocyte chemotactic protein-1 (MCP-1), and macrophage inflammatory protein (MIP)-1α], and inducible nitric oxide synthase (iNOS), and activation of p38 mitogen-activated protein kinase (MAPK) and nuclear factor (NF)-κB (p65 and phospho-IκBα) signaling pathways in the spinal cord of acute EAE rats. Pretreatment with BVA decreased the number of CD4(+), CD4(+)/IFN-γ(+), and CD4(+)/IL-17(+) T cells, but increased the number of CD4(+)/Foxp3(+) T cells in the spinal cord and lymph nodes of acute EAE rats. Treatment with BVA at six placebo acupoints (SP9, GB39, and four non-acupoints) did not have a positive effect in acute EAE rats. Interestingly, onset and posttreatment with BVA at the ST36 acupoint markedly attenuated neurological impairment in myelin oligodendrocyte glycoprotein (MOG)(35-55)-induced chronic EAE mice compared to treatment with BVA at six placebo acupoints. Our findings strongly suggest that treatment with BVA with ST36 acupoint could delay or attenuate the development and progression of EAE by upregulating regulatory T cells and suppressing T-helper (Th) 17 and Th1 responses. These results warrant further investigation of BVA as a treatment for autoimmune disorders of the central nervous system.