Selective Inhibition of β-Catenin/Co-Activator Cyclic AMP Response Element-Binding Protein-Dependent Signaling Prevents the Emergence of Hapten-Induced Atopic Dermatitis-Like Dermatitis

BACKGROUND: The canonical Wnt/β-catenin signaling pathway is a fundamental regulatory system involved in various biological events. ICG-001 selectively blocks the interaction of β-catenin with its transcriptional co-activator cyclic AMP response element-binding protein (CBP). Recent studies have provided convincing evidence of the inhibitory effects of ICG-001 on Wnt-driven disease models, such as organ fibrosis, cancer, acute lymphoblastic leukemia, and asthma. However, the effects of ICG-001 in atopic dermatitis (AD) have not been investigated. OBJECTIVE: To investigate whether β-catenin/CBP-dependent signaling was contributed in the pathogenesis of AD and ICG-001 could be a therapeutic agent for AD. METHODS: We examined the effects of ICG-001 in an AD-like murine model generated by repeated topical application of the hapten, oxazolone (Ox). ICG-001 or vehicle alone was injected intraperitoneally every day during the development of AD-like dermatitis arising from once-daily Ox treatment. RESULTS: Ox-induced AD-like dermatitis characterized by increases in transepidermal water loss, epidermal thickness, dermal thickness accompanied by increased myofibroblast and mast cell counts, and serum levels of thymic stromal lymphopoietin and thymus and activation-regulated chemokine, and decreases in stratum corneum hydration, were virtually normalized by the treatment with ICG-001. Elevated serum levels of periostin tended to be downregulated, without statistical significance. CONCLUSION: These results suggest that β-catenin/CBP-dependent signaling might be involved in the pathogenesis of AD and could be a therapeutic target.

Dioscorea japonica Thunb. Ethanolic Extract Attenuated Oxazolone-Induced Atopic Dermatitis-like Skin Lesions in BALB/c Mice

The rhizomes of Dioscorea japonica Thunb. are widely consumed as food and also used to treat diabetes and polyuria in Korea. This study was undertaken to study the anti-atopic dermatitis effects of a 95% ethanolic extract (DJE) of D. japonica in an oxazolone-stimulated murine model of atopic dermatitis (AD). The therapeutic effects of DJE on AD-like skin lesions were assessed on both ears. DJE (1%) or dexamethasone (0.5%; the positive control) were applied to skin lesions for three weeks. Serum levels of IgE and IL-4 were assessed by ELISA (enzyme-linked immunosorbent assay). Histopathological examinations were performed by hematoxylin and eosin (H&E) and toluidine blue staining and revealed DJE significantly reduced dermal thickness and inflammatory cell infiltration when applied to oxazolone-treated ear skin. DJE-treated AD mice also showed lower serum levels of IgE and IL-4 than oxazolone-stimulated controls. Our findings demonstrate DJE might be a useful safe, topical agent for the treatment of atopic diseases.

Potent anti-inflammatory activity of tetramethylpyrazine is mediated through suppression of NF-k

The purpose of the current study was to evaluate the anti-inflammatory activity of tetramethlpyrazine on oxazolone-induced colitis mice. Spleen mononuclear cells [SMC], lamina propria mononuclear cells [LPMC] and peripheral blood mononuclear cells [PBMC] were isolated from oxazolone-induced colitis and normal mice. The colitis cells treated by oxazolone were randomly divided into model, low dose, middle dose and high dose groups treated with 0, 0.5, 1.0 and 2.0 g/L tetramethlpyrazine, respectively. The apoptotic rate of SMC and LPMC in the oxazolone-induced group was lower than that in the normal group. Compared with model group, apoptotic rate of SMC was significantly increased in the high dose group, while the apoptotic rate of LPMC in the middle dose group was increased. Compared with SMC, LPMC and PBMC of normal group, the mRNA level of nuclear factor kappa B [NF-kB], transcription factor-activated protein-1 [AP-1] and nuclear factor of activated T cells [NF-AT] were higher in model group. Tetramethylpyrazine inhibited the increase of NF-kB, AP-1 and NF-AT mRNA induced by oxazolone. For SMC, LPMC and PBMC there was significant difference in the mRNA level of AP-1 among the three different doses of tetramethylpyrazine treated groups. However, no significant difference was observed in the mRNA levels of NF-AT and NF-kB between normal and middle groups. Tetramethylpyrazine promoted the apoptotic rate of SMC and LPMC in-vitro, and suppressed the expression of transcription factors in SMC, LPMC and PBMC isolated from oxazolone-induced colitis mice. The study provides a novel insight into the mechanism behind the effect of etramethylpyrazine on colitis

