RESUMO
Durian, known as the king of fruits, is rich in nutrients and bioactive phytochemicals. Propacin is a bioactive coumarinolignoid isolated from durian. In this study, we demonstrated its anti-inflammatory effect on lipopolysaccharide (LPS)-induced RAW264.7 cells and explored the underlying mechanism. Propacin suppressed nitric oxide (NO) and prostaglandin E2 (PGE2) production in LPS-stimulated macrophages significantly by downregulating the mRNA and protein expression of nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Moreover, propacin decreased the overexpression of the LPS-induced reactive oxygen species (ROS) and maintained the mitochondrial integrity in active macrophages. Furthermore, propacin inhibited the translocation of the nuclear factor-κB (NF-κB) p65 subunit into the nucleus and the phosphorylation of mitogen-activated protein kinases (MAPKs), especially JNK and ERK. Collectively, these data indicated that propacin may have the potential to be developed as a novel therapeutic agent for inflammatory-related diseases.
Assuntos
Anti-Inflamatórios/farmacologia , Bombacaceae/química , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Proteínas Quinases Ativadas por Mitógeno/imunologia , NF-kappa B/imunologia , Extratos Vegetais/farmacologia , Tianfenicol/farmacologia , Animais , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/imunologia , Lipopolissacarídeos/efeitos adversos , Camundongos , Proteínas Quinases Ativadas por Mitógeno/genética , NF-kappa B/genética , Óxido Nítrico Sintase/genética , Óxido Nítrico Sintase/imunologiaRESUMO
Xanthone is a phenolic compound found in a few higher plant families; it has a variety of biological activities, including antioxidant, anti-inflammatory, and anticancer properties. However, the molecular and cellular mechanisms underlying the activity of xanthone in allergic contact dermatitis (ACD) remain to be explored. Therefore, this study aimed to investigate the regulatory effects of xanthone in ACD in human keratinocytes (HaCaT cell), and human mast cell line (HMC-1 cell) in vitro and in an experimental murine model. The results demonstrated that treatment with xanthone reduced the production of pro-inflammatory cytokines and chemokines including interleukin (IL)-1ß, IL-6, IL-8, and expression of chemokines thymus and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC) in tumor necrosis factor (TNF)-α and interferon (IFN)-γ-stimulated HaCaT cells. Xanthone also suppressed the production of pro-inflammatory cytokines, chemokines, and allergic mediators in phorbol myristate acetate/A23187 calcium ionophore (PMACI)-stimulated HMC-1 cells. Xanthone significantly suppressed the phosphorylation of mitogen-activated protein kinases (MAPKs) and nuclear factor-kappa B (NF-κB) and activation of caspase-1 signaling pathway in vitro model. Additionally, xanthone administration alleviated 2,4-dinitrofluorobenzene (DNFB)-induced atopic dermatitis like-skin lesion by reducing the serum levels of immunoglobulin E (IgE), histamine, and pro-inflammatory cytokines and suppressing MAPKs phosphorylation. Xanthone administration also inhibited mortality due to compound 48/80-induced anaphylactic shock and suppressed the passive cutaneous anaphylaxis (PCA) reaction mediated by IgE. Collectively, these results suggest that xanthone has a potential for use in the treatment of allergic inflammatory diseases.
Assuntos
Anafilaxia/tratamento farmacológico , Antialérgicos/farmacologia , Dermatite Alérgica de Contato/tratamento farmacológico , Pele/efeitos dos fármacos , Xantonas/farmacologia , Administração Oral , Anafilaxia/induzido quimicamente , Anafilaxia/imunologia , Animais , Antialérgicos/uso terapêutico , Calcimicina/administração & dosagem , Calcimicina/imunologia , Linhagem Celular , Dermatite Alérgica de Contato/imunologia , Dermatite Alérgica de Contato/patologia , Dinitrofluorbenzeno/administração & dosagem , Dinitrofluorbenzeno/imunologia , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Humanos , Mediadores da Inflamação/metabolismo , Queratinócitos/efeitos dos fármacos , Queratinócitos/imunologia , Queratinócitos/patologia , Masculino , Mastócitos/efeitos dos fármacos , Mastócitos/imunologia , Mastócitos/patologia , Camundongos , Proteínas Quinases Ativadas por Mitógeno/imunologia , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fosforilação/efeitos dos fármacos , Fosforilação/imunologia , Pele/imunologia , Pele/patologia , Acetato de Tetradecanoilforbol/administração & dosagem , Acetato de Tetradecanoilforbol/imunologia , Xantonas/uso terapêutico , p-Metoxi-N-metilfenetilamina/imunologia , p-Metoxi-N-metilfenetilamina/toxicidadeRESUMO
BACKGROUND: Vaccinium oldhamii (V. oldhamii) has been reported to exert a variety of the pharmacological properties such as anti-oxidant activity, anti-cancer activity, and inhibitory activity of α-amylase and acetylcholinesterase. However, the anti-inflammatory activity of V. oldhamii has not been studied. In this study, we aimed to investigate anti-inflammatory activity of the stem extracts from V. oldhamii, and to elucidate the potential mechanisms in LPS-stimulated RAW264.7 cells. METHODS: Cell viability was evaluated by MTT assay. The determination of NO and PGE2 production was performed using Griess reagent and Prostaglandin E2 ELISA Kit, respectively. The change of mRNA or protein level was evaluated by RT-PCR and Western blot. RESULTS: Among VOS, VOL and VOF, the inhibitory effect of NO and PGE2 production induced by LPS was highest in VOS treatment. Thus, VOS was selected for the further study. VOS dose-dependently blocked LPS-induced NO and PGE2 production by inhibiting iNOS and COX-2 expression, respectively. VOS inhibited the expression of pro-inflammatory cytokines such as IL-1ß, IL-6 and TNF-α. In addition, VOS suppressed TRAP activity and attenuated the expression of the osteoclast-specific genes such as NFATc1, c-FOS, TRAP, MMP-9, cathepsin K, CA2, OSCAR and ATPv06d2. VOS inhibited LPS-induced NF-κB signaling activation through blocking IκB-α degradation and p65 nuclear accumulation. VOS inhibited MAPK signaling activation by attenuating the phosphorylation of ERK1/2, p38 and JNK. Furthermore, VOS inhibited ATF2 phosphorylation and blocked ATF2 nuclear accumulation. CONCLUSIONS: These results indicate that VOS may exert anti-inflammatory activity by inhibiting NF-κB and MAPK/ATF2 signaling. From these findings, VOS has potential to be a candidate for the development of chemopreventive or therapeutic agents for the inflammatory diseases.
