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1.
Front Immunol ; 12: 624197, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33815376

RESUMO

Vaccines have played a pivotal role in improving public health, however, many infectious diseases lack an effective vaccine. Controlling the spread of infectious diseases requires continuing studies to develop new and improved vaccines. Our laboratory has been investigating the immune enhancing mechanisms of Toll-like receptor (TLR) ligand-based adjuvants, including the TLR2 ligand Neisseria meningitidis outer membrane protein, PorB. Adjuvant use of PorB increases costimulatory factors on antigen presenting cells (APC), increases antigen specific antibody production, and cytokine producing T cells. We have demonstrated that macrophage expression of MyD88 (required for TLR2 signaling) is an absolute requirement for the improved antibody response induced by PorB. Here-in, we specifically investigated the role of subcapsular CD169+ marginal zone macrophages in antibody production induced by the use of TLR-ligand based adjuvants (PorB and CpG) and non-TLR-ligand adjuvants (aluminum salts). CD169 knockout mice and mice treated with low dose clodronate treated animals (which only remove marginal zone macrophages), were used to investigate the role of these macrophages in adjuvant-dependent antibody production. In both sets of mice, total antigen specific immunoglobulins (IgGs) were diminished regardless of adjuvant used. However, the greatest reduction was seen with the use of TLR ligands as adjuvants. In addition, the effect of the absence of CD169+ macrophages on adjuvant induced antigen and antigen presenting cell trafficking to the lymph nodes was examined using immunofluorescence by determining the relative extent of antigen loading on dendritic cells (DCs) and antigen deposition on follicular dendritic cells (FDC). Interestingly, only vaccine preparations containing PorB had significant decreases in antigen deposition in lymphoid follicles and germinal centers in CD169 knockout mice or mice treated with low dose clodronate as compared to wildtype controls. Mice immunized with CpG containing preparations demonstrated decreased FDC networks in the mice treated with low dose clodronate. Conversely, alum containing preparations only demonstrated significant decreases in IgG in CD169 knockout mice. These studies stress that importance of subcapsular macrophages and their unique role in adjuvant-mediated antibody production, potentially due to an effect of these adjuvants on antigen trafficking to the lymph node and deposition on follicular dendritic cells.


Assuntos
Adjuvantes Imunológicos/farmacologia , Compostos de Alúmen/farmacologia , Imunogenicidade da Vacina , Macrófagos/efeitos dos fármacos , Oligodesoxirribonucleotídeos/farmacologia , Ovalbumina/farmacologia , Porinas/farmacologia , Lectina 1 Semelhante a Ig de Ligação ao Ácido Siálico/metabolismo , Receptores Toll-Like/agonistas , Animais , Ácido Clodrônico/farmacologia , Células Dendríticas Foliculares/efeitos dos fármacos , Células Dendríticas Foliculares/imunologia , Células Dendríticas Foliculares/metabolismo , Imunoglobulina G/sangue , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos Endogâmicos C57BL , Camundongos Knockout , Oligodesoxirribonucleotídeos/imunologia , Ovalbumina/imunologia , Porinas/imunologia , Lectina 1 Semelhante a Ig de Ligação ao Ácido Siálico/genética , Transdução de Sinais , Receptores Toll-Like/metabolismo , Vacinação
2.
Respir Physiol Neurobiol ; 288: 103642, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33609775

RESUMO

OBJECTIVE: To investigate the effects of serine protease inhibitor 3n (SerpinA3n) in a neonatal mouse model of asthma. METHODS: The study utilized a neonatal mouse ovalbumin (OVA) sensitization model of asthma. Wild type (WT) and SerpinA3n-/- mice were randomly divided into WT/SerpinA3n-/- + saline, WT/SerpinA3n-/- + OVA, WT/SerpinA3n-/- + OVA + rSerpinA3n (recombinant mouse SerpinA3n protein), and WT/SerpinA3n-/- + OVA + DEX (dexamethasone, positive control) groups followed by hematoxylin-eosin (HE) staining, Masson's trichrome stainings, Sircol soluble collagen assay, quantitative real time polymerase chain reaction (qRT-PCR), Western Blot and enzyme linked immunosorbent assay (ELISA). RESULTS: OVA-induced neonatal mice showed the increases in airway hyper-reactivity with the up-regulated total cells, eosinophil, lymphocyte and neutrophil in bronchoalveolar lavage fluid (BALF), which was much higher in WT + OVA + rSerpinA3n group (P < 0.05). SerpinA3n-/- suppressed the serum concentrations of total immunoglobulin E (IgE) and OVA-specific IgG1 in OVA-induced asthmatic mice, and alleviated the pathological changes of lung tissues, which was reversed by rSerpinA3n injection (P < 0.05). Besides, WT + OVA group showed more severe in collagen deposition in lung tissues than SerpinA3n-/- + OVA group with increased expression of matrix metallopeptidase-2 (MMP-2), MMP-9, Eotaxin-1, Interleukin 5 (IL-5), IL-13 and IL-4 in lung tissues and deceased IL-10 and Interferon-gamma (IFN-γ) (P < 0.05). Nevertheless, the ameliorating effects of SerpinA3n knockout on OVA-induced asthmatic mice can be reversed by rSerpinA3n. CONCLUSION: SerpinA3n knockout can attenuate airway hyper-reactivity, mitigate inflammatory responses and reduce collagen deposition in lung tissues of neonatal mice with asthma.


