RESUMO
Inflammatory bowel disease (IBD) is a chronic persistent intestinal disorder, with ulcerative colitis and Crohn's disease being the most common. However, the physio-pathological development of IBD is still unknown. Therefore, research on the etiology and treatment of IBD has been conducted using a variety of approaches. Short-chain fatty acids such as 3-hydroxybutyrate (3-HB) are known to have various physiological activities. In particular, the production of 3-HB by the intestinal microflora is associated with the suppression of various inflammatory diseases. In this study, we investigated whether poly-D-3-hydroxybutyric acid (PHB), a polyester of 3-HB, is degraded by intestinal microbiota and works as a slow-release agent of 3-HB. Further, we examined whether PHB suppresses the pathogenesis of IBD models. As long as a PHB diet increased 3-HB concentrations in the feces and blood, PHB suppressed weight loss and histological inflammation in a dextran sulfate sodium-induced IBD model. Furthermore, PHB increased the accumulation of regulatory T cells in the rectum without affecting T cells in the spleen. These results indicate that PHB has potential applications in treating diseases related to the intestinal microbiota as a sustained 3-HB donor. We show for the first time that biodegradable polyester exhibits intestinal bacteria-mediated bioactivity toward IBD. The use of bioplastics, which are essential materials for sustainable social development, represents a novel approach to diseases related to dysbiosis, including IBD.
Assuntos
Doenças Inflamatórias Intestinais , Linfócitos T Reguladores , Humanos , Ácido 3-Hidroxibutírico/farmacologia , Ácido 3-Hidroxibutírico/metabolismo , Linfócitos T Reguladores/metabolismo , Regulação para Cima , Doenças Inflamatórias Intestinais/tratamento farmacológico , Doenças Inflamatórias Intestinais/metabolismo , Hidroxibutiratos/farmacologia , PoliésteresRESUMO
This study investigated the immunotoxic effects of the mycotoxin nivalenol (NIV) using antigen-presenting cells and a mouse model of atopic dermatitis (AD). In vitro experiments were conducted using a mouse macrophage cell line (RAW 264.7) and mouse dendritic cell line (DC 2.4). After cells were exposed to NIV (0.19-5 µmol) for 24 h, the production of pro-inflammatory cytokines (IL-1ß, IL-6, and TNFα) was quantified. To further investigate the inflammatory cytokine production pathway, the possible involvement of mitogen-activated protein kinase (MAPK) pathways, such as ERK1/2, p-38, and JNK, in NIV exposure was analyzed using MAPK inhibitors and phosphorylation analyses. In addition, the pro-inflammatory effects of oral exposure to NIV at low concentrations (1 or 5 ppm) were evaluated in an NC/Nga mouse model of hapten-induced AD. In vitro experiments demonstrated that exposure to NIV significantly enhanced the production of TNFα. In addition, it also directly induced the phosphorylation of MAPK, indicated by the inhibition of TNFα production following pretreatment with MAPK inhibitors. Oral exposure to NIV significantly exacerbated the symptoms of AD, including a significant increase in helper T cells and IgE-produced B cells in auricular lymph nodes and secretion of pro-inflammatory cytokines, such as IL-4, IL-5, and IL-13, compared with the vehicle control group. Our findings indicate that exposure to NIV directly enhanced the phosphorylation of ERK1/2, p-38, and JNK, resulting in a significant increase in TNFα production in antigen-presenting cells, which is closely related to the development of atopic dermatitis.
Assuntos
Citocinas , Dermatite Atópica , Tricotecenos , Animais , Dermatite Atópica/induzido quimicamente , Dermatite Atópica/imunologia , Tricotecenos/toxicidade , Tricotecenos/administração & dosagem , Camundongos , Administração Oral , Citocinas/metabolismo , Células RAW 264.7 , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Células Apresentadoras de Antígenos/efeitos dos fármacos , Células Apresentadoras de Antígenos/imunologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Modelos Animais de Doenças , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/metabolismo , Células Dendríticas/imunologia , Fosforilação , Masculino , Fator de Necrose Tumoral alfa/metabolismo , FemininoRESUMO
This study aimed to determine whether oral fumonisin exposure contributes to the development of psoriasis. Oral administration of fumonisin B1 (FB1, 0.1 mg/kg) or fumonisin B2 (FB2, 0.1 mg/kg) was conducted for 10 days, in addition to the induction of psoriatic symptoms through topical application of 5% imiquimod cream from day 6 to day 10 (5 days) in female BALB/c mice. The results demonstrated that oral administration of FB2 significantly exacerbated psoriatic symptoms, including skin thickness, itching behavior, transepidermal water loss, immune cell infiltration in the dermis, and proinflammatory cytokine production. However, no changes were observed following exposure to FB1. Our results confirm that oral exposure to FB2 adversely affects the pathogenesis of psoriasis by increasing skin thickness and impairing barrier function.