Assuntos
Fator 2 Ativador da Transcrição/imunologia , Anti-Inflamatórios/farmacologia , Inflamação/imunologia , Macrófagos/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/imunologia , NF-kappa B/imunologia , Vaccinium/química , Fator 2 Ativador da Transcrição/genética , Animais , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/imunologia , Dinoprostona/imunologia , Humanos , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Inflamação/genética , Lipopolissacarídeos/efeitos adversos , Macrófagos/imunologia , Camundongos , Proteínas Quinases Ativadas por Mitógeno/genética , NF-kappa B/genética , Caules de Planta/química , Células RAW 264.7 , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Acute lung inflammation is one among the top of infectious diseases. It is a pulmonary dysfunctional disease. It breaks the physiological coordination in the structures and functions of respiratory system. There are a few effective treatments to minimize the mortality of acute lung inflammation. It was induced by Staphylococcus aureus (S. aureus) via nasal instillation of mice. The common ivy (Hedera helix) is the most significant medicinal plant and considered as a traditional medicinal plant. The most active ingredient in the extract of ivy plant was Hederacoside-C (HDC). The purpose of this study was to investigate its anti-inflammatory effects on induced acute lung inflammation in vivo and (RAW 264.7â¯cells) in vitro and to elucidate its anti-inflammatory mechanisms. HDC was administered intraperitoneally 1â¯h after infection until 24â¯h. The dose was repeated every 8â¯h for three successful doses. Mice treated with HDC significantly reduced the pulmonary edema, white blood cells, wet-dry ratio (W/D) and myeloperoxidase (MPO) activity. HDC attenuated protein expression levels of MAPKs including p38, ERK, JNK and NF-κB including p65 and IκB-α pathways analyzed by ELISA. HDC also suppressed the protein expressions of TLR2 & TLR4 detected by Western blot. HDC also downregulated the gene expression of pro-inflammatory cytokines including IL-6, IL-1ß and TNF-α, but upregulated the gene expression of an anti-inflammatory cytokine IL-10 analyzed by qRT-PCR. In conclusion, our results stated that HDC could inhibit the S. aureus induced acute lung inflammation and it may be a potential therapeutic drug against acute lung inflammation.
Assuntos
Anti-Inflamatórios/administração & dosagem , Medicamentos de Ervas Chinesas/administração & dosagem , Hedera/química , Ácido Oleanólico/análogos & derivados , Infecções Estafilocócicas/tratamento farmacológico , Infecções Estafilocócicas/imunologia , Staphylococcus aureus/imunologia , Receptor 2 Toll-Like/imunologia , Receptor 4 Toll-Like/imunologia , Animais , Feminino , Humanos , Interleucina-1beta/genética , Interleucina-1beta/imunologia , Interleucina-6/genética , Interleucina-6/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/imunologia , NF-kappa B/genética , NF-kappa B/imunologia , Ácido Oleanólico/administração & dosagem , Transdução de Sinais/efeitos dos fármacos , Infecções Estafilocócicas/genética , Staphylococcus aureus/genética , Staphylococcus aureus/fisiologia , Receptor 2 Toll-Like/genética , Receptor 4 Toll-Like/genética , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
The dried seeds of Cuminum cyminum L. have been traditionally used as food and medicine. To explore its chemical composition and anti-inflammatory activity, four new compounds (1-4) along with five known compounds (5-9) were isolated from the seeds in the present study. The chemical structures of the new compounds were identified as follows: methyl 3-((7H-purin-2-yl) amino)-3-(4-isopropylphenyl) propanoate (1), 8-(amino(4-isopropylphenyl)methyl)-5-hydroxy-2-(4-hydroxyphenyl)-7-methoxy-4-oxo-4H-chromene-6-carboxylic acid (2), (3,4,5-trihydroxy-6-((4-isopropylbenzyl)oxy)tetrahydro-2H-pyran-2-yl)methyl (E)-3-(4-propoxyphenyl)acrylate (3), and (3,4,5-trihydroxy-6-((5-hydroxy-2-(4-hydroxyphenyl)-4-oxo-4H-chromen-7-yl)oxy)tetrahydro-2H-pyran-2-yl)methyl 3-(4-isopropylphenyl)-2-methoxypropanoate (4). Compound 2, an atypical nitrogen-containing flavonoid, exhibited the most active inhibitory effect on nitride oxide, with IC50 of 5.25 µM in the lipopolysaccharide-stimulated RAW264.7 cell assay. Compound 2 was found to suppress the expression levels of inducible nitric oxide synthase and cyclooxygenase-2. Furthermore, it was revealed that both nuclear factor κB and mitogen-activated protein kinase were involved in the anti-inflammatory process of compound 2.