Assuntos
Proteínas de Fase Aguda/metabolismo , Asma/metabolismo , Colágeno/metabolismo , Inflamação/metabolismo , Ovalbumina/farmacologia , Serpinas/metabolismo , Animais , Animais Recém-Nascidos , Asma/sangue , Asma/induzido quimicamente , Asma/imunologia , Modelos Animais de Doenças , Feminino , Inflamação/sangue , Inflamação/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout
3.
Int J Biol Macromol ; 177: 422-429, 2021 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-33631260

RESUMO

We isolated and purified a pectin from Portulaca oleracea L. (P. oleracea), and analysed its structure by high-performance size exclusion chromatography (HPSEC), high-performance liquid chromatography (HPLC), gas chromatograph-mass spectrometer (GC-MS), fourier transform infrared spectroscopy (FT-IR), and 1H, 13C nuclear magnetic resonance spectroscopy (NMR). The data indicated that this pectin (designated as POPW-HG) was a linear non-esterified homogalacturonan, which is unique in plants; its molecular weight was around 41.2 kDa. Meanwhile, POPW-HG as an adjuvant was evaluated in the mice immunized with OVA subcutaneously. OVA-specific antibody titres from the sera of immunized mice were tested by ELISA. It showed that POPW-HG significantly enhanced OVA-specific antibody titres (IgG, IgG1, and IgG2b) (p < 0.05) in a dose-dependent manner in the OVA-immunized mice, preliminarily indicating POPW-HG could increase an antibody response, Th1 and Th2 immune response. In addition, the ratio of IgG1/IgG2b suggested POPW-HG induced a Th2-biased response in the OVA-immunized mice. The results demonstrated POPW-HG could be a potential adjuvant candidate in vaccines.


Assuntos
Adjuvantes Imunológicos , Imunização , Ovalbumina/farmacologia , Pectinas , Portulaca/química , Adjuvantes Imunológicos/química , Adjuvantes Imunológicos/isolamento & purificação , Adjuvantes Imunológicos/farmacologia , Animais , Camundongos , Pectinas/química , Pectinas/isolamento & purificação , Pectinas/farmacologia
4.
J Leukoc Biol ; 109(1): 223-232, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-32745316

RESUMO

The efficacy of certain vaccines is improved by the use of adjuvants. Nowadays, the development of new, effective, and safe adjuvants that stimulate the innate immune response is researched. In this context, medicinal plants appear as a suitable alternative. Minthostachys verticillata essential oil (EO) has demonstrated the ability to modulate mechanisms of the innate immune response. Thus, the present work aimed to evaluate the EO adjuvant effect on humoral and cellular immunity, coadministered with OVA as antigen. The chemical analysis of EO by gas chromatography-mass spectrometry revealed a predominant pulegone-menthone chemotype. EO (1.25, 2.5, or 5.0 mg/ml) did not alter the viability of murine fibroblasts (3T3 cell line) neither showed signs of toxicity in Balb/c mice inoculated subcutaneously. The serum of mice immunized with OVA + EO showed increased levels of anti-OVA-specific antibodies of IgG1 subclass compared with the mice immunized with OVA alone revealing an adjuvant effect of EO. The delayed type hypersensitivity showed that the combination OVA + Al(OH)3  + EO was the best to induce a cellular immune response that extended until 48 h postinjection of OVA. M. verticillata EO appears as a new, safe, and effective adjuvant, which should continue to be studied for their possible future incorporation into vaccine formulations.


Assuntos
Adjuvantes Imunológicos/farmacologia , Lamiaceae/imunologia , Óleos Voláteis/farmacologia , Ovalbumina/imunologia , Óleos Vegetais/farmacologia , Hidróxido de Alumínio/imunologia , Hidróxido de Alumínio/farmacologia , Animais , Imunidade Celular/efeitos dos fármacos , Imunidade Celular/imunologia , Imunoglobulina G/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/farmacologia
5.
Integr Zool ; 16(2): 270-279, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32627954

RESUMO

When the integrity of airway epithelium is destroyed, the ordered airway barrier no longer exists and increases sensitivity to viral infections and allergens, leading to the occurrence of airway inflammation such as asthma. Here, we found that galectin-7 transgenic(+) mice exhibited abnormal airway structures as embryos and after birth. These abnormalities included absent or substantially reduced pseudostratified columnar ciliated epithelium and increased monolayer cells with irregular arrangement and widening of intercellular spaces. Moreover, airway tissue from galectin-7 transgenic(+) mice showed evidence of impaired cell-cell junctions and decreased expression of zonula occludens-1(ZO-1) and E-cadherin. When treated with respiratory syncytial virus (RSV) or ovalbumin (OVA), galectin-7 transgenic(+) mice developed substantially increased bronchial epithelial detachment and apoptosis, airway smooth muscle and basement membrane thickening, and enhanced airway responsiveness. We found that Galectin-7 localized in the cytoplasm and nucleus of bronchial epithelial cells, and that increased apoptosis was mediated through mitochondrial release of cytochrome c and upregulated JNK1 activation and expression of caspase-3 in galectin-7 Tg(+) mice. These findings suggested that Galectin-7 causes airway structural defects and destroys airway epithelium barrier, which predispose the airways to RSV or OVA-induced epithelial apoptosis, injury, and other asthma responses.


