RESUMO
Atopic dermatitis (AD) is a common chronic inflammatory skin disorder afflicting from infancy to adults with itching, scratching, and lichenification. We aimed to investigate the effects of esculetin from Fraxinus rhynchophylla on atopic skin inflammation. For induction of atopic skin inflammation, we exposed the ears of female BALB/c mice to house dust mite (Dermatophagoides farinae extract, DFE) and 2,4-dinitrochlorobenzene (DNCB) for 4â¯weeks. Oral administration of esculetin reduced the symptoms of DFE/DNCB-induced atopic skin inflammation, which were evaluated based on ear swelling and number of scratch bouts. The immunoglobulin (Ig) E, IgG2a, and histamine levels in serum were decreased and inflammatory cell infiltration in skin tissue was reduced by the esculetin. It suppressed production of Th1, Th2 and Th17-related cytokines such as tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-4, IL-13, IL-31 and IL-17 in the ear tissue. Furthermore, we investigated the effects of esculetin on activated keratinocytes, which are representative cells used for studying the pathogenesis of acute and chronic atopic skin inflammation. As results, esculetin suppressed gene expression of Th1, Th2 and Th17 cytokines and the activation of nuclear factor-κB and signal transducer and activator of transcription 1 in TNF-α/IFN-γ-stimulated keratinocytes. Taken together, these results imply that esculetin attenuated atopic skin inflammation, suggesting that esculetin could be a potential therapeutic candidate for the treatment of AD.
Assuntos
Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Citocinas/antagonistas & inibidores , Dermatite Alérgica de Contato/tratamento farmacológico , Umbeliferonas/farmacologia , Umbeliferonas/uso terapêutico , Animais , Antígenos de Dermatophagoides , Linhagem Celular , Citocinas/genética , Citocinas/imunologia , Dermatite Alérgica de Contato/imunologia , Dermatite Alérgica de Contato/metabolismo , Dinitroclorobenzeno , Feminino , Fraxinus , Humanos , Queratinócitos/efeitos dos fármacos , Queratinócitos/metabolismo , Camundongos Endogâmicos BALB C , NF-kappa B/metabolismo , Fator de Transcrição STAT1/metabolismo , Pele/efeitos dos fármacos , Pele/imunologiaRESUMO
Atopic dermatitis (AD) is a chronic relapsing inflammatory skin disorder. The present study investigated the effects of Amomum xanthioides extract (AXE) on ADlike skin inflammation using a Dermatophagoides farinae extract (DFE) and 2,4dinitrochlorobenzene (DNCB)induced mouse AD model. Hematoxylin and eosin staining results demonstrated that repeated DFE/DNCB exposure markedly increased the thickening of the dermis and epidermis, in addition to the infiltration of eosinophils and mast cells. However, oral administration of AXE reduced these histopathological alterations in a dosedependent manner. Elevated serum histamine, total and DFEspecific immunoglobulin E (IgE), and IgG2a were also decreased by treatment with AXE. In addition, reverse transcriptionquantitative polymerase chain reaction (RTqPCR) results demonstrated that the mRNA expression of tumor necrosis factor (TNF)α, interferon (IFN)γ, interleukin (IL)4, IL13, IL31 and IL17A was reduced in ear skin following AXE administration in AD mice. Fluorescenceactivated cell sorting demonstrated that the population of CD4+/IL4+, CD4+/IFNγ+ and CD4+/IL17A+ cells in draining lymph nodes was also significantly decreased in AXEtreated mice compared with AD mice without AXE treatment. Furthermore, keratinocytes that were stimulated with TNFα and IFNγ exhibited increased gene expression of proinflammatory cytokines and chemokines, including TNFα, IL1ß, IL6, IL8, CC motif chemokine ligand (CCL)17 and CCL22, as determined by RTqPCR. However, upregulation of these genes was reduced by AXE pretreatment. Based on these results, we hypothesize that AXE may be useful in the treatment of allergic skin inflammation, particularly AD.