Ethanol Extract of Peanut Sprout Exhibits a Potent Anti-Inflammatory Activity in Both an Oxazolone-Induced Contact Dermatitis Mouse Model and Compound 48/80-Treated HaCaT Cells

BACKGROUND: We developed an ethanol extract of peanut sprouts (EPS), a peanut sprout-derived natural product, which contains a high level of trans-resveratrol (176.75 microg/ml) and was shown to have potent antioxidant activity. OBJECTIVE: We evaluated the potential anti-inflammatory activity of EPS by measuring its antioxidant potential in skin. METHODS: The anti-inflammatory activity of EPS was tested using two models of skin inflammation: oxazolone (OX)-induced contact dermatitis in mice and compound 48/80-treated HaCaT cells. As biomarkers of skin inflammation, cyclooxygenase-2 (COX-2) and nerve growth factor (NGF) levels were measured. RESULTS: OX-induced contact dermatitis was suppressed markedly in mice that were treated with an ointment containing 5% EPS as evidenced by a decrease in the extent of scaling and thickening (p<0.05) and supported by a histological study. COX-2 (messenger RNA [mRNA] and protein) and NGF (mRNA) levels, which were upregulated in the skin of OX-treated mice, were suppressed markedly in the skin of OX+EPS-treated mice. Consistent with this, compound 48/80-induced expression of COX-2 (mRNA and protein) and NGF (mRNA) in HaCaT cells were suppressed by EPS treatment in a dose-dependent manner. As an inhibitor of NF-kappaB, IkappaB protein levels were dose-dependently upregulated by EPS. Fluorescence-activated cell sorting (FACS) analysis revealed that EPS scavenged compound 48/80-induced reactive oxygen species (ROS) in HaCaT cells. CONCLUSION: EPS exerts a potent anti-inflammatory activity via its anti-oxidant activity in both mouse skin and compound 48/80-treated HaCaT cells in vitro. Compound 48/80-treated HaCaT cells are a useful new in vitro model of skin inflammation.

A Review on Chemical-Induced Inflammatory Bowel Disease Models in Rodents

Ulcerative colitis and Crohn's disease are a set of chronic, idiopathic, immunological and relapsing inflammatory disorders of the gastrointestinal tract referred to as inflammatory bowel disorder (IBD). Although the etiological factors involved in the perpetuation of IBD remain uncertain, development of various animal models provides new insights to unveil the onset and the progression of IBD. Various chemical-induced colitis models are widely used on laboratory scale. Furthermore, these models closely mimic morphological, histopathological and symptomatical features of human IBD. Among the chemical-induced colitis models, trinitrobenzene sulfonic acid (TNBS)-induced colitis, oxazolone induced-colitis and dextran sulphate sodium (DSS)-induced colitis models are most widely used. TNBS elicits Th-1 driven immune response, whereas oxazolone predominantly exhibits immune response of Th-2 phenotype. DSS-induced colitis model also induces changes in Th-1/Th-2 cytokine profile. The present review discusses the methodology and rationale of using various chemical-induced colitis models for evaluating the pathogenesis of IBD.

The Effect of Adipose-Derived Stem Cell-Cultured Media on Oxazolone Treated Atopic Dermatitis-Like Murine Model

BACKGROUND: A stem cell is an undifferentiated cell that has the potential for self-renewal and differentiation. Adipose-derived stem cells (ADSCs) have advantages in accessibility and abundance compared to other kinds of stem cells and produce many growth factors and hormones. OBJECTIVE: We investigated whether ADSC cultured media could be used as a therapy for atopic dermatitis. METHODS: ADSC cultured media was topically applied twice daily for 5 days to oxazolone-treated atopic dermatitis-like hairless mice. RESULTS: Topical application of ADSC cultured media improved the epidermal permeability barrier and keratinocyte differentiation, and restored the predominant Th2 phenotype when compared to vehicle. ADSC cultured media-treated epidermis also showed an increase in the expression of antimicrobial peptides cathelin-related antimicrobial peptide, mouse beta-defensein 3. CONCLUSION: Topical ADSC cultured media could be useful in the treatment of atopic dermatitis.