Assuntos
Asma/fisiopatologia , Células Epiteliais/efeitos dos fármacos , Galectinas/metabolismo , Animais , Apoptose , Brônquios/citologia , Brônquios/efeitos dos fármacos , Brônquios/virologia , Caderinas/metabolismo , Modelos Animais de Doenças , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Células Epiteliais/virologia , Galectinas/genética , Camundongos Transgênicos , Ovalbumina/farmacologia , Ratos Sprague-Dawley , Infecções por Vírus Respiratório Sincicial , Vírus Sinciciais Respiratórios , Junções Íntimas/metabolismo
6.
Biochem Biophys Res Commun ; 534: 714-719, 2021 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-33218687

RESUMO

Aggregation of IgE bound to the high-affinity IgE receptor (FcεRI) by a multivalent antigen induces mast cell activation, while disaggregation of aggregated FcεRI by monomer hapten immediately terminates degranulation mediated by dephosphorylation of Syk and mediates a decrease in intracellular Ca2+ concentration ([Ca2+]i). The actin polymerization state is intimately involved in mast cell activation mediated by FcεRI aggregation. However, the relation between aggregation-disaggregation of FcεRI and actin rearrangement in mast cells is not well understood. The addition of a multivalent antigen rapidly depolymerized actin filaments, while the subsequent addition of monomer hapten rapidly recovered actin polymerization. Whereas cofilin, an actin-severing protein, was temporally dephosphorylated several minutes after a multivalent antigen stimulation and the addition of monomer hapten rapidly increased cofilin phosphorylation level within 30 s. The removal of extracellular Ca2+ instead of monomer hapten addition did not restore cofilin phosphorylation, suggesting that the significant decrease in [Ca2+]i by monovalent hapten was not a critical reason for the actin rearrangement. Additionally, monovalent hapten did not completely reduce [Ca2+]i in mast cells pretreated with jasplakinolide, an inhibitor of actin depolymerization. These results suggest that the multivalent antigen-induced actin depolymerization mediated by cofilin dephosphorylation, and the subsequent addition of monovalent hapten in the F-actin severing state efficiently elicited actin re-polymerization by cofilin phosphorylation.


Assuntos
Fatores de Despolimerização de Actina/metabolismo , Actinas/metabolismo , Mastócitos/metabolismo , Receptores de IgE/metabolismo , Animais , Sinalização do Cálcio , Linhagem Celular , Citocalasina D/farmacologia , Mastócitos/efeitos dos fármacos , Ovalbumina/farmacologia , Faloidina/química , Faloidina/metabolismo , Fosforilação , Polimerização , Ratos , Rodaminas/química , Rodaminas/metabolismo
7.
Artigo em Inglês | MEDLINE | ID: mdl-33373962

RESUMO

Polyunsaturated fatty acids (PUFAs) are present in biological membranes and influence membrane fluidity and immune responses. PUFAs such as 18:2n-6 and 18:3n-3 cannot be synthesized de novo in mammals and are thus called essential fatty acids (EFAs). In addition, PUFAs can be converted to very long-chain PUFAs (VLC-PUFAs), such as arachidonic acid and docosahexaenoic acid, in the body. Although avoiding allergens is an effective strategy for food-allergy patients, the dietary exclusion of several allergens reportedly induces deficiencies in essential nutrients such as PUFAs. In this study, we investigated whether an EFA-deficient (EFAD) diet influenced allergic symptoms in ovalbumin (OVA)-immunized mice. Unexpectedly, no exacerbation of immune responses after OVA-sensitization was observed in mice fed an EFAD diet, and no differences in serum PUFA levels between OVA-immunized and non-immunized mice fed the EFAD diet were detected. However, levels of VLC-PUFAs in the small intestine increased after OVA-sensitization and did not decrease during EFAD diet administration, showing that small intestinal VLC-PUFAs levels were strongly preserved in the food-allergy model mice. Further studies are required to elucidate the mechanisms by which small intestinal VLC-PUFAs are retained in food-allergy model mice.


Assuntos
Ácidos Graxos Essenciais/deficiência , Hipersensibilidade Alimentar/metabolismo , Intestino Delgado/metabolismo , Animais , Modelos Animais de Doenças , Hipersensibilidade Alimentar/patologia , Imunização , Intestino Delgado/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/efeitos adversos , Ovalbumina/farmacologia
8.
Mol Immunol ; 131: 60-67, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33358566

RESUMO

BACKGROUND: Growing evidence shows that enhancer of zeste homolog 2 (EZH2) plays a role in various physiological functions and cancer pathogenesis. However, its contribution to allergic diseases remains controversial. We sought to investigate the role of EZH2 in the pathogenesis of allergic airway inflammation. METHODS: 3-Deazaneplanocin A (DZNep), an indirect inhibitor of EZH2, was administered via intraperitoneal injection in an ovalbumin (OVA)-induced murine model of allergic airway inflammation. The expression of EZH2 in the allergic airway tissues was examined by immunohistochemistry (IHC) and western blot. The inflammatory cell infiltration and the goblet cell hyperplasia in the murine nose and lung were detected by hematoxylin and eosin (H&E) staining and periodic acid-Schiff (PAS) staining. Levels of cytokines, including IL-4, IFN-γ, IL-6, and IL-10, were evaluated in the bronchoalveolar lavage fluid (BALF) using Enzyme-linked immune sorbent assay (ELISA). RESULTS: EZH2 expression was inhibited by DZNep treatment (P < 0.05). The administration of DZNep significantly inhibited the inflammatory cell infiltration (P < 0.0001) and goblet cell hyperplasia (P < 0.001). Moreover, it suppressed the secretion of IL-4 (P < 0.0001) and IL-6 (P < 0.01) in the BALF. CONCLUSIONS: Our findings demonstrate that DZNep attenuates allergic airway inflammation and could be a new therapeutic option for allergic rhinitis and asthma.


