Topical Application of Eupatilin Ameliorates Atopic Dermatitis-Like Skin Lesions in NC/Nga Mice

BACKGROUND: Atopic dermatitis (AD) is an inflammatory skin disorder with severe pruritus. Despite advancements in medicine, therapeutic treatments for AD are still limited. Eupatilin (5,7-dihydroxy-30,40,6-trimethoxyflavone) is one of the lipophilic flavonoids from Artemisia umbelliformis Lam. and Artemisia genipi Weber. OBJECTIVE: Although it has been reported to act a role in improving inflammation, its action on AD is uncertain. In this study, we examined the role of eupatilin on AD-like skin lesions in NC/Nga mice. METHODS: 2,4-dinitrochlorobenzene was repeatedly applied to the ear of NC/Nga mice to produce AD-like skin lesions. Eupatilin (1%, once a day for 5 consecutive days/week) was applied topically for four weeks for the evaluation of its therapeutic effects. RESULTS: 1% eupatilin cream significantly reduced the clinical severity score of AD-like lesions, compared to the vehicle (p<0.005). A histopathological analysis revealed that 1% eupatilin cream significantly decreased the mast cell infiltration as well as inflammatory cell infiltration, compared to the vehicle (p<0.005). We showed that 1% eupatilin cream significantly reduced the expression of thymic stromal lymphopoietin, tumor necrosis factor-α, interleukin-4, and interleukin-19, but not interferon-γ, compared to the vehicle (p<0.005). CONCLUSION: Considering the therapeutic reaction of eupatilin on AD-like lesions as in this study, the substance has a promising to be an adjuvant topical agent for the control of AD.

Topical Application of Eupatilin Ameliorates Atopic Dermatitis-Like Skin Lesions in NC/Nga Mice

BACKGROUND: Atopic dermatitis (AD) is an inflammatory skin disorder with severe pruritus. Despite advancements in medicine, therapeutic treatments for AD are still limited. Eupatilin (5,7-dihydroxy-30,40,6-trimethoxyflavone) is one of the lipophilic flavonoids from Artemisia umbelliformis Lam. and Artemisia genipi Weber. OBJECTIVE: Although it has been reported to act a role in improving inflammation, its action on AD is uncertain. In this study, we examined the role of eupatilin on AD-like skin lesions in NC/Nga mice. METHODS: 2,4-dinitrochlorobenzene was repeatedly applied to the ear of NC/Nga mice to produce AD-like skin lesions. Eupatilin (1%, once a day for 5 consecutive days/week) was applied topically for four weeks for the evaluation of its therapeutic effects. RESULTS: 1% eupatilin cream significantly reduced the clinical severity score of AD-like lesions, compared to the vehicle (p<0.005). A histopathological analysis revealed that 1% eupatilin cream significantly decreased the mast cell infiltration as well as inflammatory cell infiltration, compared to the vehicle (p<0.005). We showed that 1% eupatilin cream significantly reduced the expression of thymic stromal lymphopoietin, tumor necrosis factor-α, interleukin-4, and interleukin-19, but not interferon-γ, compared to the vehicle (p<0.005). CONCLUSION: Considering the therapeutic reaction of eupatilin on AD-like lesions as in this study, the substance has a promising to be an adjuvant topical agent for the control of AD.

Rifampicin Alleviates Atopic Dermatitis-Like Response in vivo and in vitro

Atopic dermatitis (AD) is a common inflammatory skin disorder mediated by inflammatory cells, such as macrophages and mast cells. Rifampicin is mainly used for the treatment of tuberculosis. Recently, it was reported that rifampicin has anti-inflammatory and immune-suppressive activities. In this study, we investigated the effect of rifampicin on atopic dermatitis in vivo and in vitro. AD was induced by treatment with 2, 4-dinitrochlorobenzene (DNCB) in NC/Nga mice. A subset of mice was then treated with rifampicin by oral administration. The severity score and scratching behavior were alleviated in the rifampicin-treated group. Serum immunoglobulin E (IgE) and interleukin-4 (IL-4) levels were also ameliorated in mice treated with rifampicin. We next examined whether rifampicin has anti-atopic activity via suppression of mast cell activation. Rifampicin suppressed the release of β-hexosaminidase and histamine from human mast cell (HMC)-1 cultures stimulated with compound 48/80. Treatment with rifampicin also inhibited secretion of inflammatory mediators, such tumor necrosis factor-α (TNF-α) and prostaglandin D₂ (PGD₂), in mast cells activated by compound 48/80. The mRNA expression of cyclooxygenase 2 (COX-2) was reduced in the cells treated with rifampicin in a concentration-dependent manner. These results suggest that rifampicin can be used to treat atopic dermatitis.