Assuntos
Amomum/química , Anti-Inflamatórios/farmacologia , Dermatite Atópica/imunologia , Extratos Vegetais/farmacologia , Animais , Linhagem Celular , Dermatite Atópica/tratamento farmacológico , Dermatite Atópica/patologia , Modelos Animais de Doenças , Histamina/sangue , Histamina/imunologia , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Queratinócitos/efeitos dos fármacos , Queratinócitos/metabolismo , Camundongos , Pele/metabolismo , Pele/patologiaRESUMO
Phospho-cofilin (p-cofilin), which has a phosphate group on Ser-3, is involved in actin polymerization. Its dephosphorylated form promotes filopodia formation and cell migration by enhancing actin depolymerization. Protein phosphatase slingshot homologs (SSHs), known as dual-specificity phosphatases, catalyze hydrolytic removal of the Ser-3 phosphate group from phospho-cofilin. Aberrant SSH activity results in cancer metastasis, implicating SSHs as potential therapeutic targets for cancer metastasis. In this study, we screened 658 natural products purified from traditional oriental medicinal plants to identify three potent SSH inhibitors with submicromolar or single-digit micromolar Ki values: gossypol, hypericin, and sennoside A. The three compounds were purified from cottonseed, Saint John's wort, and rhubarb, respectively. Sennoside A markedly increased cofilin phosphorylation in pancreatic cancer cells, leading to impaired actin dynamics in pancreatic cancer cells with or without EGF stimulation and reduced motility and invasiveness in vitro and in vivo. Collaboratively, these results demonstrate that sennoside A is a novel inhibitor of SSHs and suggest that it may be valuable in the development of pharmaceutical drugs for treating cancer metastasis.
Assuntos
Antineoplásicos/farmacologia , Movimento Celular/efeitos dos fármacos , Invasividade Neoplásica/prevenção & controle , Neoplasias Pancreáticas/tratamento farmacológico , Fosfoproteínas Fosfatases/antagonistas & inibidores , Fosfoproteínas Fosfatases/metabolismo , Extrato de Senna/farmacologia , Fatores de Despolimerização de Actina/metabolismo , Actinas/metabolismo , Linhagem Celular Tumoral , Células Endoteliais da Veia Umbilical Humana , Humanos , Invasividade Neoplásica/patologia , Pâncreas/efeitos dos fármacos , Pâncreas/metabolismo , Pâncreas/patologia , Neoplasias Pancreáticas/metabolismo , Neoplasias Pancreáticas/patologia , Fosforilação/efeitos dos fármacos , SenosídeosRESUMO
The stress produced by the coupling of reactive oxygen species (ROS) and endoplasmic reticulum (ER) has been explored extensively, but little is known regarding their roles in the early development of mammalian embryos. Here, we demonstrated that the early development of in vitro-produced (IVP) bovine embryos was governed by the cooperative action between ROS and ER stress. Compared with the tension produced by 5% O2, 20% O2 significantly decreased the blastocyst formation rate and cell survival, which was accompanied by increases in ROS and in levels of sXBP-1 transcript, which is an ER stress indicator. In addition, treatment with glutathione (GSH), a ROS scavenger, decreased ROS levels, which resulted in increased blastocyst formation and cell survival rates. Importantly, levels of sXBP-1 and ER stress-associated transcripts were reduced by GSH treatment in developing bovine embryos. Consistent with this observation, tauroursodeoxycholate (TUDCA), an ER stress inhibitor, improved blastocyst developmental rate, trophectoderm proportion, and cell survival. Moreover, ROS and sXBP-1 transcript levels were markedly decreased by supplementation with TUDCA, suggesting a possible mechanism governing the mutual regulation between ROS and ER stress. Interestingly, knockdown of XBP-1 transcripts resulted in both elevation of ROS and decrease of antioxidant transcripts, which ultimately reduced in vitro developmental competence of bovine embryos. Based on these results, in vitro developmental competence of IVP bovine embryos was highly dependent on the coupled response between oxidative and ER stresses. These results increase our understanding of the mechanism(s) governing early embryonic development and may improve strategies for the generation of IVP embryos with high developmental competence.