RESUMO
Qin Pi (Fraxinus chinensis Roxb.) is commercially used in healthcare products for the improvement of intestinal function and gouty arthritis in many countries. Three new secoiridoid glucosides, (8E)-4''-O-methylligstroside (1), (8E)-4''-O-methyldemethylligstroside (2), and 3'',4''-di-O-methyl-demethyloleuropein (3), have been isolated from the stem bark of Fraxinus chinensis, together with 23 known compounds (4-26). The structures of the new compounds were established by spectroscopic analyses (1D, 2D NMR, IR, UV, and HRESIMS). Among the isolated compounds, (8E)-4''-O-methylligstroside (1), (8E)-4''-O-methyldemethylligstroside (2), 3'',4''-di-O-methyldemethyloleuropein (3), oleuropein (6), aesculetin (9), isoscopoletin (11), aesculetin dimethyl ester (12), fraxetin (14), tyrosol (21), 4-hydroxyphenethyl acetate (22), and (+)-pinoresinol (24) exhibited inhibition (IC50 ≤ 7.65 µg/mL) of superoxide anion generation by human neutrophils in response to formyl-L-methionyl-L-leuckyl-L-phenylalanine/cytochalasin B (fMLP/CB). Compounds 1, 9, 11, 14, 21, and 22 inhibited fMLP/CB-induced elastase release with IC50 ≤ 3.23 µg/mL. In addition, compounds 2, 9, 11, 14, and 21 showed potent inhibition with IC50 values ≤ 27.11 µM, against lipopolysaccharide (LPS)-induced nitric oxide (NO) generation. The well-known proinflammatory cytokines, tumor necrosis factor-alpha (TNF-α) and interleukin 6 (IL-6), were also inhibited by compounds 1, 9, and 14. Compounds 1, 9, and 14 displayed an anti-inflammatory effect against NO, TNF-α, and IL-6 through the inhibition of activation of MAPKs and IκBα in LPS-activated macrophages. In addition, compounds 1, 9, and 14 stimulated anti-inflammatory M2 phenotype by elevating the expression of arginase 1 and Krüppel-like factor 4 (KLF4). The above results suggested that compounds 1, 9, and 14 could be considered as potential compounds for further development of NO production-targeted anti-inflammatory agents.
Assuntos
Anti-Inflamatórios/farmacologia , Fraxinus/química , Regulação da Expressão Gênica/efeitos dos fármacos , Glucosídeos Iridoides/farmacologia , Casca de Planta/química , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/classificação , Anti-Inflamatórios/isolamento & purificação , Citocalasina B/antagonistas & inibidores , Citocalasina B/farmacologia , Regulação da Expressão Gênica/imunologia , Humanos , Interleucina-6/genética , Interleucina-6/imunologia , Glucosídeos Iridoides/química , Glucosídeos Iridoides/classificação , Glucosídeos Iridoides/isolamento & purificação , Fator 4 Semelhante a Kruppel , Fatores de Transcrição Kruppel-Like/genética , Fatores de Transcrição Kruppel-Like/imunologia , Elastase de Leucócito/imunologia , Elastase de Leucócito/metabolismo , Lipopolissacarídeos/antagonistas & inibidores , Lipopolissacarídeos/farmacologia , MAP Quinase Quinase 4/genética , MAP Quinase Quinase 4/imunologia , Camundongos , Estrutura Molecular , N-Formilmetionina Leucil-Fenilalanina/antagonistas & inibidores , N-Formilmetionina Leucil-Fenilalanina/farmacologia , Inibidor de NF-kappaB alfa/genética , Inibidor de NF-kappaB alfa/imunologia , Neutrófilos/citologia , Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Óxido Nítrico/antagonistas & inibidores , Óxido Nítrico/metabolismo , Extratos Vegetais/química , Cultura Primária de Células , Células RAW 264.7 , Relação Estrutura-Atividade , Superóxidos/antagonistas & inibidores , Superóxidos/metabolismo , 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: Endothelial cell inflammation is a central event in the pathogenesis of numerous cardiovascular diseases, including sepsis and atherosclerosis. Triptolide, a principal bioactive ingredient of Traditional Chinese Medicine Tripterygium wilfordii Hook.F., displays anti-inflammatory actions in vivo. However, the mechanisms underlying these beneficial effects remain undetermined. The present study investigated the effects and possible mechanisms of triptolide on lipopolysaccharide (LPS)-induced inflammatory responses in human umbilical vein endothelial cells (HUVECs). METHODS: The effects of triptolide on the LPS-induced production and expression of inflammatory molecules, monocyte adhesion and activation of nuclear factor (NF)-κB pathway were examined in cultured HUVECs. RESULTS: In cultured HUVECs, pre-treatment with triptolide dose-dependently attenuated LPS-induced cytokine and chemokine production, adhesion molecule expression and monocyte adhesion. Mechanistically, triptolide was found to dose-dependently inhibit the LPS-induced increases in the DNA binding activity of NF-κB p65 associated with attenuating IκBα phosphorylation and its degradation. Additionally, the present study revealed that triptolide inhibited LPS-triggered NF-κB transcriptional activation in a dose-dependent manner. CONCLUSIONS: The results of the present study indicated that triptolide suppresses the inflammatory response of endothelial cells possibly via inhibition of NF-κB activation.