Assuntos
Adenosina/análogos & derivados , Anti-Inflamatórios/farmacologia , Hipersensibilidade/tratamento farmacológico , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Pulmão/efeitos dos fármacos , Ovalbumina/farmacologia , Adenosina/farmacologia , Animais , Asma/induzido quimicamente , Asma/tratamento farmacológico , Asma/metabolismo , Líquido da Lavagem Broncoalveolar , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Hipersensibilidade/metabolismo , Inflamação/metabolismo , Pulmão/metabolismo , Camundongos , Camundongos Endogâmicos BALB C
9.
Mol Med Rep ; 22(6): 4909-4918, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33174031

RESUMO

Asthma is a leading allergic disease worldwide, demonstrating an ever­increasing prevalence over the past two decades. Asthma is characterized by allergen­associated airway hyperresponsiveness (AHR) that primarily results from T helper 2 (Th2) cell inflammation, in which dendritic cells (DCs) serve an important role in determining T cell development after encountering an antigen. Atractylodin (ATL), a polyethene alkyne extracted from Atractylodis rhizoma (also known as Cangzhu), has proven effective in treating digestive disorders, rheumatic disease and influenza. In addition, ATL was discovered to alleviate mouse collagen­induced arthritis via regulating DC maturation. The present study aimed to investigate the effect of ATL on asthma given that DCs serve an essential role in Th2­mediated inflammation in asthma. Mouse model of asthma was induced by ovalbumin (OVA). OVA­induced airway hyperresponsiveness (AHR) and inflammatory cells in bronchoalveolar lavage fluid (BALF) were detected. The production of IgE and IgG1 in serum and cytokines in BALF were detected by ELISA. The effects of ATL on dendritic cells maturation and T cell expansion were detected by flow cytometry analysis and 3H­thymidine incorporation. Using a model of OVA­induced asthma, it was demonstrated that ATL ameliorated AHR and decreased the levels of IL­4, IL­5 and IL­13 in bronchoalveolar lavage fluid (BALF), and OVA­specific IgE and IgG1 in the serum. OVA­stimulated splenocytes were used to demonstrated that ATL decreased cell expansion and the production of IL­4, IL­5 and IL­13 in the culture medium. In order to determine the cellular mechanism of ATL in asthma, splenic DCs were isolated and it was subsequently observed that ATL downregulated the expression levels of CD40 and CD80. Furthermore, OVA­stimulated CD4+ T cells were co­cultured with splenic DCs, which revealed that ATL­treated splenic DCs led to impaired cellular proliferation and the production of IL­4, IL­5 and IL­13 in OVA­stimulated T cells. In conclusion, these results indicated that ATL may suppress antigen­specific Th2 responses in an OVA­induced allergic asthma model via regulating DCs. Therefore, ATL may exhibit therapeutic potential in the management of asthma and other allergic diseases presenting with Th2 inflammation.


Assuntos
Asma/tratamento farmacológico , Células Dendríticas/imunologia , Furanos/farmacologia , Alérgenos/metabolismo , Animais , Asma/metabolismo , Hiper-Reatividade Brônquica/imunologia , Líquido da Lavagem Broncoalveolar/citologia , China , Citocinas/metabolismo , Células Dendríticas/efeitos dos fármacos , Modelos Animais de Doenças , Furanos/metabolismo , Imunoglobulina E/sangue , Fatores Imunológicos/imunologia , Inflamação/metabolismo , Interleucina-13/imunologia , Interleucina-4/imunologia , Interleucina-5/imunologia , Pulmão/metabolismo , Pulmão/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/efeitos adversos , Ovalbumina/farmacologia , Linfócitos T Reguladores/imunologia , Células Th2/efeitos dos fármacos , Células Th2/imunologia
10.
Int J Nanomedicine ; 15: 8945-8961, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33223829

RESUMO

Purpose: Isoliquiritigenin (ILQ), an important component of Anti-Asthma Herbal Medicine Intervention (ASHMI), had shown potent anti-asthma effect in vitro in our previous study. However, poor solubility and low bioavailability hindered in vivo application to treat asthma. This study was to develop a novel ILQ loaded self-nanoemulsifying drug delivery system (ILQ-SMEDDS) with enhanced bioavailability. Methods: The optimized SMEDDS formulation was composed of ethyl oleate (oil phase), Tween 80 (surfactant) and PEG400 (co-surfactant) at a mass ratio of 3:6:1. The physiochemical properties of ILQ-SMEDDS, including drug content, globule size, zeta potential, scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, were characterized. And the in vitro release profile, in situ intestinal absorption, in vivo pharmacokinetic parameters and the anti-asthma effect of ILQ suspension and ILQ-SMEDDS were evaluated. Results: The ILQ-SMEDDS had an average globule size of 20.63 ± 1.95 nm with a polydispersity index (PDI) of 0.11 ± 0.03, and its zeta potential was -12.64 ± 2.12 mV. The cumulative release rate of ILQ from ILQ-SMEDDS to the simulated gastrointestinal tract was significantly higher than that of free ILQ suspension. And area under curve with ILQ-SMEDDS was found to be 3.95 times higher than that of ILQ suspension indicating improved bioavailability by SMEDDS. Although ILQ-SMEDDS showed a slight less effective inhibitory effect on eotaxin-1 in human lung fibroblast (HFL-1) cells than free ILQ, in an ovalbumin-induced asthma model, ILQ-SMEDDS exhibited more efficacy than ILQ suspension in improving asthma-associated inflammation, including eosinophil production, ovalbumin-specific immunoglobulin E (OVA-sIgE), interleukin 4 (IL 4), interleukin 5 (IL 5) and interferon-γ (IFN-γ). Even the low dose of ILQ-SMEDDS group (10 mg/kg) showed better anti-asthma effect than that of the ILQ suspension group (20 mg/kg). Conclusion: Compared with ILQ suspension, ILQ-SMEDDS showed significantly improved bioavailability and anti-asthma effect, revealing its potential as a favorable pharmaceutical agent for treating asthma.