Assuntos
Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Cuminum/química , Flavonoides/química , Flavonoides/farmacologia , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Animais , Sobrevivência Celular/efeitos dos fármacos , Frutas/química , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Camundongos , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/imunologia , Estrutura Molecular , NF-kappa B/genética , NF-kappa B/imunologia , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/imunologia , Células RAW 264.7 , Sementes/químicaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Atopic dermatitis (AD), a disorder prevalent during childhood and adulthood, seriously affects the patient's quality of life. Although Huang-Lian-Jie-Du-Tang (HLJDT) has shown anti-inflammatory effects in previous studies, its effects and mechanism of action underlying AD disorder are still largely unknown. OBJECTIVE: This study explored the anti-inflammatory and immunomodulatory effects of HLJDT on the AD-like dermal disorder, induced in vitro by lipopolysaccharide (LPS)-triggered inflammation, and in vivo by 2,4-dinitrochlorobenzene (DNCB). MATERIALS AND METHODS: In vivo HLJDT effects were investigated by determining the severity of dermatitis, which consisted of observing signs of skin lesions, visually and through haematoxylin and eosin (HE) staining, in mouse ears and dorsal skin, measuring serum levels of interleukin (IL)-1α, IL-1ß, IL-2, IL-4, IL-5, IL-6, interferon (IFN)-γ, the tumour necrosis factor (TNF)-α, and determining the splenic index, number of splenic CD4+/CD8+ T-lymphocytes, as well as the phosphorylation levels of mitogen-activated protein kinases (including MAPKs-p38, ERK, and JNK), IκB-α, and nuclear factor kappa B (NF-κB) (p65) within dermal lesions. Morphological changes in LPS-induced inflammation were observed under a microscope, and ELISA and qPCR assays were used to measure IL-1α, IL-1ß, IL-6, and TNF-α expression levels. The protein expression levels of P-ERK/ERK, P-p38/p38, P-JNK/JNK, P-IKß-α, and P-p65 were measured through western blotting. Additionally, p65 expression was assessed by immunofluorescence, and LPS binding to RAW264.7â¯cell membrane was studied with laser confocal microscopy. RESULTS: HLJDT could remarkably mitigate DNCB-induced AD-like lesion symptoms, alleviating inflammatory mediator infiltration in mouse ears and dorsal skin tissue, down-regulating serum expression levels of IL-1α, IL-1ß, IL-2, IL-4, IL-5, IL-6, IFN-γ, and TNF-α, normalising the splenic CD4+/CD8+ T-lymphocyte ratio, and inactivating MAPKs (including p38, ERK, and JNK), IκB-α, and NF-κB (p65) in dorsal skin. Furthermore, HLJDT inhibited LPS-induced differentiation of RAW264.7â¯cells, as evidenced by the decreased protein and mRNA expression of IL-1α, IL-1ß, IL-6, and TNF-α. Additionally, it decreased ERK, p38, JNK, IKß-α, and p65 phosphorylation levels in the MAPKs/NF-κB pathway, inhibited p65 nuclear translocation, and reduced LPS binding to the RAW264.7â¯cell membrane. CONCLUSIONS: HLJDT significantly improved AD-like symptoms via inhibition of the MAPKs/NF-κB pathway. Therefore, administration of HLJDT might be a potential treatment for AD in the clinical setting.
Assuntos
Anti-Inflamatórios/uso terapêutico , Dermatite Atópica/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Fatores Imunológicos/uso terapêutico , Animais , Anti-Inflamatórios/farmacologia , Relação CD4-CD8 , Citocinas/imunologia , Dermatite Atópica/induzido quimicamente , Dermatite Atópica/imunologia , Dinitroclorobenzeno , Medicamentos de Ervas Chinesas/farmacologia , Fatores Imunológicos/farmacologia , Lipopolissacarídeos , Masculino , Camundongos , Proteínas Quinases Ativadas por Mitógeno/imunologia , NF-kappa B/imunologia , Células RAW 264.7 , Pele/efeitos dos fármacos , Pele/imunologiaRESUMO
A new acidic polysaccharide (GPTP-3) with a molecular weight of 2.49 × 106 Da was extracted and purified from Gynostemma pentaphyllum tea. Monosaccharide analysis revealed that GPTP-3 mainly comprised mannose (20.4%), glucuronic acid (17.4%), glucose (33.4%), and galactose (21.4%) (parentheses indicate the molar percentages). Immunostimulating assays indicated that GPTP-3 could markedly promote the secretion of NO, TNF-α, IL-1ß, and IL-6 in murine macrophage RAW264.7. TLR4 was found to be a recognized target of GPTP-3. Moreover, TLR4-related mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase (PI3K)/Akt, including ERK, JNK, p38, and Akt, were rapidly activated by GPTP-3 in RAW264.7 cells. Furthermore, GPTP-3 was found to induce the nuclear translocation of NF-κB subunit p65. All these findings suggest that MAPK, PI3K/Akt, and NF-κB pathways are involved in GPTP-3-induced macrophage activations, and GPTP-3 has the potential to be developed as a functional food with immunomodulatory functions.