Assuntos
Fumonisinas , Imiquimode , Camundongos Endogâmicos BALB C , Psoríase , Animais , Psoríase/induzido quimicamente , Psoríase/patologia , Psoríase/metabolismo , Imiquimode/efeitos adversos , Fumonisinas/toxicidade , Camundongos , Feminino , Administração Oral , Pele/efeitos dos fármacos , Pele/metabolismo , Pele/patologia , Citocinas/metabolismo , Modelos Animais de DoençasRESUMO
Epidermal tissues play vital roles in maintaining homeostasis and preventing the dysregulation of the cutaneous barrier. Sphingomyelin (SM), a sphingolipid synthesized by sphingomyelin synthase (SMS) 1 and 2, is involved in signal transduction via modulation of lipid-raft functions. Though the implications of SMS on inflammatory diseases have been reported, its role in dermatitis has not been clarified. In this study, we investigated the role of SM in the cutaneous barrier using a dermatitis model established by employing Sgms1 and 2 deficient mice. SM deficiency impaired the cutaneous inflammation and upregulated signal transducer and activator of transcription 3 (STAT3) phosphorylation in epithelial tissues. Furthermore, using mouse embryonic fibroblast cells, the sensitivity of STAT3 to Interleukin-6 stimulation was increased in Sgms-deficient cells. Using tofacitinib, a clinical JAK inhibitor, the study showed that SM deficiency might participate in STAT3 phosphorylation via JAK activation. Overall, these results demonstrate that SM is essential for maintaining the cutaneous barrier via the STAT3 pathway, suggesting SM could be a potential therapeutic target for dermatitis treatment.
Assuntos
Fator de Transcrição STAT3/fisiologia , Pele/metabolismo , Esfingomielinas/fisiologia , Animais , Células Cultivadas , Dermatite/tratamento farmacológico , Dermatite/etiologia , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Fosforilação , Transdução de Sinais/fisiologia , Esfingomielinas/uso terapêutico , Transferases (Outros Grupos de Fosfato Substituídos)/fisiologiaRESUMO
Contamination with fumonisins produced by Fusarium spp. is rapidly growing in both developing and developed countries. The purpose of this study was to determine whether oral exposure to fumonisin contributed to the development of allergic diseases. We initially examined the immunotoxic potential of short-term, oral administration of fumonisin B1 (FB1, 1 mg/kg) and fumonisin B2 (FB2, 1 mg/kg), both naturally occurring fumonisins, using a BALB/c mouse model of allergic contact dermatitis and Dermatophagoides farina-induced asthma. Using an NC/nga mouse model of atopic dermatitis (AD), we evaluated the adverse effects of subchronic oral exposure to low concentrations of FB2 (2 or 200 µg/kg). Finally, we explored the influence of FB2 on regulatory T cell proliferation and function in mesenteric lymph nodes after 1-week oral exposure to FB2 in BALB/c mice. Oral exposure to FB2 markedly exacerbated the symptoms of allergy, including skin thickness, histological evaluation, immunocyte proliferation, and proinflammatory cytokine production, although no change was observed following exposure to FB1. Furthermore, oral exposure to low concentrations of FB2 considerably exacerbated the AD scores, skin thickness, transepidermal water loss, histological features, and proinflammatory cytokine production. The aggravated allergic symptoms induced by oral exposure to FB2 could be attributed to the direct inhibition of IL-10 production by regulatory T cells in mesenteric lymph nodes. Our findings indicate that the recommended maximum fumonisin level should be reconsidered based on the potential for allergy development.