Assuntos
Asma/tratamento farmacológico , Chalconas/farmacocinética , Portadores de Fármacos/química , Nanoestruturas/química , Ovalbumina/farmacologia , Administração Oral , Animais , Asma/induzido quimicamente , Disponibilidade Biológica , Chalconas/administração & dosagem , Chalconas/química , Chalconas/uso terapêutico , Emulsões , Humanos , Absorção Intestinal , Masculino , Polietilenoglicóis/química , Polissorbatos/química , Solubilidade , Tensoativos/química
11.
Front Immunol ; 11: 561724, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33224135

RESUMO

Endogenous redox systems not only counteract oxidative damage induced by high levels of hydroxyl radicals (OH·) under pathological conditions, but also shape redox signaling as a key player in the regulation of physiological processes. Second messengers like hydrogen peroxide and nitric oxide, as well as redox enzymes of the Thioredoxin (Trx) family, including Trxs, glutaredoxins (Grxs), and peroxiredoxins (Prxs) modulate reversible, oxidative modifications of proteins. Thereby redox regulation is part of various cellular processes such as the immune response and Trx proteins have been linked in different disorders including inflammatory diseases. Here, we have analyzed the protein distribution of representative oxidoreductases of the Trx fold protein family-Trx1, Grx1, Grx2, and Prx2-in a murine model of allergic asthma bronchiale, as well as their potential therapeutic impact on type-2 driven airway inflammation. Ovalbumin (OVA) sensitization and challenge using the type-2 prone Balb/c mouse strain resulted in increased levels of all investigated proteins in distinct cellular patterns. While concomitant treatment with Grx1 and Prx2 did not show any therapeutic impact on the outcome of the disease, Grx2 or Trx1 treatment before and during the OVA challenge phase displayed pronounced protective effects on the manifestation of allergic airway inflammation. Eosinophil numbers and the type-2 cytokine IL-5 were significantly reduced while lung function parameters profoundly improved. The number of macrophages in the bronchoalveolar lavage (BAL) did not change significantly, however, the release of nitric oxide that was linked to airway inflammation was successfully prevented by enzymatically active Grx2 ex vivo. The Grx2 Cys-X-X-Ser mutant that facilitates de-/glutathionylation, but does not catalyze dithiol/disulfide exchange lost the ability to protect from airway hyper reactivity and to decrease NO release by macrophages, however, it reduced the number of infiltrating immune cells and IL-5 release. Altogether, this study demonstrates that specific redox proteins and particular enzyme activities protect against inflammatory damage. During OVA-induced allergic airway inflammation, administration of Grx2 exerts beneficial and thus potentially therapeutic effects.


Assuntos
Asma/sangue , Asma/tratamento farmacológico , Glutarredoxinas/administração & dosagem , Glutarredoxinas/sangue , Substâncias Protetoras/administração & dosagem , Animais , Asma/induzido quimicamente , Modelos Animais de Doenças , Feminino , Humanos , Inflamação/tratamento farmacológico , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Óxido Nítrico/metabolismo , Ovalbumina/farmacologia , Oxirredução/efeitos dos fármacos , Células RAW 264.7 , Proteínas Recombinantes/administração & dosagem , Transdução de Sinais/efeitos dos fármacos , Tiorredoxinas/administração & dosagem
12.
Biomed Res Int ; 2020: 7407016, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32953887

RESUMO

Pingchuan formula (PCF) was created by Professor Yu Jianer. The purpose of this study was to investigate the effect of PCF on dendritic cells (DCs) and toll-like receptors (TLRs) in initiating immunity. A bronchial asthma BALB/c mouse model was established using an OVA excitation method. PCF was immediately administered by gavage after the first excitation. After 7 d, hematoxylin and eosin (HE) staining was used to observe the pathological changes in the asthma model. Eosinophil infiltration and concentrations of IL-4, IFN-r, IL-12, and IFN-α in BALF were determined by enzyme-linked immunosorbent assay (ELISA). Real-time PCR was used to determine mRNA levels of IL-12 and IFN-α. Protein expression levels of ERK, Toll-2, IDO, and Toll-9 were measured by immunoblot. HE and ELISA showed that PCF could improve lung pathological changes and significantly decrease the concentration of IL-4 in BALF. Moreover, PCF could increase IL-12, IFN-α, and IFN-r in BALF. Real-time PCR and western blot showed that PCF restored the DCs and TLRs in initiating immunity. In summary, this study found that PCF can improve the pathological changes and reduce the symptoms of asthma in a BALB/c mouse model. It can facilitate the initiation of immunity by restoring the DCs and TLRs.