Assuntos
Gynostemma/química , Fatores Imunológicos/farmacologia , Macrófagos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Polissacarídeos/farmacologia , Animais , Fatores Imunológicos/química , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Camundongos , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/imunologia , NF-kappa B/genética , NF-kappa B/imunologia , Extratos Vegetais/química , Polissacarídeos/química , Células RAW 264.7 , Chás de Ervas/análise , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
A novel polysaccharide (FCPW80-2) with a molecular weight of 1.21 × 105 Da was first isolated from Ficus carica through hot water extraction and several chromatographic methods. The structure of FCPW80-2 was determined by chemical and instrumental analysis. The results showed that the backbone of FCPW80-2 consists of (1â5)-linked α-l-Ara, (1â3,6)-linked ß-d-Man and (1â4,6)-linked ß-d-Gal. The branches of FCPW80-2 consist of (1â4)-linked α-d-Glc and (1â3)-linked ß-l-Rha terminated with (1â)-linked ß-d-Glc. In vitro immunomodulatory activity assays revealed that FCPW80-2 could markedly promote the secretion of cytotoxic molecules (NO) and cytokines (TNF-α and IL-6) as well as the phagocytosis of RAW264.7 macrophages. Moreover, TLR2 was found to be a pattern recognition receptor (PRR) of FCPW80-2, and its related mitogen-activated protein kinases (MAPKs), including p-ERK, p-JNK and p-p38, were rapidly upregulated by FCPW80-2 in RAW264.7 macrophages. Furthermore, FCPW80-2 could not only upregulate the expression of p-p65 and p-IκB-α, but also cause the translocation of nuclear factor-kappa B (NF-κB) p65 from cytosol to nuclei in RAW264.7 macrophages. The results demonstrated that MAPK and NF-κB signalling pathways participated in FCPW80-2-induced macrophage activation and FCPW80-2 could be developed as a potential immunomodulating functional food.
Assuntos
Ficus/química , Fatores Imunológicos/farmacologia , Extratos Vegetais/farmacologia , Polissacarídeos/farmacologia , Animais , Fatores Imunológicos/química , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Camundongos , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/imunologia , Inibidor de NF-kappaB alfa/genética , Inibidor de NF-kappaB alfa/imunologia , NF-kappa B/genética , NF-kappa B/imunologia , Extratos Vegetais/química , Polissacarídeos/química , Células RAW 264.7 , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Daidzein (DA) is an isoflavone that is primarily extracted from soy plants. This study evaluated the effects of supplementing laying broiler breeder hens with dietary DA on the immune function and growth performance of their offspring and the underlying mechanism. A total of 720 breeders were divided into three treatment groups that were fed either a control diet (CON), a DA-low-supplemented diet (DLS, CON+20 mg/kg DA), or a DA-high-supplemented diet (DHS, CON+100 mg/kg DA) for 8 weeks, and eggs were collected for hatching during the final week. The broiler offspring received a basal diet for 42 days, and blood, livers, and immune organs were collected at 21 and 42 days of age. DLS treatment promoted embryonic development and increased growth hormone levels, body weight, feed intake, and carcass traits on days 21 and 42 of broilers. Additionally, the IgA and IgG concentrations, antibody titers, and antioxidant capacity of broilers were increased at 21 days of age, and B lymphocyte differentiation was increased at 42 days. Besides, DLS treatment upregulated the expression of genes related to embryonic and muscle development in offspring and regulated mitogen-activated protein kinase (MAPK), transforming growth factor beta (TGF-ß), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), and Toll-like receptor signaling. DHS treatment decreased the percentage of abdominal fat in the broilers at 42 days, but it did not significantly affect embryonic development, growth performance, or IgA and IgG concentrations. In summary, providing dietary DA supplementation at 20 mg/kg to broiler breeders can improve their immune function and growth performance.
Assuntos
Galinhas/crescimento & desenvolvimento , Galinhas/imunologia , Suplementos Nutricionais/análise , Isoflavonas/administração & dosagem , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Galinhas/genética , Feminino , Imunoglobulina A/imunologia , Masculino , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/imunologia , NF-kappa B/genética , NF-kappa B/imunologia , TranscriptomaRESUMO
Chemotherapeutics are often used to inhibit the proliferation of cancer cells. However, they can also harm healthy cells and cause side effects such as immunosuppression. Especially traditional oriental medicines long used in Asia, may be beneficial candidates for the alleviation of immune diseases. Cervus nippon mantchuricus extract (NGE) is currently sold in the market as coffee and health drinks. However, NGE was not widely investigated and efficacy remain unclear and essentially nothing is known about their potential immune-regulatory properties. As a result, NGE induced the differentiation of RAW264.7 macrophage cells. NGE-stimulated RAW264.7 macrophage cells elevated cytokines levels and NO production. NGE-stimulated RAW264.7 macrophage cells activated MAPKs and NF-κB signaling pathways. NGE encouraged the immuno-enhancing effects in immunosuppressed short-term treated with NGE mice model. NGE or Red ginseng encouraged the immuno-enhancing effects in immunosuppressed long-term treated with NGE mice model. Our data clearly show that NGE contains immune-enhancing activity and can be used to treat immunodeficiency.