Assuntos
Dermatite Alérgica de Contato , Fumonisinas , Animais , Camundongos , Fumonisinas/toxicidade , Interleucina-10 , Linfócitos T Reguladores , LinfonodosRESUMO
Ozone gas is widely used in hospitals as well as homes to control COVID-19 infection owing to its cost-effectiveness. Safety standard value and the tolerable value of ozone gas are set at 0.05 ppm and 0.1 ppm, respectively, in developed countries; however, this value was principally determined for healthy individuals, and the risks associated with ozone gas inhalation in patients with pulmonary diseases remains unknown. Recently, we demonstrated that 0.1 ppm ozone gas exposure significantly aggravates the symptoms of acute lung injury in mice. In the present study, we further examined the influence of ≤ 0.1 ppm ozone gas exposure on percutaneous oxygen saturation (SpO2) and pro-inflammatory responses in a mouse model of asthma. Female BALB/c mice were subjected to repetitive intranasal sensitization of Dermatophagoides farinae to generate a mouse model of asthma. Inhalation exposure of ozone gas (0.1, 0.03, 0.01 ppm), generated using an ultraviolet lamp, was performed for five consecutive days immediately before the final sacrifice. There were no abnormal findings in control mice exposed to 0.1 ppm ozone; however, 0.1 ppm ozone exposure significantly reduced the SpO2 level in asthmatic mice. Histological evaluation and gene expression analysis revealed that pro-inflammatory cytokine levels were significantly increased in mice exposed to 0.1 ppm ozone, indicating that 0.1 ppm ozone exposure affects the development of asthma symptoms. Notably, 0.03 and 0.01 ppm ozone exposure did not have any effects even in asthmatic mice. Our findings indicate that the tolerable level of ozone gas should be adjusted for individuals based on a history of respiratory disorders.
Assuntos
Asma , COVID-19 , Ozônio , Humanos , Feminino , Animais , Camundongos , Dermatophagoides farinae , Saturação de Oxigênio , Asma/induzido quimicamente , Modelos Animais de Doenças , Ozônio/toxicidade , PulmãoRESUMO
Citrinin, a mycotoxin produced by Penicillium citrinum and Penicillium verrucosum, mainly contaminates cereals. The aim of study was to investigate the novel immunoreactive effect of citrinin using a mouse model of psoriasis. A mouse model of psoriasis was generated by topical application of 5% imiquimod in female BALB/c mice. Standard rodent diet and rice samples with 3 ppm of citrinin were mixed to obtain a final citrinin concentration of 0.3 ppm, and a citrinin-contaminated diet was fed to mice daily. Skin thickness, scratching behavior, and trans epidermal water loss (TEWL) were monitored continuously during the imiquimod application. Immediately after the final imiquimod application, ear skin and auricular lymph node (LN) were sampled for further analysis. Only a slight increase was observed in skin thickness in the citrinin exposure group; however, citrinin exposure significantly exacerbated hyperkeratinization and inflammatory cell infiltration in histological evaluation. TEWL, which is representative of cutaneous barrier function, was significantly increased by citrinin exposure. In terms of immune function, the number of immune cells in LN (T cells and dendritic cells) and gene expression of interleukin (IL)-17 in skin tissue were significantly increased by citrinin exposure. Direct interaction of dendritic cells (DCs) in citrinin-induced psoriasis development was further examined by proinflammatory cytokine determination in THP-1 cells and murine bone marrow derived DCs. IL-6 and/or tumor necrosis factor α were significantly increased by citrinin exposure. Taken together, our results imply that oral exposure to citrinin exacerbates the symptoms of a mouse model of psoriasis via direct activation of DCs.
Assuntos
Citrinina , Psoríase , Feminino , Animais , Camundongos , Imiquimode/toxicidade , Citrinina/toxicidade , Citrinina/metabolismo , Aminoquinolinas/toxicidade , Aminoquinolinas/metabolismo , Células Dendríticas , Psoríase/induzido quimicamente , Pele , Modelos Animais de Doenças , Camundongos Endogâmicos BALB CRESUMO
Inflammatory bowel disease (IBD) is a chronic inflammatory disease in the colon characterized by excessive activation of T cells. Glycosphingolipids (GSLs) are composed of lipid rafts in cellular membranes, and their content is linked to immune cell function. In the present study, we investigated the involvement of GSLs in IBD. Microarray data showed that in IBD patients, the expression of only UDP-glucose ceramide glucosyltransferase (UGCG) decreased among the GSLs synthases. Ad libitum access to dextran sulfate sodium (DSS) resulted in decreased UGCG and glucosylceramide (GlcCer) content in mesenteric lymph nodes and T cells from the spleen. Furthermore, the knockdown of Ugcg in T cells exacerbated the pathogenesis of colitis, which was accompanied by a decrease in Treg levels. Treatment with GlcCer nanoparticles prevented DSS-induced colitis. These results suggested that GlcCer in T cells is involved in the pathogenesis of IBD. Furthermore, GlcCer nanoparticles are a potential efficacious therapeutic target for IBD patients.