Assuntos
Anti-Inflamatórios/farmacologia , Diterpenos/farmacologia , Medicamentos de Ervas Chinesas/farmacologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , NF-kappa B/imunologia , Fenantrenos/farmacologia , Tripterygium/química , Compostos de Epóxi/farmacologia , Células Endoteliais da Veia Umbilical Humana/imunologia , Humanos , Lipopolissacarídeos/efeitos adversos , Monócitos/efeitos dos fármacos , Monócitos/imunologia , Inibidor de NF-kappaB alfa/genética , Inibidor de NF-kappaB alfa/imunologia , NF-kappa B/genética , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Self-assembling natural drug hydrogels formed without structural modification and able to act as carriers are of interest for biomedical applications. A lack of knowledge about natural drug gels limits there current application. Here, we report on rhein, a herbal natural product, which is directly self-assembled into hydrogels through noncovalent interactions. This hydrogel shows excellent stability, sustained release and reversible stimuli-responses. The hydrogel consists of a three-dimensional nanofiber network that prevents premature degradation. Moreover, it easily enters cells and binds to toll-like receptor 4. This enables rhein hydrogels to significantly dephosphorylate IκBα, inhibiting the nuclear translocation of p65 at the NFκB signalling pathway in lipopolysaccharide-induced BV2 microglia. Subsequently, rhein hydrogels alleviate neuroinflammation with a long-lasting effect and little cytotoxicity compared to the equivalent free-drug in vitro. This study highlights a direct self-assembly hydrogel from natural small molecule as a promising neuroinflammatory therapy.
Assuntos
Antraquinonas/administração & dosagem , Anti-Inflamatórios/administração & dosagem , Portadores de Fármacos/química , Inflamação/tratamento farmacológico , Microglia/efeitos dos fármacos , Animais , Antraquinonas/química , Antraquinonas/farmacocinética , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacocinética , Linhagem Celular , Preparações de Ação Retardada/administração & dosagem , Preparações de Ação Retardada/química , Preparações de Ação Retardada/farmacocinética , Portadores de Fármacos/administração & dosagem , Portadores de Fármacos/farmacocinética , Avaliação Pré-Clínica de Medicamentos , Liberação Controlada de Fármacos , Hidrogéis/administração & dosagem , Hidrogéis/química , Hidrogéis/farmacocinética , Inflamação/imunologia , Inflamação/patologia , Lipopolissacarídeos/imunologia , Camundongos , Microglia/imunologia , Microglia/patologia , Microscopia Eletrônica de Varredura , Inibidor de NF-kappaB alfa/imunologia , Inibidor de NF-kappaB alfa/metabolismo , Nanofibras/administração & dosagem , Nanofibras/química , Nanofibras/ultraestrutura , Fosforilação/efeitos dos fármacos , Fosforilação/imunologia , Compostos Fitoquímicos/administração & dosagem , Compostos Fitoquímicos/química , Compostos Fitoquímicos/farmacocinética , Rheum/química , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologia , Receptor 4 Toll-Like/metabolismo , Fator de Transcrição RelA/imunologia , Fator de Transcrição RelA/metabolismoRESUMO
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
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.