Assuntos
Osso e Ossos/imunologia , Cervos , Fatores Imunológicos/imunologia , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Extratos de Tecidos/imunologia , Animais , Western Blotting , Técnicas de Cultura de Células , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/imunologia , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/imunologia , Citocinas/efeitos dos fármacos , Citocinas/imunologia , Ensaio de Imunoadsorção Enzimática , Terapia de Imunossupressão , Masculino , Medicina Tradicional Coreana/métodos , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Quinases Ativadas por Mitógeno/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/imunologia , Modelos Animais , NF-kappa B/efeitos dos fármacos , NF-kappa B/imunologia , Óxido Nítrico/imunologia , Células RAW 264.7 , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologiaRESUMO
Tanshinones belong to a group of lipophilic constituents of Salvia miltiorrhiza Bunge (Danshen), which is widely used in traditional Chinese medicine. A deluge of studies demonstrated that tanshinones exert anti-inflammatory effects, but the underlying mechanisms remain unclear to date. This study investigated the anti-inflammatory effects and mechanisms of total tanshinones (TTN). TTN suppressed the expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) and the secretion of TNF-α, IL-6, and IL-1ß in RAW264.7 cells, bone marrow-derived macrophages, and THP-1 cells. TTN attenuated the LPS-induced transcriptional activity of NF-κB and decreased IκB-α and IKK phosphorylation and NF-κB/p65 nuclear translocation. Furthermore, TTN inhibited the LPS-induced transcriptional activity of AP-1, which was induced by the reduction of JNK1/2, ERK1/2, and p38MAPK phosphorylation. TTN blocked LPS-induced Toll-like receptor 4 (TLR4) dimerization, which consequently decreased MyD88 recruitment and TAK1 phosphorylation. In addition, TTN pretreatment effectively inhibited xylene-induced ear edema and LPS-induced septic death and improved LPS-induced acute kidney injury in mice. TTN exerts anti-inflammatory effects in vitro and in vivo by blocking TLR4 dimerization to activate MyD88-TAK1-NF-κB/MAPK signaling cascades, which provide the molecular basis of the anti-inflammatory effect of Danshen and suggest that TTN is a potential agent for the treatment of inflammatory diseases.
Assuntos
Abietanos/farmacologia , Anti-Inflamatórios/farmacologia , Edema/tratamento farmacológico , Fator 88 de Diferenciação Mieloide/imunologia , Salvia miltiorrhiza/química , Sepse/tratamento farmacológico , Receptor 4 Toll-Like/imunologia , Abietanos/isolamento & purificação , Animais , Anti-Inflamatórios/isolamento & purificação , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/imunologia , Orelha , Edema/induzido quimicamente , Edema/genética , Edema/imunologia , Regulação da Expressão Gênica , Humanos , Interleucina-1beta/genética , Interleucina-1beta/imunologia , Interleucina-6/genética , Interleucina-6/imunologia , Lipopolissacarídeos/farmacologia , Camundongos , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/imunologia , Fator 88 de Diferenciação Mieloide/genética , Inibidor de NF-kappaB alfa/genética , Inibidor de NF-kappaB alfa/imunologia , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/imunologia , Multimerização Proteica , Células RAW 264.7 , Sepse/induzido quimicamente , Sepse/genética , Sepse/imunologia , Transdução de Sinais , Células THP-1 , Receptor 4 Toll-Like/antagonistas & inibidores , Receptor 4 Toll-Like/genética , Fator de Transcrição AP-1/genética , Fator de Transcrição AP-1/imunologia , Fator de Transcrição RelA/genética , Fator de Transcrição RelA/imunologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Stearidonic acid (SDA, 18:4n-3) is an omega-3 polyunsaturated fatty acid present in oils derived from plants of the Boraginaceae family. In this study, we determined the anti-inflammatory effects of SDA isolated from echium oil on lipopolysaccharide (LPS)-induced inflammatory responses in RAW 264.7 macrophages. SDA significantly downregulated the levels of the inducible nitric oxide synthase (iNOS) protein, thereby suppressing the production of nitric oxide (NO) in LPS-stimulated RAW 264.7 cells. In addition, SDA inhibited the nuclear translocation and promoter activity of nuclear factor κB (NFκB) and the phosphorylation of mitogen-activated protein kinases (MAPK) such as extracellular signal regulated kinase 1/2, c-jun N terminal kinase, and p38 in LPS-stimulated RAW 264.7 cells. Our results showed that SDA exerted anti-inflammatory effects by suppressing iNOS-mediated NO production via inactivation of NFκB and MAPK signaling pathways.