Assuntos
Glucosilceramidas/metabolismo , Glucosiltransferases/metabolismo , Doenças Inflamatórias Intestinais/patologia , Linfócitos T/metabolismo , Animais , Colite/induzido quimicamente , Colite/tratamento farmacológico , Colite/patologia , Sulfato de Dextrana/toxicidade , Modelos Animais de Doenças , Glucosilceramidas/administração & dosagem , Glucosilceramidas/genética , Glucosiltransferases/genética , Humanos , Doenças Inflamatórias Intestinais/metabolismo , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Nanopartículas/administração & dosagem , Nanopartículas/química , Linfócitos T/patologiaRESUMO
Deoxynivalenol (DON) contamination in food is a public health concern; however, the effect of DON exposure on immune disorders including allergies remains unclear. The aim of this study is to elucidate the effect of oral exposure to DON on pro-inflammatory and pro-pruritic responses in a mouse model of allergic dermatitis, which was generated by topical application of toluene-2,4-diisocyanate (TDI), a hapten that induces type-2 helper T cells. To evaluate acute exposure to DON, the mice were orally administered vehicle alone, 0.1 mg/kg DON, or 0.3 mg/kg DON 48, 24, and 1 h before the final challenge with TDI. To study subacute exposure, the mice were fed DON-contaminated rodent diet (0.3 ppm) during the experimental period. After the itch behavior and ear-swelling response were monitored, the serum, auricular lymph node, and skin tissue were collected for analyzing immunocyte differentiation, cytokine determination, and histological changes. Acute oral administration of DON significantly enhanced pro-inflammatory responses including ear-swelling response, immunocyte infiltration, and cytokine productions. Histological evaluation supported the occurrence of pro-inflammatory responses. In contrast, acute DON exposure only slightly increased itch behavior. Subacute oral exposure to DON significantly up-regulated the inflammatory responses, but showed almost no effect on pruritic response. In vitro evaluation in dendritic cells and keratinocytes indicated that DON pre-exposure induced a dose-dependent significant increase in cytokine production. Our results imply that both acute and subacute exposures to DON are associated with pro-inflammatory responses in cutaneous allergy.
Assuntos
Citocinas/metabolismo , Dermatite Alérgica de Contato/etiologia , Mediadores da Inflamação/metabolismo , Prurido/induzido quimicamente , Pele/efeitos dos fármacos , Células Th2/efeitos dos fármacos , Tricotecenos/toxicidade , Administração Oral , Animais , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Dermatite Alérgica de Contato/imunologia , Dermatite Alérgica de Contato/metabolismo , Dermatite Alérgica de Contato/patologia , Modelos Animais de Doenças , Feminino , Humanos , Queratinócitos/efeitos dos fármacos , Queratinócitos/imunologia , Queratinócitos/metabolismo , Camundongos Endogâmicos BALB C , Prurido/imunologia , Prurido/metabolismo , Prurido/patologia , Pele/imunologia , Pele/metabolismo , Pele/patologia , Células THP-1 , Células Th2/imunologia , Células Th2/metabolismo , Tolueno 2,4-Di-Isocianato , Tricotecenos/administração & dosagemRESUMO
We recently demonstrated that benzo[a]pyrene (BaP), the aryl hydrocarbon receptor (AhR) ligand, directly contributes to aggravation of cutaneous allergy in a mouse model of allergic dermatitis. The present study aimed to determine whether BaP-induced AhR activation results in development of airway inflammation. Initially, the potential for a direct relationship between BaP-induced AhR activation and airway inflammation was investigated in vivo, using a mouse model of type 2 helper T cell (Th2) hapten toluene-2,4-diisocyanate (TDI)-induced airway inflammation. Mice were orally administered BaP at 48, 24, and 4 h before the final allergen challenge. Oral administration of BaP showed a significant increase in lung inflammation and eosinophil infiltration. While expression of Th2 cytokines such as interleukin 4 (IL-4) and IL-13 was not affected by exposure to BaP, AhR activation significantly increased IL-33 expression. To confirm the in vivo results, in vitro experiments were performed using the human eosinophilic leukemia cell line (EOL-1), human bronchial epithelial cell line (BEAS-2B), and human lung adenocarcinoma epithelial cell line (A549). Results indicated that pre-treatment with BaP increased expression of IL-8 in house dust mite-activated EOL-1, BEAS-2B, and A549 cells. In addition, IL-33 levels in BEAS-2B cells were significantly increased after BaP exposure. Our findings indicated that BaP-induced AhR activation is involved in the pro-inflammatory response in respiratory allergy, and that this effect may be mediated by increased IL-33 expression and eosinophil infiltration.
Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/agonistas , Benzo(a)pireno/toxicidade , Quimiotaxia de Leucócito/efeitos dos fármacos , Eosinófilos/efeitos dos fármacos , Interleucina-33/metabolismo , Pulmão/efeitos dos fármacos , Pneumonia/induzido quimicamente , Receptores de Hidrocarboneto Arílico/agonistas , Células A549 , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Modelos Animais de Doenças , Eosinófilos/imunologia , Eosinófilos/metabolismo , Feminino , Humanos , Interleucina-8/metabolismo , Pulmão/imunologia , Pulmão/metabolismo , Camundongos Endogâmicos BALB C , Pneumonia/imunologia , Pneumonia/metabolismo , Receptores de Hidrocarboneto Arílico/metabolismo , Transdução de Sinais , Tolueno 2,4-Di-Isocianato , Regulação para CimaRESUMO
Our recent study has reported that estrogen receptors (ERs) are involved in several types of allergy development. This study aims to investigate the possible relationship between ER activation and development of imiquimod-induced psoriasis-like dermatitis. A mouse model of imiquimod-induced psoriasis-like dermatitis was generated by 5 days of topical application of 5% of imiquimod cream on the back of the ear and the shaved back skin of male BALB/c mice. From the second day of applying 5% imiquimod cream, either ERα selective agonist (propylpyrazoletriol [PPT] 2.5 mg/kg) or ERß selective agonist (diarylpropionitrile, DPN; 2.5 mg/kg) was administered orally for four consecutive days. Immediately after the final imiquimod cream application, scratching behavior was video monitored for 2 hours. The ear-swelling response was determined by comparing ear thickness before and after the final application of imiquimod cream. Twenty-four hours after the final imiquimod application, back skin tissue and auricular lymph nodes were isolated under isoflurane anesthesia. Oral administration of PPT significantly induced itch behavior and proinflammatory responses, including the levels of interleukin (IL)-17 and IL-22, whereas DPN treatment did not influence either pruritic or proinflammatory responses. In addition, IL-23 contribution by dendritic cells was identified using ER agonists on pretreated lipopolysaccharide (LPS)-stimulated murine bone marrow derived dendritic cells (BMDCs). PPT also significantly enhanced IL-23 secretion by LPS-stimulated BMDCs. Our findings indicate that the activation of ERα, but not ERß, is directly associated with inflammatory and pruritic responses in a mouse model of the imiquimod-induced psoriasis by enhancing the secretion of IL-23 by dendritic cells.
Assuntos
Células Dendríticas/efeitos dos fármacos , Receptor alfa de Estrogênio/metabolismo , Imiquimode/toxicidade , Inflamação/induzido quimicamente , Interleucina-23/metabolismo , Prurido/induzido quimicamente , Psoríase/induzido quimicamente , Psoríase/fisiopatologia , Animais , Humanos , Masculino , Camundongos , Modelos AnimaisRESUMO
We evaluated the real effects of pollutants through a multi-generation study. We tested whether short-term exposure (48 h) of successive (first and second) generations of Chironomus yoshimatsui neonates (<24-h-old) to two acetylcholinesterase inhibitor insecticides, pyraclofos, and pirimicarb, would change insecticide sensitivity and life-cycle parameters over four generations. Additionally, we tested whether acetylcholinesterase (AChE) activity levels would be associated with this sensitivity change. Sensitivities (48 h EC50 value, using immobility as the endpoint) in chironomids (<24-h-old) and insect life-cycle parameters (the number of larvae per egg mass and adult size) were investigated. Parental chironomids produced larvae that were less sensitive than those in the control group following the two 48 h pirimicarb exposure events, whereas exposure to pyraclofos did not affect sensitivity. The AChE activity in larvae with low sensitivity to pirimicarb was significantly higher than that in the control. Thus, increased AChE activity might be associated with low sensitivity. The life-cycle parameters in chironomids recovered from the effects of pyraclofos and pirimicarb suggested they could adapt to the insecticides by changing biomass allocation. Our study suggested potential chemical risks of insecticide stress and how aquatic organisms adapt to it.