Rifampicin Alleviates Atopic Dermatitis-Like Response in vivo and in vitro

Atopic dermatitis (AD) is a common inflammatory skin disorder mediated by inflammatory cells, such as macrophages and mast cells. Rifampicin is mainly used for the treatment of tuberculosis. Recently, it was reported that rifampicin has anti-inflammatory and immune-suppressive activities. In this study, we investigated the effect of rifampicin on atopic dermatitis in vivo and in vitro. AD was induced by treatment with 2, 4-dinitrochlorobenzene (DNCB) in NC/Nga mice. A subset of mice was then treated with rifampicin by oral administration. The severity score and scratching behavior were alleviated in the rifampicin-treated group. Serum immunoglobulin E (IgE) and interleukin-4 (IL-4) levels were also ameliorated in mice treated with rifampicin. We next examined whether rifampicin has anti-atopic activity via suppression of mast cell activation. Rifampicin suppressed the release of β-hexosaminidase and histamine from human mast cell (HMC)-1 cultures stimulated with compound 48/80. Treatment with rifampicin also inhibited secretion of inflammatory mediators, such tumor necrosis factor-α (TNF-α) and prostaglandin D₂ (PGD₂), in mast cells activated by compound 48/80. The mRNA expression of cyclooxygenase 2 (COX-2) was reduced in the cells treated with rifampicin in a concentration-dependent manner. These results suggest that rifampicin can be used to treat atopic dermatitis.

Curative effect of BCG-polysaccharide nuceic acid on atopic dermatitis in mice

Objective: To explore the effect of bacilli Galmette-Gurin (BCG)-polysaccharide nuceic acid on atopic dermatitis in mice and its mechanism. Method: Forty NC/Nga mice were selected and randomly divided into Group A (model group), Group B (dexamethasone treatment group), Group C (BCG polysaccharide nucleic acid treatment group) and Group D (control group) with 10 mice in each group. Atopic dermatitis model were constructed by applying 2, 4-dinitrochlorobenzene on the skin of the mice. Mice in Group D were treated with acetone solution (100 μL) on the foot pad and abdomen after hair removal at the age of 7 weeks, then on ear skin at the age of 8-13 weeks. For mice in A, B and C groups, 100 μL of acetone solution containing 2, 4-dinitrochlorobenzene was applied to the foot pad and the abdomen at the age of 7 weeks, then on ear skins at the age of 8 to 13 weeks. At the age of 7-13 weeks, mice in Group A and Group D were treated with 100 μL saline (. i.p.); mice were given dexamethasone (0.1 mL/kg, i.p.) every other day for 7 weeks in Group B; mice were treated with BCG polysaccharide nucleic acid (0.5 mg/kg, i.p.) every other day for 7 weeks in Group C. The ear thickness was measured every week and the scratching frequency was recorded 1 times for 10 min a week. The mice were sacrificed after the last administration of drugs. IgE, IL-4, IL-10, IL-12 and IFN-γ in the plasma were detected using ELISA, and RT-PCR method was employed to detect the concentrations of IL-4, IL-10, IL-12 and IFN-γ proteins. After HE staining, the lesion degree of inflammation in ear tissue was observed microscopically. Results: The ear thickness and scratching frequency of Group A were significantly higher than those in group B, C and D (. P0.05); the concentrations of IgE, IL-4 and IL-10 in the plasma and the expression of IL-4, IL-10 mRNA in the spleen tissues of Group A, B and C were all significantly higher than those of Group D (. P<0.05); the concentrations of plasma IL-12 and IFN-γ, and spleen protein expression of IL-12 and IFN-γ in Group C mice were significantly higher than those of Group A (. P<0.05). Histological observation showed obvious ear tissue exudation, erythema, swelling, desquamation of skin, and scabbing in Group A. Histopathology of the skin lesion also showed hyperkeratosis, focal-parakeratosis, stratum spinosum hypertrophy, mild sponge-like edema, a large number of lymphocytes along with plasma cell infiltration in dermis, angiectasis and hyperemia in Group A, while degree of ear skin lesion in Group B and D mice was significantly lighter than that of Group A. Conclusions: BCG polysaccharide nucleic acid can significantly reduce the serum IgE concentrations, increase the expression of IL-12, IFN-γ protein, correct the imbalance of Thl/Th2 in atopic dermatitis mice, and has obvious inhibitory effect on atopic dermatitis in NC/Nga mice.