Assuntos
Anti-Inflamatórios/farmacologia , Ácidos Graxos Ômega-3/farmacologia , Macrófagos/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/imunologia , NF-kappa B/imunologia , Animais , Anti-Inflamatórios/química , Boraginaceae/química , Ácidos Graxos Ômega-3/química , Lipopolissacarídeos/imunologia , Macrófagos/imunologia , Camundongos , Óxido Nítrico Sintase Tipo II/imunologia , Óleos de Plantas/química , Células RAW 264.7 , Transdução de Sinais/efeitos dos fármacosRESUMO
Lonicera caerulea L. berry polyphenols (LCBP) are considered as major components for bioactivity. This study aimed to clarify the molecular mechanisms by monitoring inflammatory and antioxidant mediator actions in lipopolysaccharide (LPS)-induced mouse paw edema and macrophage cell model. LCBP significantly attenuated LPS-induced paw edema (3.0 ± 0.1 to 2.8 ± 0.1 mm, P < 0.05) and reduced (P < 0.05) serum levels of monocyte chemotactic protein-1 (MCP-1, 100.9 ± 2.3 to 58.3 ± 14.5 ng/mL), interleukin (IL)-10 (1596.1 ± 424.3 to 709.7 ± 65.7 pg/mL), macrophage inflammatory protein (MIP)-1α (1761.9 ± 208.3 to 1369.1 ± 56.4 pg/mL), IL-6 (1262.8 ± 71.7 to 499.0 ± 67.1 pg/mL), IL-4 (93.3 ± 25.7 to 50.7 ± 12.5 pg/mL), IL-12(p-70) (580.4 ± 132.0 to 315.2 ± 35.1 pg/mL), and tumor necrosis factor-α (TNF-α, 2045.5 ± 264.9 to 1270.7 ± 158.6 pg/mL). Cell signaling analysis revealed that LCBP inhibited transforming growth factor ß activated kinase-1 (TAK1)-mediated mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) pathways, and enhanced the expression of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and manganese-dependent superoxide dismutase (MnSOD) in earlier response. Moreover, cyanidin 3-glucoside (C3G) and (-)-epicatechin (EC), two major components of LCBP, directly bound to TAK1. These data demonstrated that LCBP might inhibit LPS-induced inflammation by modulating both inflammatory and antioxidant mediators.
Assuntos
Antioxidantes/administração & dosagem , Edema/tratamento farmacológico , Mediadores da Inflamação/administração & dosagem , Lonicera/química , Extratos Vegetais/administração & dosagem , Polifenóis/administração & dosagem , Animais , Antioxidantes/química , Antioxidantes/isolamento & purificação , Quimiocina CCL2/genética , Quimiocina CCL2/imunologia , Edema/genética , Edema/imunologia , Frutas/química , Humanos , Mediadores da Inflamação/isolamento & purificação , Interleucina-12/genética , Interleucina-12/imunologia , Interleucina-4/genética , Interleucina-4/imunologia , Interleucina-6/genética , Interleucina-6/imunologia , Lipopolissacarídeos/efeitos adversos , Lipopolissacarídeos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos ICR , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/imunologia , NF-kappa B/genética , NF-kappa B/imunologia , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Polifenóis/química , Polifenóis/isolamento & purificaçãoRESUMO
Sea buckthorn has long been used as a functional food to regulate cholesterol, relieve angina, and diminish inflammation. Flavonoids are one of the main active components in sea buckthorn. We investigated the effects of sea buckthorn flavonoid (SF) treatment on two pathways that mediate inflammation, the mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-κB) pathways, to explore the anti-inflammatory activity of SFs in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. The LPS-induced over-production of nitric oxide (NO) and prostaglandin E2 (PGE2) was inhibited by SFs through a mechanism related to the modulatory effects of the inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) genes. Additionally, SFs downregulated the production and mRNA expression of pro-inflammatory cytokines including tumor necrosis factor (TNF)-α, interleukin (IL)-6 and IL-1ß. Moreover, SFs inhibited the phosphorylation of the p38 and stress-activated protein kinase/jun amino-terminal kinase (SAPK/JNK) MAPK pathways, and they reduced the nuclear translocation of NF-κB to prevent its activation by blocking the phosphorylation and degradation of inhibitor protein of NF-κB α (IκB-α). Based on these findings, SFs may exert their inhibitory effects on inflammation by regulating the release of inflammatory mediators through the MAPK and NF-κB pathways. SFs highlight the potential benefits of using functional foods with anti-inflammatory actions to combat inflammatory diseases.
Assuntos
Anti-Inflamatórios/farmacologia , Flavonoides/farmacologia , Hippophae/química , Inflamação/tratamento farmacológico , Macrófagos/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/imunologia , NF-kappa B/imunologia , Extratos Vegetais/farmacologia , Animais , Anti-Inflamatórios/isolamento & purificação , Flavonoides/isolamento & purificação , Humanos , Inflamação/genética , Inflamação/imunologia , Lipopolissacarídeos/efeitos adversos , Lipopolissacarídeos/imunologia , Macrófagos/imunologia , Camundongos , Proteínas Quinases Ativadas por Mitógeno/genética , NF-kappa B/genética , Extratos Vegetais/isolamento & purificação , Células RAW 264.7RESUMO
Four flavonoids (epicatechin, rutin, diosmin and luteolin) and 11 phenolic acids (gallic acid, gentisic acid, p-hydroxybezoic acid, vanillic acid, caffeic acid, p-coumaric acid, ferulic acid, sinapic acid, syringic acid, p-anisic acid and rosmarinic acid) were determined in the ethanolic extract of M. calabura Linn. fruit gathered in Taiwan. The extract suppressed the lipopolysaccharide-stimulated expressions of inducible nitric oxide synthase and cyclooxygenase-2 as well as the productions of nitric oxide, prostaglandin E2 and pro-inflammatory cytokines [tumour necrosis factor-α, interleukin (IL)-1ß and IL-6] in RAW264.7 macrophages. The extract modulated the inflammatory processes through inactivation of nuclear factor-κB (NF-κB), mitogen-activated protein kinases (MAPKs) p38 and c-Jun NH2-terminal kinase 1/2 (JNK1/2), and Janus kinase 2 (JAK2)/signal transducers and activators of transcription 1/3 (STAT1/3). Moreover, the activation of nuclear factor erythroid-2-related factor 2 (Nrf2) followed by inducing the production of heme oxygenase-1 (HO-1) is also related to the anti-inflammatory effect of the extract.