Assuntos
Asma/tratamento farmacológico , Células Dendríticas/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Receptores Toll-Like/metabolismo , Animais , Asma/induzido quimicamente , Asma/metabolismo , Líquido da Lavagem Broncoalveolar , Células Dendríticas/metabolismo , Modelos Animais de Doenças , Imunidade/efeitos dos fármacos , Interferon-alfa/metabolismo , Interferon gama/metabolismo , Interleucina-12/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/farmacologia , RNA Mensageiro/metabolismo
13.
Med Sci Monit ; 26: e923358, 2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32868754

RESUMO

BACKGROUND Allergic rhinitis (AR) is a prevalent atopic disorder caused by immune imbalance. Chlorogenic acid (CGA) has antibacterial, antiviral, antioxidative and immunoregulatory effects, but its role in anaphylactic disease remains unclear. The current study aimed to investigate the function of CGA in AR. MATERIAL AND METHODS AR mice models were induced with ovalbumin (OVA) by orally administrating the mice with 50 mg/kg (L-CGA), 100 mg/kg (M-CGA), or 200 mg/kg (H-CGA) of CGA. The number of nasal rubbings and sneezes was recorded. Afterward, the mice were sacrificed for the collection of blood, nasal lavage fluid (NALF), and nasal tissues. The cells in NALF were counted by hemocytometer and stained by Diff-Quick. Nasal mucosa was observed by H&E staining. ELISA testing was conducted for detecting the levels of anti-OVA antibodies and Th1/Th2-related cytokine. Quantitative real-time polymerase chain reaction experiments were conducted to determine mRNA expressions of Th1/Th2-related cytokines. RESULTS In the OVA-induced AR mice, CGA treatment reduced nasal rubbing and sneezing, and also suppressed the number of total cells, eosinophils, neutrophils, lymphocytes, macrophages, and epithelial cells in NALF. OVA-induced up-regulation of nasal mucosa thickness was inhibited by CGA, and the effects of OVA on IgE, IgG1, and IgG2a were reversed by CGA. The regulatory effects of OVA on mRNA expressions and levels of Th1/Th2-related cytokines were abolished by CGA treatment in AR mice. CONCLUSIONS CGA can alleviate allergic inflammatory responses through regulating Th1/Th2 balance in OVA-induced allergic rhinitis mice.


Assuntos
Ácido Clorogênico/uso terapêutico , Ovalbumina/farmacologia , Rinite Alérgica/tratamento farmacológico , Equilíbrio Th1-Th2/efeitos dos fármacos , Animais , Ácido Clorogênico/farmacologia , Citocinas/metabolismo , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Eosinófilos/efeitos dos fármacos , Eosinófilos/metabolismo , Mediadores da Inflamação/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Mucosa Nasal/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Rinite Alérgica/imunologia
14.
Mol Biol Rep ; 47(5): 3735-3744, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32378169

RESUMO

In this study, a mice model of obesity-asthma was established. We investigated the correlation between oxidative stress and NF-κB signaling pathway in the lung tissues, together with the effects of acetylcysteine. The animals were fed on a high-fat diet, and then ovalbumin (OVA) sensitization was utilized to establish the obesity-asthma model. N-acetylcysteine was used to treat asthma, animals treated with budesonide served as control. The malondialdehyde (MDA) in the lung tissues was determined, together with the activity of glutathione (GSH). EMAS assay was utilized to measure the nuclear factor-κB-P65 (NF-κB-P65) in lung tissues. Western blot analysis was performed to determine the expression of inhibitor kappa B-α (IκB-α) and inhibitor kappa B kinase-ß (IKK-ß). The MDA in the asthma groups showed significantly elevation (P < 0.01), and the GSH showed significant decrease (P < 0.01), especially in the obesity-asthma group. The efficiency of N-acetylcysteine was superior to that of the budesonide in the decline of MDA and elevation of GSH (P < 0.01). In both asthma groups, the expression of IKK-ß and transcription of NF-κB-P65 in the lung tissues showed significant elevation (P < 0.01), and IκB-α showed significant decline (P < 0.01), especially in the obesity-asthma group. There was decline of IKK-ß and NF-κB-P65 and elevation of IκB-α in the N-acetylcysteine group, which was even significantly in the Budesonide group (P < 0.01). There was a positive correlation between MDA and NF-κB activation in the lung tissues in all the asthma groups and treatment groups (P < 0.05). Obesity-asthma mice showed higher oxidative stress and activation of NF-κB compared with that of the asthma mice. There was a positive correlation between MDA and NF-κB activation in the lung tissues in the asthma groups. N-acetylcysteine was more effective in reducing the oxidative stress compared to the budesonide.


Assuntos
Asma/metabolismo , Obesidade/metabolismo , Estresse Oxidativo/fisiologia , Acetilcisteína/farmacologia , Animais , Asma/fisiopatologia , Feminino , Glutationa/análise , Quinase I-kappa B/metabolismo , Pulmão/citologia , Pulmão/metabolismo , Malondialdeído/análise , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Obesos , Inibidor de NF-kappaB alfa/metabolismo , NF-kappa B/metabolismo , Obesidade/fisiopatologia , Ovalbumina/farmacologia , Fosforilação/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismo
15.
PLoS One ; 15(5): e0226233, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32379832

RESUMO

Allergic asthma is the most common phenotype of the pathology, having an early-onset in childhood and producing a Th2-driven airways remodeling process that leads to symptoms and pathophysiological changes. The avoidance of aeroallergen exposure in early life has been shown to prevent asthma, but without repeated success and with the underlying preventive mechanisms at the beginning of asthma far to be fully recognized. In the present study, we aimed to evaluate if neonatal LPS-induced boost in epithelial host defenses contribute to prevent OVA-induced asthma in adult mice. To this, we focused on the response of bronchiolar club cells (CC), which are highly specialized in maintaining the epithelial homeostasis in the lung. In these cells, neonatal LPS administration increased the expression of TLR4 and TNFα, as well as the immunodulatory/antiallergic proteins: club cell secretory protein (CCSP) and surfactant protein D (SP-D). LPS also prevented mucous metaplasia of club cells and reduced the epidermal growth factor receptor (EGFR)-dependent mucin overproduction, with mice displaying normal breathing patterns after OVA challenge. Furthermore, the overexpression of the epithelial Th2-related molecule TSLP was blunted, and normal TSLP and IL-4 levels were found in the bronchoalveolar lavage. A lower eosinophilia was detected in LPS-pretreated mice, along with an increase in phagocytes and regulatory cells (CD4+CD25+FOXP3+ and CD4+IL-10+), together with higher levels of IL-12 and TNFα. In conclusion, our study demonstrates stable asthma-preventive epithelial effects promoted by neonatal LPS stimulation, leading to the presence of regulatory cells in the lung. These anti-allergic dynamic mechanisms would be overlaid in the epithelium, favored by an adequate epidemiological environment, during the development of asthma.