Paeoniflorin increases beta-defensin expression and attenuates lesion in the colonic mucosa from mice with oxazolone-induced colitis / 药学学报

Previous studies have demonstrated that the Chinese medicine paeoniflorin, derived from the Ranunculaceae plant peony, peony, purple peony root, was able to have anti-inflammatory, anti-ulcer, anti-hypersusceptibility and anti-oxidation activity. In order to elucidate the pesticide effect and the mechanisms by which paeoniflorin exerts its effect of anti-inflammation and immunoregulation on oxazolone-induced colitic mice, disease activity index (DAI) and histological grading of colitis (HGC) were evaluated in animal model. Moreover, the expressions of HBD-2, IL-6 and IL-10 of mice with experimental colitis were observed with immunohistochemistry and RT-PCR in this study. Results showed that DAI and HGC of oxazolone control group was significantly higher than that of normal control group, and that paeoniflorin groups and 5-ASA group, compared with oxazolone control group, could alleviate the symptoms and histological damages of colitic mice (P < 0.05, P < 0.01). The expression of HBD-2 and IL-6 cytokine on the colon of colitic mice was higher than that of normal control, paeoniflorin and 5-ASA groups (P < 0.05, P < 0.01), but the expression of IL-10 is lower than that of normal control, paeoniflorin and 5-ASA groups (P < 0.05, P < 0.01). The positive correlations were demonstrated between the expression of (HBD-2 and IL-6) and DAI (Pearson r = 0.728, Pearson r = 0.758, P < 0.01, respectively), (HBD-2 and IL-6) and HGC (Pearson r = 0.819, Pearson r = 0.825, P < 0.01, respectively), whereas, the negative correlations were demonstrated between the expression of IL-10 and DAI (Pearson r = -0.789, P < 0.01), IL-10 and HGC (Pearson r = -0.725, P < 0.01). It can be concluded that to some extent paeoniflorin effectively alleviate the symptoms of oxazolone-induced colitis through regulating the expression of HBD-2, IL-6 and IL-10.

Overexpression of CIITA in T Cells Aggravates Th2-Mediated Colitis in Mice

The MHC class II transactivator (CIITA) is the master transcriptional regulator of genes involved in MHC class II restricted antigen presentation. Previously we suggested another role of CIITA in Th1/Th2 balance by demonstrating that forced expression of CIITA in murine T cells repressed Th1 immunity both in vitro and in vivo. However, the results were contradictory to the report that CIITA functioned to suppress the production of Th2 cytokine by CD4+T cells in CIITA deficient mice. In this study, we investigated the influence of constitutive expression of CIITA in T cells on Th2 immune response in vivo using murine experimental colitis model. In the dextran sodium sulfate-induced acute colitis, a disease involving innate immunity, CIITA transgenic mice and wild type control mice showed similar progression of the disease. However, the development of oxazolone-induced colitis, a colitis mediated by predominantly Th2 immune response, was aggravated in CIITA-transgenic mice. And, CD4+T cells from the mesenteric lymph node of CIITA-transgenic mice treated with oxazolone exhibited a high level of IL-4 secretion. Together, these data demonstrate that constitutive expression of CIITA in T cells skews immune response to Th2, resulting in aggravation of Th2-mediated colitis in vivo.

Studies on the binary complexes of Ni [II], Cu [II] and Pt [IV] with oxazolidine and imidazolidine derivatives

Some oxazolidine and imidazolidine derivatives were synthesized to investigate their complexing behavior with Cu [II], Ni [II] and Pt [IV] ions. Ni [II], Cu [II] and Pt [IV] complexes with 5-imino-3- [p-fluorophenyl]-2-methyloxazolidine-4-thione [Pfox] and with 5-imino- 1-phenyl-3[p-fluorophenyl]-2-imidazolidine-4-thione [pfim] were characterized by elemental analysis, TGA, IR, electronic absorption spectra, molar conductance, and X-ray diffraction studies. Complexation equilibria and the stabilities of these binary complexes obtained in solution were studied potentiometrically and spectrophotometrically in a 1:1 molar ratio in 40% ethanol-water mixture at the constant ionic strength [I = 0.1 M NaClO4]

The Effect of High Dose UVA - 1 and UVA - 2 Irradiation on the Expression of Surface Markers of Epidermal Langerhans Cells and Induction of contact Hypersensitivity in Mice Skin / 대한피부과학회지