Assuntos
Anti-Inflamatórios/farmacologia , Mediadores da Inflamação/imunologia , Macrófagos/efeitos dos fármacos , Magnoliopsida/química , Extratos Vegetais/farmacologia , Animais , Anti-Inflamatórios/isolamento & purificação , Sobrevivência Celular/efeitos dos fármacos , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/imunologia , Dinoprostona , Frutas/química , Heme Oxigenase-1/genética , Heme Oxigenase-1/imunologia , Interleucina-1beta , Interleucina-6/genética , Interleucina-6/imunologia , Lipopolissacarídeos/efeitos adversos , Macrófagos/citologia , Macrófagos/imunologia , Camundongos , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/imunologia , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/imunologia , NF-kappa B , Óxido Nítrico , Extratos Vegetais/isolamento & purificação , Células RAW 264.7 , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Proteínas Quinases p38 Ativadas por Mitógeno/imunologiaRESUMO
BACKGROUND: Cheongsangbangpung-tang (CBT) is a traditional herbal formula used in Eastern Asia to treat heat-related diseases and swellings in the skin. The present study was conducted to evaluate the anti-inflammatory effects of cheongsangbangpung-tang extract (CBTE) both in vitro and in vivo. METHODS: The in vitro effects of CBTE on the lipopolysaccharide (LPS)-induced production of inflammation-related proteins were examined in RAW 264.7 cells. The levels of nitric oxide (NO) were measured with the Griess reagent. Inflammatory cytokines and prostaglandin E2 (PGE2) were detected using the enzyme-linked immunosorbent assay (ELISA) method. Inflammation-related proteins were detected by Western blot. The effect of CBTE on acute inflammation in vivo was evaluated using carrageenan (CA)-induced paw oedema. To evaluate the anti-inflammatory effect, paw oedema volume, thickness of the dorsum and ventrum pedis skin, number of infiltrated inflammatory cells, and number of COX-2-, iNOS-immunoreactive cells were measured. RESULTS: In an in vitro study, CBTE inhibited the production of NO and PGE2 and also decreased the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) activity, interleukin (IL)-1ß, IL-6 and tumuor necrosis factor-α. In LPS-activated macrophages, nuclear factor-kappaB (NF-κB) and mitogen-activated protein kinase (MAPK) signalling is a pivotal pathway in the inflammatory process. These plausible molecular mechanisms increased the phosphorylation of I-κBα, while the activation of NF-κB and the phosphorylation of MAPK by LPS were blocked by CBTE treatment. In our in vivo study, a CA-induced acute oedematous paw inflammation rat model was used to evaluate the anti-inflammatory effect of CBTE. CBTE significantly reduced the increases in paw swelling, skin thicknesses, infiltrated inflammatory cells and iNOS-, COX-2 positive cells induced by CA injection. CONCLUSIONS: Based on these results, CBTE should favourably inhibit the acute inflammatory response through modulation of NF-κB activation and MAPK phosphorylation. Furthermore, the inhibition of CBTE in rat paw oedema induced by CA is considered to be clear evidence that CBTE may be a useful source to treat inflammation.
Assuntos
Anti-Inflamatórios/administração & dosagem , Edema/tratamento farmacológico , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/imunologia , Extratos Vegetais/administração & dosagem , Animais , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/imunologia , Edema/genética , Edema/imunologia , Humanos , Interleucina-6/genética , Interleucina-6/imunologia , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Masculino , Camundongos , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/imunologia , NF-kappa B/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/imunologia , Fosforilação/efeitos dos fármacos , Células RAW 264.7 , Ratos , Ratos Sprague-DawleyRESUMO
The mitogen-activated protein kinases (MAPKs) family is known to mediate various biological processes in response to diverse environmental pollutants. Although MAPKs are well characterized and studied in vertebrates, in invertebrates the cross-reactivities of MAPKs antibodies were not clearly known in response to environmental pollutants due to limited information of antibody epitopes with material resources for invertebrates. In this paper, we performed phylogenetic analysis of MAPKs genes in the marine rotifer Brachionus koreanus and the copepods Paracyclopina nana and Tigriopus japonicus. Also in rotifer and copepods, several studies of Western blot of MAPK signaling pathways were shown in response to environmental pollutants, including multi-walled carbon nanotubes (MWCNTs), water-accommodated fractions (WAFs) of crude oil, and microplastics. This paper will provide a better understanding of the underlying mechanistic scenario in terms of cross-reactivities of mammalian antibodies in rotifer and copepod.