Curative effect of BCG-polysaccharide nuceic acid on atopic dermatitis in mice

OBJECTIVE@#To explore the effect of bacilli Galmette-Gurin (BCG)-polysaccharide nuceic acid on atopic dermatitis in mice and its mechanism.@*METHOD@#Forty NC/Nga mice were selected and randomly divided into Group A (model group), Group B (dexamethasone treatment group), Group C (BCG polysaccharide nucleic acid treatment group) and Group D (control group) with 10 mice in each group. Atopic dermatitis model were constructed by applying 2, 4-dinitrochlorobenzene on the skin of the mice. Mice in Group D were treated with acetone solution (100 μL) on the foot pad and abdomen after hair removal at the age of 7 weeks, then on ear skin at the age of 8-13 weeks. For mice in A, B and C groups, 100 μL of acetone solution containing 2, 4-dinitrochlorobenzene was applied to the foot pad and the abdomen at the age of 7 weeks, then on ear skins at the age of 8 to 13 weeks. At the age of 7-13 weeks, mice in Group A and Group D were treated with 100 μL saline (i.p.); mice were given dexamethasone (0.1 mL/kg, i.p.) every other day for 7 weeks in Group B; mice were treated with BCG polysaccharide nucleic acid (0.5 mg/kg, i.p.) every other day for 7 weeks in Group C. The ear thickness was measured every week and the scratching frequency was recorded 1 times for 10 min a week. The mice were sacrificed after the last administration of drugs. IgE, IL-4, IL-10, IL-12 and IFN-γ in the plasma were detected using ELISA, and RT-PCR method was employed to detect the concentrations of IL-4, IL-10, IL-12 and IFN-γ proteins. After HE staining, the lesion degree of inflammation in ear tissue was observed microscopically.@*RESULTS@#The ear thickness and scratching frequency of Group A were significantly higher than those in group B, C and D (P0.05); the concentrations of IgE, IL-4 and IL-10 in the plasma and the expression of IL-4, IL-10 mRNA in the spleen tissues of Group A, B and C were all significantly higher than those of Group D (P<0.05); the concentrations of plasma IL-12 and IFN-γ, and spleen protein expression of IL-12 and IFN-γ in Group C mice were significantly higher than those of Group A (P<0.05). Histological observation showed obvious ear tissue exudation, erythema, swelling, desquamation of skin, and scabbing in Group A. Histopathology of the skin lesion also showed hyperkeratosis, focal-parakeratosis, stratum spinosum hypertrophy, mild sponge-like edema, a large number of lymphocytes along with plasma cell infiltration in dermis, angiectasis and hyperemia in Group A, while degree of ear skin lesion in Group B and D mice was significantly lighter than that of Group A.@*CONCLUSIONS@#BCG polysaccharide nucleic acid can significantly reduce the serum IgE concentrations, increase the expression of IL-12, IFN-γ protein, correct the imbalance of Thl/Th2 in atopic dermatitis mice, and has obvious inhibitory effect on atopic dermatitis in NC/Nga mice.