Assuntos
Carbamatos/toxicidade , Chironomidae/efeitos dos fármacos , Inibidores da Colinesterase/toxicidade , Inseticidas/toxicidade , Organotiofosfatos/toxicidade , Pirimidinas/toxicidade , Adaptação Biológica/efeitos dos fármacos , Animais , Chironomidae/fisiologia , Ecotoxicologia/métodos , Larva/efeitos dos fármacos , Poluentes Químicos da Água/toxicidadeRESUMO
Neuromedin B is expressed in nociceptive and itch-sensitive dorsal root ganglia neurons, but its peripheral pruritogenic potential is not well described. The potential of neuromedin B as a pruritogen and pro-inflammatory peptide in the skin was tested in vivo in an acute model in mice and monkeys as well as an allergic dermatitis model in mice. To identify the underlying mechanisms in vitro real time PCR analysis for neuromedin B and its receptor expression in murine mast cells and dorsal root ganglia as well as functional calcium imaging in the ganglia was applied. Neuromedin B induces itch when injected intradermally, and the peripheral signal is likely transmitted through the activation of dorsal root ganglia. Thus, neuromedin B could be an interesting new therapeutic target for peripheral processing of itch at the level of sensory neurons.
Assuntos
Degranulação Celular , Mastócitos/fisiologia , Neurocinina B/análogos & derivados , Prurido/induzido quimicamente , Células Receptoras Sensoriais/fisiologia , Animais , Cálcio/metabolismo , Células Cultivadas , Dermatite Alérgica de Contato/etiologia , Feminino , Gânglios Espinais/citologia , Gânglios Espinais/metabolismo , Expressão Gênica , Indóis/farmacologia , Injeções Intradérmicas , Macaca mulatta , Masculino , Mastócitos/metabolismo , Camundongos , Neurocinina B/administração & dosagem , Piridinas/farmacologia , RNA Mensageiro/metabolismo , Receptores da Bombesina/antagonistas & inibidores , Receptores da Bombesina/genética , Receptores da Bombesina/metabolismo , Análise de Célula Única , Tolueno 2,4-Di-IsocianatoRESUMO
The aryl hydrocarbon receptor (AhR) is a ligand-dependent transcription factor that binds to various environmental chemicals and contributes to numerous toxicological effects. However, the direct effects of AhR on the development of allergic diseases are not fully understood. The main aim of this study was to elucidate the action of AhR in the development of cutaneous allergies. Initially, the potential for a direct relationship between AhR and the immune cells was investigated in vitro, using murine bone marrow-derived dendritic cells, human epidermal keratinocytes, and the mixed leukocyte reaction assay. Benzo[a]pyrene (BaP) and 6-formylindolo[3,2-b]carbazole were used as selective ligands for the AhR. Pretreatment with BaP and/or 6-formylindolo[3,2-b]carbazole significantly induced cytokine release by activated keratinocytes and T-cell proliferation, whereas interleukin-12 production in bone marrow-derived dendritic cells was reduced by AhR activation. To confirm the in vitro results, in vivo experiments were also performed in T-helper (Th)2-type hapten toluene-2,4-diisocyanate- and Th1-type hapten dinitrochlorobenzene-induced mouse models of allergic dermatitis. Mice were orally administered BaP at 48, 24 and 4 hours before the final allergen challenge. In the Th2 model, ear-swelling response and scratching behavior were promoted by BaP exposure, which supported the observed significant increases in local cytokine secretion. The infiltration of helper T cells, B cells and dendritic cells into the auricular lymph node was significantly enhanced by BaP administration, although Th1-type immune responses were not influenced by AhR activation. Our findings demonstrate that AhR activation directly activates keratinocytes and T cells, which leads to the exacerbation of Th2-type cutaneous allergy.
Assuntos
Benzo(a)pireno/toxicidade , Células Dendríticas/efeitos dos fármacos , Dermatite Alérgica de Contato/imunologia , Queratinócitos/efeitos dos fármacos , Receptores de Hidrocarboneto Arílico/metabolismo , Células Th2/imunologia , Animais , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Citocinas/metabolismo , Células Dendríticas/imunologia , Dermatite Alérgica de Contato/etiologia , Dermatite Alérgica de Contato/metabolismo , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Feminino , Queratinócitos/imunologia , Teste de Cultura Mista de Linfócitos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Células Th2/efeitos dos fármacosRESUMO
Allergic diseases are driven by activation of mast cells and release of mediators in response to IgE-directed antigens. However, there are no drugs currently available that can specifically down-regulate mast cell function in vivo when chronically administered. Here, we describe an innovative approach for targeting mast cells in vitro and in vivo using antisense oligonucleotide-mediated exon skipping of the ß-subunit of the high-affinity IgE receptor (FcεRIß) to eliminate surface high-affinity IgE receptor (FcεRI) expression and function, rendering mast cells unresponsive to IgE-mediated activation. As FcεRIß expression is restricted to mast cells and basophils, this approach would selectively target these cell types. Given the success of exon skipping in clinical trials to treat genetic diseases such as Duchenne muscular dystrophy, we propose that exon skipping of FcεRIß is a potential approach for mast cell-specific treatment of allergic diseases.