BACKGROUND: It is knovn that Langerhans cells are damaged fuctionally and morphologically by UV irradiation. Recently, high-dose UVA-1 therapy (340-400nm) was introduced as an effective treatment of severe exacerbated atopic dermatitis. However, the effect of UVA-1 therapy on surface markers and function of epidermal Langerhans cells are still unclear. OBJECTIVE: To determine whether a high dose UVA-1 irradiation affects cutaneous immune system, the effect of UVA-1 on the expression of ATPase and Ia antigen of mouse epidermal Langerhans cells and induction of contact hypersensitivity in mice skin were investigated and were compared to those of UVA-2. METHODS: Balb/c mice were irradiated with 150J/cm and 300J/cm of UVA-1 and UVA-2 in a single dose at one time or 3 fractionated doses for 3 days. The number of Langerhans cells was evaluated using ATPase and immunoperoxidase-stained epidermal sheets. Balb/c mice were irradiated with same manner after induction of contact hypersensiyity by applying 0.5% oxazolone solution and the influence of UV irradiation was evaluated by measuring the ear swelling of mice. RESULTS: 1. The expression of surface markers of Langerhans cells was not affected by 150J/cm and fractionated 300J/cm of UVA-1. However, single irradiation of 300J/cm of UVA-1 reduced signifi-cantly the expression of surface markers. The irradiation of UVA-2 induced more prominent reduction of the expression of surface markers compared to UVA-l. 2. Although the induction of contact hypersensitity was not inhibited in groups irradiated by single or fractionated 150J/cm of UVA-1, it was inhibited in groups irradiated with 300J/cm of UVA-1. The inhibition of contact hypersensitivity induction by UVA-2 irradiation was also more prominent than that by UVA-1. CONCLUSION: These results suggest that epidermal Langerhans cells could be damaged by high doses of UVA-1 and the damage of Langerhans cells by UVA-1 is weaker than that by UVA-2.

Estudo do tráfego linfocitário e sua modulação em camundongos, in vivo

Uma compreensão aprofundada dos mecanismos envolvidos na migração linfocitária e na sua modulação, permitiria que essas células fossem direcionadas artificialmente. Isto poderia impedir sua entrada e acúmulo em sítios indesejáveis (como por exemplo, órgãos alvo em doenças auto-imunes ou órgãos transplantados) ou, alternativamente, seria possível facilitar sua em uma situação em que o acúmulo de células efetoras funcionais fosse mandatório, como seria o caso de tumores. No presente trabalho este problema foi abordado estabelecendo-se, inicialmente, um sistema de estudo para padronização da distribuição do trêfego linfocitário, em camundongos normais, e em seguida procurou-se uma possibilidade de modular este tráfego. Foram utilizadas células linfóides marcadas com51 Cr e injetadas por via endovenosa. Duas substâncias, o antiinflamatório não esteróide Indometacina (INDO) e o agente imunossupressor Ciclosporina A (CS-A) foram utilizadas com o objetivo de verificar se seria possível modular esta distribuição linfocitária normal ou em situações inflamatórias desencadeadas por uma reação de sensibilidade de contato e por indução de tumor. CS-A foi capaz de alterar a migração linfocitária, em situações fisiológicas, para o intestino e timo enquanto INDO apenas induziu uma inibição da migração de linfoblastos periféricos para o timo. CS-A aumentou a permeabilidade do timo a linfoblastos periféricos e induziu um maior acúmulo de linfoblastos mesenféricos no intestino. Por outro lado, o tratamento de infiltrados inflamatórios com CS-A produziu efeitos paradoxais. CS-A inibiu o acúmulo de linfoblastos em tumores em pleno crescimento, mas só foi capaz de inibir parcialmente e tardiamente a entrada de células em lesões pelo sesibilizante de contato Oxazolona (OXA). Nas mesmas condições, INDO tem efeito comparável ao de CS-A, nas lesões induzidas por OXA. O tratamento deste infiltrado inflamtorio com ambas, CS-A e INDO não modificaram o efeito exercido por nenhuma delas, considerado isoladamente. O tratamento experimental com radiação, por si mesmo, induziu alterações na migração de células para o baço, LNM, intestino, pele, pulmão e timo. Tal tratamento levou ao acúmulo de linfoblastos no intestino e timo e, surpreendentemente, CS-A produziu um aumento considerável na ligração linfocitária para o intestino e timo de animais irradiados, mostrando um efeito sinergístico, sugerindo que o acúmulo dessas células nos vários modelos resulta de mecanismos diversos e que o uso de CSA não pode ser considerado de maneira exclusiva em inflamatórios de risco de linfócitos.