Therapeutic Effects and Immunomodulation of Suanbo Mineral Water Therapy in a Murine Model of Atopic Dermatitis

BACKGROUND: Balneotherapy is widely used as an alternative treatment modality for AD. Although the clinical benefit of some mineral waters has been established, their mechanisms of action in alleviating AD are only partly understood. OBJECTIVE: The clinical modification and immunomodulatory or anti-inflammatory effects of mineral water from the Suanbo hot springs on the differentiation and cytokine production of Th1, Th2, and regulatory T cells (Treg) were investigated using spleen, skin tissue, and serum from NC/Nga mice. METHODS: The therapeutic effects of bathing in mineral water in a Dermatophagoides farinae body extract ointment (Dfb ointment)-induced AD mouse model were assessed by measuring the modified Scoring atopic dermatitis (SCORAD) index scores, transepidermal water loss (TEWL), histological and immunohistochemical changes of the skin lesion, serum levels of interferon (IFN)-gamma, interleukin (IL)-4, IL-5 and immunoglobulin E, mRNA expression of IFN-gamma, IL-4 and IL-5 of dorsal skin, and helper T cell differentiation in the spleen. RESULTS: Bathing in mineral water significantly reduced the modified SCORAD index scores, TEWL, epidermal hyperplasia, and inflammatory cell infiltration. IL-4 production and Th2 cell differentiation showed a decreasing tendency with mineral water bathing, but the Th1 cells did not. On the contrary, differentiation to Treg cells was promoted with mineral water bathing. CONCLUSION: Balneotherapy not only has anti-inflammatory activity, but also shows positive effects on cutaneous barrier homeostasis. These results suggest that the favorable effects of balneotherapy may be mediated by modifying the Th2 response, and possibly in part by inducing Treg cell differentiation.

Dietary effect of silk protein on epidermal levels of free sphingoid bases and phosphate metabolites in NC/Nga mice

In our previous studies, dietary supplements of silk protein, sericin, and fibroin, were beneficial for improving epidermal levels of ceramides, which are the major lipids for maintaining the epidermal barrier. In this study, we investigated the dietary effects of silk protein on epidermal levels of free sphingoid bases and their phosphates such as C18 sphingosine (So), C18 sphinganine (Sa), C18 sphingosine-1-phosphate (S1P), and C18 sphinganine-1-phosphate (Sa1P), which are either synthetic substrate or degradative metabolites of ceramides. Forty-five male NC/Nga mice, an animal model of atopic dermatitis (AD), were divided into three groups: group CA was an atopic control and fed a control diet, group S was fed a 1% sericin diet, and group F was fed a 1% fibroin diet. Fifteen male BALB/c mice served as group C (control group) and were fed the control diet. All mice were fed with diets and water ad libitum for 10 weeks. Sa in group CA was lower than that in group C, but So in group CA was similar to that in group C. So and Sa were higher in groups S and F than those in group CA; So level was even higher than that in group C, and Sa level was similar to that of group C. The So/Sa ratio in group CA, which is reported to increase in AD, was significantly higher than that of group C. The So/Sa ratio was lower in groups S and F than that in group CA, and decreased further in group F. However, S1P and Sa1P in groups S and F were similar to those in group CA. Taken together, we demonstrated that silk protein, sericin and fibroin dietary supplements, increased So and Sa levels, and decreased the So/Sa ratio.

Inhibitory Effects of Korean Red Ginseng Extract on Atopic Dermatitis in NC/Nga Mice / 한국실험동물학회지

Atopic dermatitis (AD) is a chronic eczematous skin disease attended by pruritus, erythema, edema, excoriation, and dryness. This study was to evaluate the effects of Korean red ginseng (RG) on AD in NC/Nga mice treated with 1-chloro-2,4,6-trinitrobenzene (picryl chloride; PC). Experimental groups were divided into 4 groups; normal control (NC), PC control, and PC-RG (50 and 100 mg/kg). RG was orally administered every day repeatedly during 6 weeks. The skin lesions in severity score, scratching behavior, serum immunoglobulin E (IgE), interleukin-4 (IL-4) and interferon-gamma (IFN-gamma) levels, and histological appearance were examined. AD-like lesions were developed on the NC/Nga mice by topical PC applications. Oral administration of RG (50 and 100 mg/kg) significantly suppressed the development of AD, as analyzed by a modified SCORAD score. The scratching behavior decreased after RG administration. The levels of serum IgE, IL-4 and IFN-gamma were increased by PC stimulation, but treatment with RG (100 mg/kg) suppressed the increment of the serum IgE, IL-4 and IFN-gamma levels. Histologically, RG inhibited dermatitis lesions such as hypertrophy, hyperkeratosis, and infiltration of inflammatory cells into epidermis and dermis. These results suggest that the administration of RG may be effective in alleviating the AD induced by PC.