Assuntos
Degranulação Celular/efeitos dos fármacos , Dermatite Alérgica de Contato/terapia , Mastócitos/efeitos dos fármacos , Oligonucleotídeos Antissenso/uso terapêutico , Splicing de RNA/efeitos dos fármacos , Receptores de IgE/metabolismo , Animais , Cálcio/metabolismo , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Citocinas/biossíntese , Modelos Animais de Doenças , Feminino , Humanos , Mastócitos/metabolismo , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Oligonucleotídeos Antissenso/farmacologia , Anafilaxia Cutânea Passiva/genética , Receptores de IgE/genéticaRESUMO
It has been reported that endogenous or exogenous estrogens can affect the immune system, resulting in immune disorders; however, their direct involvement in such conditions remains to be demonstrated. The purpose of this study was to investigate whether estrogen receptors (ER) are directly implicated in pro-pruritic and pro-inflammatory reactions in cutaneous allergy. Initially, enhancement of the pro-inflammatory response by several ER agonists [methoxychlor (MXC), ß-estradiol (E2), propylpyrazoletriol (PPT; an ERα agonist), and diarylpropionitrile (DPN; an ERß agonist)] was examined in vivo using a male BALB/c mouse model of allergic dermatitis induced by toluene-2,4-diisocyanate administration. The ear swelling response, itch response, and local cytokine secretion were measured. Subsequently, the mechanism underlying the development of such allergic reactions was analyzed in vitro using human epidermal keratinocytes, murine bone marrow-derived dendritic cells (mBMDCs), and the mixed leucocyte reaction assay. Activated cells were exposed to each ER agonist for 24â¯h, and cytokine secretion and cell proliferation were measured. Our in vivo experiments indicated significant upregulation of pro-inflammatory and pro-pruritic responses in the E2-, MXC-, and PPT-treated groups compared to the control group; however, no change was observed in the DPN-treated group. Levels of cytokines expressed by keratinocytes, such as TSLP and IL-33, were particularly increased by exposure to E2, MXC, or PPT. These in vivo results were confirmed in vitro in keratinocytes, but not mBMDCs or T cells. Our findings imply that ERα is involved in pro-inflammatory and pro-pruritic responses in cutaneous allergy through activation of keratinocytes.
Assuntos
Dermatite Alérgica de Contato/tratamento farmacológico , Moduladores de Receptor Estrogênico/uso terapêutico , Receptor alfa de Estrogênio/efeitos dos fármacos , Inflamação/tratamento farmacológico , Prurido/tratamento farmacológico , Animais , Proliferação de Células/efeitos dos fármacos , Citocinas/metabolismo , Células Dendríticas , Dermatite Alérgica de Contato/patologia , Orelha Externa/patologia , Feminino , Humanos , Inflamação/patologia , Queratinócitos/metabolismo , Teste de Cultura Mista de Linfócitos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Prurido/patologiaRESUMO
The prevalence of allergic skin disorders has increased rapidly, and development of therapeutic agents to alleviate the symptoms are still needed. In this study, we orally or topically administered the Janus kinase (JAK) inhibitors, tofacitinib and oclacitinib, in a mouse model of dermatitis, and compared the efficacy to reduce the itch and inflammatory response. In vitro effects of JAK inhibitors on bone marrow-derived dendritic cells (BMDCs) were analyzed. For the allergic dermatitis model, female BALB/c mice were sensitized and challenged with toluene-2,4-diisocyanate (TDI). Each JAK inhibitor was orally or topically applied 30 minutes before and 4 hours after TDI challenge. After scratching bouts and ear thickness were measured, cytokines were determined in challenged skin and the cells of the draining lymph node were analyzed by means of flow cytometry. In vitro, both JAK inhibitors significantly inhibited cytokine production, migration, and maturation of BMDCs. Mice treated orally with JAK inhibitors showed a significant decrease in scratching behavior; however, ear thickness was not significantly reduced. In contrast, both scratching behavior and ear thickness in the topical treatment group were significantly reduced compared with the vehicle treatment group. However, cytokine production was differentially regulated by the JAK inhibitors, with some cytokines being significantly decreased and some being significantly increased. In conclusion, oral treatment with JAK inhibitors reduced itch behavior dramatically but had only little effect on the inflammatory response, whereas topical treatment improved both itch and inflammatory response. Although the JAK-inhibitory profile differs between both JAK inhibitors in vitro as well as in vivo, the effects have been comparable.