A better understanding of the mechanism involved on lymphocyte migration and modulation might allow these cells to be artificially directed. This could hinder the entrance and accumulation of these cells at inconvenient sites (target organs in autoimmune diseases or transplanted organs, for example) or alternatively, it woud be possible to facilitade their entrance in situations in which the accumulation of effector functional cells were mandatory as it would be case for tumors. In the present work this problem was dealt with, by initially establishing a system 1· of study to standardize the distribution of the lymphocyte traffic in normal mice and then ; it has been searched a manner of modulating such traffic. Lymphoid cells labeled with 51Cr and intravenously injected have been utilized. Two substances, the anti-inflammatory non steroid - Indometacin (INDO) and the immunosupressor agent - Cyclosporin A (CS-A), were utilized in order to verify the posibility of modulating lymphocyte distribution at inflammatory situations induced by several stimuli. CS-A has been able to alter the lymphocyte migration, in physiological situations, towards gut and thymus whereas INDO only induced an inhibition of the peripheric lymphoblasts migration towards the thymus. CS-A increasing thymus permeability to peripheric Iymphoblasts and induced a higher accumulation of mesenteric lymphoblasts in the gut. On the other hand, treatment of inflammatory infiltrates with CS-A produced paradoxical effects. CS-A inhibited the accumulation of lymphocytes in developing tumors but it was only able to inhibit the entrance of cells in lesions induced by Oxazolone (OXA), partially and at late stage. In the same conditions, INDO has comparable effect as CS-A, in lesions induced by OXA. The treatment of this inflammatory infiltrate with both CS-A and INDO did not modify the effect exerted by either of them, considered isolately. The experimental treatment with radiation, induced, on its own, modifications in the migration of cells towards spleen, mesenteric lymph nodes, gut, lungs and thymus. Such treatment led to the accumulation oflymphoblasts in the gut and surprisingly, CS-A promoted a considerable increase of the lymphocyte migration towards the gut and the thymus of irradiated animals, suggesting that such cell accumulation in the several models results from diverse mechanisms and that the use of CS-A canot be considered in an exclusive way on infiltrates rich with lymphocytes.

Synthesis and reactions of 2,2'- [m-Phenylene] bis [[4-arylidene-5 [4H] -oxazolones]

2'-[M-phenylene] bis [4-aryliaene-5 [4H]-oxazolones-Va-d have been synthesized. The hydrolysis, alcoholysis, ammonolysis, aminolysis as well as hydrazinolysis of these oxazolones have been studied. When the products of alcoholysis, aminonolysis and aminolysis were heated above their melting points they have eliminated 2 molecules of alcohol, ammonia and amine respectively yielding the starting oxazolones. Also it was noted, that the alcoholysis may be catalyzed by sodium azide. The spectroscopic data by the newly synthesized compounds are discussed. Some of the newly synthesized compounds were tested to determine their dyeing and gmepnti properties

Modulaçåo da resposta imune induzida por ciclofosfamida em Leishmaniose expeimental de Hamster / Effect of Cyclophosphamide in the modulation of immune response in the experimental Leishmaniose of the Hamster

Estudou-se a resposta imune da leishmaniose visceral em hamsteres inoculados com (10) x (7) amastigotas da Leishmania sp. cepa 70 por via intraperitonial. As alteraçöes imunológicas encontradas foram: produçåo de anticorpos anti-leishmânia, aumento da densidade linfocitária na zona B-dependente e reaçåo de hipersensibilidade tardia específica negativa. A modulaçåo da resposta imune com o uso da ciclofosfamida mostrou uma diminuiçåo no nível de anticorpos anti-leishmânia, depleçåo linfocitária da zona B-dependente, aparecimento da reaçåo de leishmania e sugeriu a existência de célula reguladora da reaçåo de hipersensibilidade tardia, sensível à ciclofosfamida, em hamster

Pattern of Oral Desensitization in DNCB Presensitized Guinea Pigs / 대한피부과학회지

The pattern of DNCB (2, 4-dinitrochlorobenzene) oral desensitization and its antigenic specificity were investigated in guinea pigs. In search of antigenic specificity of DNCB oral desensitization, animals were fed oxazolone (4-ethoxymethylene-2- phenyl-oxazol-5-one) in a DNCB presensitized group, and conversely, DNCB fed in an oxazolone presensitized group. For the study of the pattern of oral desensitization, guinea pigs were initially sensitized to DNCB and followed by feeding of DNCB for 6 days, and challenged on the 13th, 21st, 29th, 37th and 45th days after sensitization. Oxazolone feeding in DNCB presensitized guinea pigs had no effect on the development of the fully responsive DNCB contact sensitivity (72. 85g), and DNCB feeding in oxazolone pre-primed animal had no effect on the development of oxazolone contact sensitivity (79. 37g). On the contrary, oxazolone feeding in DNCB preprimed guinea pigs and DNCB feeding in oxazolone pre-primed resulted in, respectively, oxazolone and DNCB oral tolerance (44.44% p < 0.01 & 42.06%p<0.01). The effect of desensitization appeared even one day before the completion of 6 days' feeding and the efficacy lasted about 30 days by the natural waning of contact sensitivity.