Assuntos
Asma/imunologia , Bronquíolos/efeitos dos fármacos , Bronquíolos/imunologia , Citocinas/metabolismo , Epitélio/imunologia , Imunidade Inata , Lipopolissacarídeos/imunologia , Lipopolissacarídeos/farmacologia , Alérgenos/imunologia , Animais , Animais Recém-Nascidos , Asma/prevenção & controle , Modelos Animais de Doenças , Epitélio/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia , Ovalbumina/farmacologia , Linfócitos T Reguladores/efeitos dos fármacos , Linfócitos T Reguladores/imunologia
16.
J Cell Physiol ; 235(12): 9464-9473, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-32394447

RESUMO

There has been a marked increase in life-threatening food allergy (FA). One hypothesis is that changes in bacterial communities may be key to FA. To better understand how gut microbiota regulates FA in humans, we established a mouse model with FA induced by ovalbumin. We found that the mice with FA had abnormal bacterial composition, accompanied by increased immunoglobulin G, immunoglobulin E, and interleukin-4/interferon-γ, and there existed a certain coherence between them. Interestingly, Bifidobacterium breve M-16V may alter the gut microbiota to alleviate the allergy symptoms by IL-33/ST2 signaling. Our results indicate that gut microbiota is essential for regulating FA to dietary antigens and demonstrate that intervention in bacterial community regulation may be therapeutically related to FA.


Assuntos
Hipersensibilidade Alimentar/tratamento farmacológico , Interleucina-33/metabolismo , Probióticos/farmacologia , Transdução de Sinais/efeitos dos fármacos , Animais , Bifidobacterium breve/efeitos dos fármacos , Bifidobacterium breve/imunologia , Microbioma Gastrointestinal/efeitos dos fármacos , Microbioma Gastrointestinal/imunologia , Camundongos Endogâmicos BALB C , Ovalbumina/farmacologia , Transdução de Sinais/imunologia
17.
Elife ; 92020 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-32412411

RESUMO

Millions of naïve T cells with different TCRs may interact with a peptide-MHC ligand, but very few will activate. Remarkably, this fine control is orchestrated using a limited set of intracellular machinery. It remains unclear whether changes in stimulation strength alter the programme of signalling events leading to T cell activation. Using mass cytometry to simultaneously measure multiple signalling pathways during activation of murine CD8+ T cells, we found a programme of distal signalling events that is shared, regardless of the strength of TCR stimulation. Moreover, the relationship between transcription of early response genes Nr4a1 and Irf8 and activation of the ribosomal protein S6 is also conserved across stimuli. Instead, we found that stimulation strength dictates the rate with which cells initiate signalling through this network. These data suggest that TCR-induced signalling results in a coordinated activation program, modulated in rate but not organization by stimulation strength.


Assuntos
Linfócitos T CD8-Positivos/efeitos dos fármacos , Ativação Linfocitária/efeitos dos fármacos , Ovalbumina/farmacologia , Receptores de Antígenos de Linfócitos T/agonistas , Transdução de Sinais/efeitos dos fármacos , Animais , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Células Cultivadas , Feminino , Citometria de Fluxo , Fatores Reguladores de Interferon/genética , Fatores Reguladores de Interferon/metabolismo , Cinética , Ligantes , Masculino , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/genética , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/metabolismo , Fragmentos de Peptídeos/farmacologia , Fosforilação , Receptores de Antígenos de Linfócitos T/metabolismo , Proteína S6 Ribossômica/metabolismo , Análise de Célula Única
18.
Chemosphere ; 250: 126244, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32113099

RESUMO

To investigate the effect of gestational and lactational nonylphenol (NP) exposure on airway inflammation in ovalbumin (OVA)-induced asthmatic pups. Dams were gavaged with NP at dose levels of 25 mg/kg/day (low dose), 50 mg/kg/day (middle dose), 100 mg/kg/day (high dose) and groundnut oil alone (vehicle control) respectively from gestational day 7 to postnatal day 21. The results showed that the NP content in the lung tissues of pups in the 100 mg/kg NP group was significantly higher than that of the control group (P = 0.004). In the 100 mg/kg NP group, the infiltration of lymphocytes and eosinophils with thicken smooth muscle layer and inflammatory cells in the lumen were observed in the lung tissues of pups. Osmiophilic lamellar bodies were found in the cytoplasm of type II epithelial cells; mitochondria were clearly swollen. Compared with the control group, the levels of interleukin-4 (IL-4) in BALF (P = 0.042) and ovalbumin-specific serum immunoglobulin E (OVA-sIgE) (P = 0.005) in the OVA group were significantly higher. 25 mg/kg NP-OVA co-exposure synergistically decreased nuclear factor-κB (NF-κB) mRNA expression in the lung tissues of pups; Exposure to 50 mg/kg NP combined with OVA antagonized the increased expression of high mobility group box 1 (HMGB1) mRNA in the lung tissue. The combined exposure to 50 mg/kg NP and OVA synergistically increased HMGB1 protein expression in the lung tissues. 25 mg/kg NP-OVA co-exposure antagonized the increased nuclear factor-κB (NF-κB) protein expression in the lung tissues. There was a positive correlation between NP content and HMGB1 protein expression in the lung tissue of asthmatic pups (r = 0.602, P < 0.001). In conclusion, gestational and lactational exposure to 100 mg/kg NP in maternal rats exacerbated airway inflammation in OVA-induced asthmatic pups, and there is an interactive effect between NP and OVA. When the perinatal rats were exposed to 100 mg/kg NP, the levels of HMGB1 and NF-κB in the lung tissues of OVA-induced asthmatic pups were increased.