Assuntos
Copépodes/imunologia , Ecotoxicologia/métodos , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/imunologia , Rotíferos/imunologia , Animais , Anticorpos/imunologia , Western Blotting , Copépodes/genética , Reações Cruzadas , Regulação Enzimológica da Expressão Gênica , Mamíferos/imunologia , Nanotubos de Carbono/toxicidade , Petróleo/metabolismo , Filogenia , Plásticos/toxicidade , Rotíferos/genética , Poluentes Químicos da Água/toxicidadeRESUMO
Excessive microglial cell activation is related to the progression of chronic neuro-inflammatory disorders. Heme oxygenase-1 (HO-1) expression mediated by the NFE2-related factor (Nrf-2) pathway is a key regulator of neuro-inflammation. Nardostachys chinensis is used as an anti-malarial, anti-nociceptive, and neurotrophic treatment in traditional Asian medicines. In the present study, we examined the effects of an ethyl acetate extract of N. chinensis (EN) on the anti-neuro-inflammatory effects mediated by HO-1 up-regulation in Salmonella lipopolysaccharide (LPS)- or Staphylococcus aureus lipoteichoic acid (LTA)-stimulated BV2 microglial cells. Our results indicated that EN suppressed pro-inflammatory cytokine production and induced HO-1 transcription and translation through Nrf-2/antioxidant response element (ARE) signaling. EN markedly inhibited LPS- and LTA-induced activation of nuclear factor-kappa B (NF-κB) as well as phosphorylation of mitogen-activated protein kinases (MAPKs) and signal transducer and activator of transcription (STAT). Furthermore, EN protected hippocampal HT22 cells from indirect neuronal toxicity mediated by LPS- and LTA-treated microglial cells. These results suggested that EN impairs LPS- and LTA-induced neuro-inflammatory responses in microglial cells and confers protection against indirect neuronal damage to HT22 cells. In conclusion, our findings indicate that EN could be used as a natural anti-neuro-inflammatory and neuroprotective agent.
Assuntos
Anti-Inflamatórios/farmacologia , Microglia/efeitos dos fármacos , Nardostachys/química , Fármacos Neuroprotetores/farmacologia , Extratos Vegetais/farmacologia , Linhagem Celular , Heme Oxigenase-1/genética , Heme Oxigenase-1/imunologia , Humanos , Lipopolissacarídeos/efeitos adversos , Microglia/citologia , Microglia/imunologia , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/imunologia , NF-kappa B/genética , NF-kappa B/imunologia , Ácidos Teicoicos/efeitos adversosRESUMO
We herein report the immunostimulatory effect of spinach aqueous extract (SAE) on mouse macrophage-like J774.1 cells and mouse primary peritoneal macrophages. SAE significantly enhanced the production of interleukin (IL)-6 and tumor necrosis factor-α by both J774.1 cells and peritoneal macrophages by enhancing the expression levels of these cytokine genes. In addition, the phagocytosis activity of J774.1 cells was facilitated by SAE. Immunoblot analysis revealed that SAE activates mitogen-activated protein kinase and nuclear factor-κB cascades. It was found that SAE activates macrophages through not only TLR4, but also other receptors. The production of IL-6 was significantly enhanced by peritoneal macrophages from SAE-administered BALB/c mice, suggesting that SAE has a potential to stimulate macrophage activity in vivo. Taken together, these data indicate that SAE would be a beneficial functional food with immunostimulatory effects on macrophages.
Assuntos
Adjuvantes Imunológicos/farmacologia , Interleucina-6/agonistas , Macrófagos Peritoneais/efeitos dos fármacos , Extratos Vegetais/farmacologia , Spinacia oleracea/química , Fator de Necrose Tumoral alfa/agonistas , Animais , Linhagem Celular , Feminino , Expressão Gênica , Interleucina-6/genética , Interleucina-6/imunologia , Macrófagos Peritoneais/citologia , Macrófagos Peritoneais/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/imunologia , NF-kappa B/agonistas , NF-kappa B/genética , NF-kappa B/imunologia , Fagocitose/efeitos dos fármacos , Cultura Primária de Células , RNA Mensageiro/agonistas , RNA Mensageiro/genética , RNA Mensageiro/imunologia , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/imunologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Cajaninstilbene acid (CSA) is one of the active components isolated from pigeon pea leaves. In this study, anti-inflammatory effects of CSA and its synthesized derivatives were fully valued with regard to their activities on the production of nitric oxide (NO) and pro-inflammatory cytokines tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in vitro cell model, as well as their impacts on the migration of neutrophils and macrophages in fluorescent protein labeled zebrafish larvae model by live image analysis. Furthermore, the anti-inflammatory mechanism of this type of compounds was clarified by western-blot and reverse transcription-polymerase chain reaction (RT-PCR). The results showed that CSA, as well as its synthesized derivatives 5c, 5e and 5h, exhibited strong inhibition activity on the release of NO and inflammatory factor TNF-α and IL-6 in lipopolysaccharides (LPS)-stimulated murine macrophages. CSA and 5c greatly inhibited the migration of neutrophils and macrophages in injury zebrafish larvae. CSA and 5c treatment greatly inhibited the phosphorylation of proteins involved in nuclear factor kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) pathways. Moreover, we found that peroxisome proliferator-activated receptor gamma (PPARγ) inhibitor GW9662 could reverse partly the roles of CSA and 5c, and CSA and 5c treatment greatly resist the decrease of PPARγ mRNA and protein induced by LPS stimulation. Our results identified the promising anti-inflammatory effects of CSA and its derivatives, which may serve as valuable anti-inflammatory lead compound. Additionally, the mechanism studies demonstrated that the anti-inflammatory activity of CSA and its derivative is associated with the inhibition of NF-κB and MAPK pathways, relying partly on resisting the LPS-induced decrease of PPARγ through improving its expression.