Inhibitory Effects of Korean Red Ginseng Extract on Atopic Dermatitis in NC/Nga Mice / 한국실험동물학회지

Atopic dermatitis (AD) is a chronic eczematous skin disease attended by pruritus, erythema, edema, excoriation, and dryness. This study was to evaluate the effects of Korean red ginseng (RG) on AD in NC/Nga mice treated with 1-chloro-2,4,6-trinitrobenzene (picryl chloride; PC). Experimental groups were divided into 4 groups; normal control (NC), PC control, and PC-RG (50 and 100 mg/kg). RG was orally administered every day repeatedly during 6 weeks. The skin lesions in severity score, scratching behavior, serum immunoglobulin E (IgE), interleukin-4 (IL-4) and interferon-gamma (IFN-gamma) levels, and histological appearance were examined. AD-like lesions were developed on the NC/Nga mice by topical PC applications. Oral administration of RG (50 and 100 mg/kg) significantly suppressed the development of AD, as analyzed by a modified SCORAD score. The scratching behavior decreased after RG administration. The levels of serum IgE, IL-4 and IFN-gamma were increased by PC stimulation, but treatment with RG (100 mg/kg) suppressed the increment of the serum IgE, IL-4 and IFN-gamma levels. Histologically, RG inhibited dermatitis lesions such as hypertrophy, hyperkeratosis, and infiltration of inflammatory cells into epidermis and dermis. These results suggest that the administration of RG may be effective in alleviating the AD induced by PC.

Inhibitory effects of interleukin-10 plasmid DNA on the development of atopic dermatitis-like skin lesions in NC/Nga mice

Interleukin (IL)-10 exerts potent anti-inflammatory effects by suppression of both T-help (Th) 1 and Th2 cells. Previous studies have reported that IL-10 can ameliorate various inflammatory disorders. The present study was performed to examine whether IL-10 plasmid DNA could suppress development of atopic dermatitis (AD)-like skin lesions in NC/Nga mice, as an initial step towards the development of an appliance for use in dogs with AD. Intradermal injection of IL-10 plasmid DNA markedly inhibited the development of AD-like skin lesions, as evidenced by a marked decrease in skin symptoms and reduced inflammation within the skin lesions. Efficacy was confirmed by significant decreases in eosinophil ratio and serum IgE concentration, and a reduction in the number of Staphylococcus aureus recovered from the ear. Moreover, relative mRNA expression levels of IL-4 and interferon-gamma in the skin lesions of mice injected with IL-10 plasmid DNA were also decreased compared with those of control mice. Of note, higher serum IL-10 levels in mice injected with IL-10 plasmid DNA were maintained compared with those in control mice. Taken together, the results indicate that IL-10 plasmid DNA can suppress the development of AD-like skin lesions by suppressing both Th1 and Th2 cell responses. Beneficial effects of IL-10 plasmid DNA may be expected in dogs with AD.

Developing an Atopic Dermatitis Model and the Effects of Actinidia Extract on Dermatitis in NC/Nga Mice / 대한피부과학회지

BACKGROUND: Atopic dermatitis is a chronic itchy, inflammatory skin disease that usually relapses. Although the etiology of atopic dermatitis remains unclear, it has been shown that both Th1 and Th2 cytokines play pathogenic roles in the generation of atopic dermatitis. DA-9102 is a fraction from the Actinidia species and DA-9102 displays immune modulating activity for allergy related disease. OBJECTIVE: We have developed the atopic dermatitis model of NC/Nga mice using DNCB and we examined whether DA-9102 suppresses the development of atopic dermatitis-like skin lesions on NC/Nga mice. METHODS: NC/Nga mice were challenged with DNCB during 5 weeks to develop atopic dermatitis-like skin lesions. Daily DA-9102 or cyclosporine A or HPMC (control) were then given orally. The efficacy of DA-9102 in NC/Nga mice was judged by measurement of the skin lesion severity (a modified SCORAD score), the serum IgE and IgG2a levels and the cytokine levels (IFN-gamma and IL-4) from spleen cells cultured with ConA. RESULTS: Atopic dermatitis-like lesions were developed on the NC/Nga mice by using topical DNCB. Oral administration of 100 mg/kg DA-9102 significantly suppressed the development of dermatitis, as was analyzed by a modified SCORAD score (p<0.01). The serum IgE level increased gradually with age, but treatment with DA-9102 suppressed the increment of the serum IgE level (p<0.01). The mean values of IFN-gamma in the NC/Nga mice of the DA-9102 group were lower than those of the control mice group (p<0.05). The mean values of IL-4 were undetectable in all the experimental groups. The serum IgG2a level were not significantly different among all the experimental groups. CONCLUSION: We successfully developed an atopic dermatitis model in NC/Nga mice. Based on our in in vitro data, we suggest that DA-9102 can be useful for the treatment of atopic dermatitis.