Assuntos
Poluentes Ambientais/toxicidade , Fenóis/toxicidade , Animais , Asma/metabolismo , Modelos Animais de Doenças , Feminino , Inflamação/metabolismo , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Camundongos Endogâmicos BALB C , NF-kappa B/metabolismo , Ovalbumina/farmacologia , Gravidez , Ratos , Testes de Toxicidade
19.
Sci Rep ; 10(1): 4099, 2020 03 05.
Artigo em Inglês | MEDLINE | ID: mdl-32139713

RESUMO

Exposure to chloromethylisothiazolinone/methylisothiazolinone (CMIT/MIT) has been associated with allergic contact dermatitis and occupational asthma. Despite this association however, no study has investigated the effects of CMIT/MIT exposure on the development of atopic dermatitis (AD). This study was conducted to investigate the influence of epicutaneous exposure to CMIT/MIT on AD in a mouse model and the underlying biological mechanisms. BALB/C mice were exposed to CMIT/MIT for 3 weeks and AD was developed using ovalbumin (OVA) epidermal sensitization. CMIT/MIT epicutaneous exposure in normal mice significantly enhanced AD-like phenotypes (e.g., transepidermal water loss, clinical score, total serum immunoglobulin E level and infiltration of inflammatory cells). In addition, CMIT/MIT exposure significantly augmented the mRNA expression level of T helper (Th) 2-related cytokines (thymic stromal lymphopoietin, interleukin (IL)-6 and IL-13), Th2 chemokine (chemokine (C-C motif) ligand 17) and the population of CD4+IL-4+ cells in the skin. Moreover, mice exposed to CMIT/MIT in the OVA challenge had greater AD-like phenotypes, higher IL-4 and IL-17A skin mRNA expression levels, and a larger population of CD4+IL-4+- and IL-17A+-producing cells in the skin-draining lymph nodes. Our current findings in a mouse model thus suggest that CMIT/MIT exposure may cause AD symptoms through the dysregulation of Th2/Th17-related immune responses.


Assuntos
Dermatite Atópica/induzido quimicamente , Poluentes Ambientais/toxicidade , Fatores Imunológicos/toxicidade , Células Th17/imunologia , Células Th2/imunologia , Tiazóis/toxicidade , Animais , Dermatite Atópica/imunologia , Dermatite Atópica/patologia , Modelos Animais de Doenças , Feminino , Imunidade , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/farmacologia , Pele/efeitos dos fármacos , Pele/patologia
20.
Cell Transplant ; 29: 963689720913254, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32216447

RESUMO

Mesenchymal stem cell (MSC) research has demonstrated the potential of these cells to modulate lung inflammatory processes and tissue repair; however, the underlying mechanisms and treatment durability remain unknown. Here, we investigated the therapeutic potential of human bone marrow-derived MSCs in the inflammatory process and pulmonary remodeling of asthmatic BALB/c mice up to 14 d after transplantation. Our study used ovalbumin to induce allergic asthma in male BALB/c mice. MSCs were injected intratracheally in the asthma groups. Bronchoalveolar lavage fluid (BALF) was collected, and cytology was performed to measure the total protein, hydrogen peroxide (H2O2), and proinflammatory (IL-5, IL-13, and IL-17A) and anti-inflammatory (IL-10) interleukin (IL) levels. The lungs were removed for the histopathological evaluation. On day zero, the eosinophil and lymphochte percentages, total protein concentrations, and IL-13 and IL-17A levels in the BALF were significantly increased in the asthma group, proving the efficacy of the experimental model of allergic asthma. On day 7, the MSC-treated group exhibited significant reductions in the eosinophil, lymphocyte, total protein, H2O2, IL-5, IL-13, and IL-17A levels in the BALF, while the IL-10 levels were significantly increased. On day 14, the total cell numbers and lymphocyte, total protein, IL-13, and IL-17A levels in the BALF in the MSC-treated group were significantly decreased. A significant decrease in airway remodeling was observed on days 7 and 14 in almost all bronchioles, which showed reduced inflammatory infiltration, collagen deposition, muscle and epithelial thickening, and mucus production. These results demonstrate that treatment with a single injection of MSCs reduces the pathophysiological events occurring in an experimental model of allergic asthma by controlling the inflammatory process up to 14 d after transplantation.


Assuntos
Medula Óssea/metabolismo , Pulmão/efeitos dos fármacos , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/metabolismo , Pneumonia/tratamento farmacológico , Animais , Anti-Inflamatórios/farmacologia , Líquido da Lavagem Broncoalveolar/citologia , Citocinas/metabolismo , Modelos Animais de Doenças , Humanos , Pulmão/patologia , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/efeitos dos fármacos , Camundongos Endogâmicos BALB C , Ovalbumina/farmacologia
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