Assuntos
Anti-Inflamatórios/administração & dosagem , Antipruriginosos/administração & dosagem , Dermatite Alérgica de Contato/tratamento farmacológico , Janus Quinases/antagonistas & inibidores , Piperidinas/administração & dosagem , Pirimidinas/administração & dosagem , Pirróis/administração & dosagem , Sulfonamidas/administração & dosagem , Administração Oral , Administração Tópica , Animais , Citocinas/metabolismo , Dermatite Alérgica de Contato/metabolismo , Modelos Animais de Doenças , Feminino , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Linfonodos/efeitos dos fármacos , Linfonodos/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Prurido/tratamento farmacológico , Prurido/metabolismo , Pele/efeitos dos fármacos , Pele/metabolismoRESUMO
Abstract Humans are exposed daily to multiple environmental chemicals in the atmosphere, in food, and in commercial products. Therefore, hazard identification and risk management must account for exposure to chemical mixtures. The objective of the study reported here was to investigate the effects of combinations of three well-known environmental immunotoxic chemicals - methoxychlor (MXC), an organochlorine compound; parathion (PARA), an organophosphate compound; and piperonyl butoxide (PBO), an agricultural insecticide synergist - by using a mouse model of ovalbumin (OVA)-induced allergic airway inflammation. Four-week-old Balb/c mice were exposed orally to either one or two of the environmental immunotoxic chemicals for five consecutive days, prior to intraperitoneal sensitization with OVA and an inhalation challenge. We assessed IgE levels in serum, B-cell counts, and cytokine production in hilar lymph nodes, and differential cell counts and levels of related chemokines in bronchoalveolar lavage fluid (BALF). Mice treated with MXC + PARA or PBO + MXC showed marked increases in serum IgE, IgE-positive B-cells and cytokines in lymph nodes, and differential cell counts and related chemokines in BALF compared with mice that received the vehicle control or the corresponding individual test substances. These results suggest that simultaneous exposure to multiple environmental chemicals aggravates allergic airway inflammation more than exposure to individual chemicals. It is expected that the results of this study will help others in their evaluation of immunotoxic combinational effects when conducting assessments of the safety of environmental/occupational chemicals.
Assuntos
Poluentes Ambientais/toxicidade , Imunossupressores/toxicidade , Hipersensibilidade Respiratória/induzido quimicamente , Administração Oral , Animais , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Citocinas/imunologia , Sinergismo Farmacológico , Poluentes Ambientais/química , Desenho de Equipamento , Feminino , Citometria de Fluxo , Imunoglobulina E/sangue , Imunossupressores/química , Exposição por Inalação , Injeções Intraperitoneais , Linfonodos/citologia , Linfonodos/efeitos dos fármacos , Linfonodos/imunologia , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia , Hipersensibilidade Respiratória/imunologia , Fatores de Tempo , Testes de Toxicidade/instrumentação , Testes de Toxicidade/métodosRESUMO
In dogs, Porphyromonas gulae is a major periodontal pathogen with 41-kDa proteins polymerizing to form a filamentous structure called fimbriae or pili, termed FimA. FimA is classified into three genotypes: A, B, and C, and there are combinations of types A, B, C, A/B, A/C, B/C, and A/B/C. Periodontal disease is the most common oral disease in small dogs, but the periodontal disease status and P. gulae colonization at each dog age and breed remain unclear. In this study, we stratified 665 small dogs and analyzed the periodontal status and distribution of P. gulae with each FimA genotype. Dogs with periodontal disease and FimA genotype tended to increase with age. The dogs with at least one FimA genotype had significantly more severe periodontal disease compared with P. gulae-negative dogs (P < 0.01). Additionally, periodontal status was significantly associated with specific FimA genotype distribution in Toy Poodles and Chihuahuas (P < 0.05), whereas there was no such association in Dachshunds. These results suggest that the onset of periodontal disease and P. gulae colonization are related and progress with age. The relationship between periodontal disease and FimA genotype may differ depending on the dog breeds.