Dietary Effect of Silk Protein Sericin or Fibroin on Plasma and Epidermal Amino Acid Concentration of NC/Nga Mice

Free amino acids in epidermis function as a major component of Natural Moisturizing Factor (NMF), which maintains the optimal level of water in skin even at the low humidity. In fact, the depletion of free amino acids is reported in the epidermis of atopic dermatitis, the skin condition involving dryness. As an effort searching the dietary source for improving the level of water and free amino acid in epidermis, the dietary effects of silk protein, sericin (S) and fibroin (F) on trans epidermal water loss (TEWL), and plasma and epidermal levels of free amino acids were compared in this study. Thirty of male NC/Nga mice, an animal model of atopic dermatitis, were divided into three groups: group CA as an atopic control with control diet, group S: 1% sericin diet and group F: 1% fibroin diet. Ten of male BALB/c mice were served as group C (control group) with control diet. All mice were fed on diet and water ad libitum for 10weeks. Dry skin condition was established in group CA as TEWL was increased (148.7% of group C). In parallel, epidermal level of glutamate, one of major amino acids functioning as NMF, was dramatically decreased and epidermal levels of methionine and alanine were inversely elevated. Dietary supplementation of sericin (group S) reduced TEWL at the similar level with group C and increased epidermal levels of glutamate as well as serine and glycine, the other major amino acids as NMF. Despite a marked decrease of methionine and alanine, the reduction of TEWL and epidermal levels of glutamate, serine and glycine of group F were less than of group S. Furthermore, in contrast to similar levels of other free amino acids in plasma and epidermis of group S and group C, plasma and epidermal levels of other free amino acids, specifically phenylalanine, isoleucine, cysteine and tyrosine in epidermis of group F, were significantly higher than of group C. Together, our data demonstrate that dietary supplementation of sericin is more effective at improving dry skin condition that paralleled with the normalization of free amino acids in plasma and epidermis of NC/Nga mice.

Alteration of Innate Immune T and B Cells in the NC/Nga Mouse

BACKGROUND: Millions of people in the world are suffering from atopic dermatitis (AD), which is a chronic inflammatory skin disease triggered by Th2 immune responses. The NC/Nga mouse is the most extensively studied animal model of AD. Like human AD, NC/Nga mice demonstrate increased levels of IgE, a hallmark of Th2 immune responses. Adaptive immunity cannot be generated without help of innate immunity. Especially natural killer T (NKT) cells and marginal zone B (MZB) cells have been known to play important roles in linking innate immunity to adaptive immunity. METHODS: Through flow cytometric analysis and ELISA assay, we investigated whether these lymphocytes might be altered in number in NC/Nga mice. RESULTS: Our data demonstrated that the number of NKT cells was reduced in NC/Nga mice and IFNgamma production by NKT cells upon alpha-GalCer stimulation decreased to the levels of CD1d KO mice lacking in NKT cells. However, reduction of NKT cells in NC/Nga mice was not due to CD1d expression, which was normal in the thymus. Interestingly, there was a significant increase of CD1d(high)B220+ cells in the spleen of NC/Nga mice. Further, we confirmed that CD1d(high)B220+ cells are B cells, not dendritic cells. These CD1d(high)B220+ B cells show IgM(high)CD21(high)CD23low, a characteristic phenotype of MZB cells. CONCLUSION: We provide the evidence that there are decreased activities of NKT cells and increased number of MZB cells in the NC/Nga mice. Our findings may thus explain why NC/Nga mice are susceptible to AD.