Skin Lesions In Mouse Model Research Articles

Protocol to Induce Follicular T Helper Cells, Germinal Centers, and Skin Lesions in Mouse Models for Skin Blistering Diseases.

Autoreactive T cells in autoantibody-mediated autoimmune diseases can be divided into two major subsets: (i) follicular T helper cells (Tfh) that provide T cell help in germinal centers (GC) and (ii) effector T (Teff) cells that immigrate into peripheral tissue sites such as the skin and mediate local inflammation. To study the sequence of events leading to the loss of tolerance in autoantibody-mediated autoimmune diseases it is required to investigate both T cell subsets simultaneously. This approach is hampered mainly because the appearance of skin inflammation in mouse models is a random process, which makes it difficult to define the location of inflammation at the right time point. To overcome this problem, we developed a scratching technique for ear skins that leads to the establishment of chronic autoimmune wounds in the mouse model for the pemphigoid-like disease epidermolysis bullosa acquisita. By defining the exact place where the skin wounds should form, this protocol enables a detailed analysis of skin-immigrating Teff cells. Of note, this protocol induces GC in draining lymph nodes in parallel so that Tfh cells in GC can be investigated concurrently. This protocol is not restricted to T cells and can be used for any other skin-immigrating inflammatory cells.

Therapeutic effects of shikonin on skin lesions in mouse models of allergic dermatitis and wound

ABSTRACTAimThe aim of this study was to examine the beneficial effects of shikonin on skin lesions in mouse models of allergic dermatitis and wound.MethodsMouse models of allergic dermatitis and wound were generated by spreading an allergic‐inducing reagent on the dorsal skin and earflaps, and by punching the dorsal skin, respectively. The effects of shikonin on allergic dermatitis was evaluated on frequency of scratching, clinical severity of dermatitis and immunohistochemistry. The effect of shikonin on wound healing compared with steroid was evaluated by the size of the wound area.ResultsShikonin treatment not only improved allergic dermatitis, but also prevented the onset of allergic dermatitis. Furthermore, shikonin promoted wound healing in a mouse model of skin wound. In contrast, steroid (betamethasone) treatment, which is often applied to treat allergic dermatitis, delayed wound healing in the mouse model of allergic dermatitis.ConclusionShikonin can be used as a new therapeutic agent for the treatment of allergic dermatitis including atopic dermatitis.

Effects of extracts of Semen Coicis on a BALB/c mouse model of atopic dermatitis and its mechanism

Objective To evaluate the effects of extracts of Semen Coicis (ESC) on a BALB/c mouse model of atopic dermatitis (AD) , and to explore its potential mechanism. Methods Forty specific pathogen-free (SPF) female BABL/c mice were randomly divided into blank group (8 mice, receiving no treatment) and AD model group (32 mice) . The mice in the model group were topically treated with 2,4-dinitrochlorobenzene (DNCB) in acetone/olive oil to establish the mouse model of AD. After modeling, 8 mice in the blank group and 8 in the model group were sacrificed immediately. The other 24 mice in the model group were randomly and equally divided into 3 groups: model control group receiving no treatment, ESC group and ESC vehicle group topically treated with ESC and ESC vehicle respectively once every day on the back and aural region of the mice for 28 consecutive days. Changes in skin lesions were observed by naked eyes every day. A thickness tester was used to measure the thickness of skin lesions on the left ear before modeling, at completion of modeling and 12 hours after the final treatment. At 12 hours after the final treatment, the mice in the above 3 groups were sacrificed, and the eyeballs were removed for collecting blood. Then, the sera were isolated, and skin tissue specimens were obtained from the skin lesions on the back. These tissue sections were subjected to hematoxylin and eosin (HE) staining and toluidine blue staining for observing the infiltration of inflammatory cells in skin lesions. An immunohistochemical study was performed to determine the expression of aquaporin 3 (AQP3) , Toll-like receptor 2 (TLR2) and TLR4, and enzyme-linked immunosorbent assay (ELISA) to detect the serum levels of IgE, interleukin-4 (IL-4) and interferon-γ (IFN-γ) . Results After 28-day treatment, skin lesions were improved in the ESC group. Compared with the model control group, the ESC group showed a significantly lower clinical symptom score (1.50 ± 0.58 vs. 2.50 ± 0.58, P < 0.05) , decreased lesional thickness on the left ear ([0.31 ± 0.01] mm vs. [0.33 ± 0.01] mm, P < 0.05) , and lower number of infiltrating mast cells per high-power field (15.18 ± 1.64 vs. 28.94 ± 1.28, P < 0.05) . Immunohistochemical findings indicated that the ESC group showed significantly lower expression of AQP3, TLR2 and TLR4 compared with the model control group, and decreased AQP3 expression in the spinous layer. Compared with the model control group, the ESC group showed significantly lower total serum IgE and IL-4 levels, but higher IFN-γ levels (all P < 0.05) . Conclusion Topical ESC is effective for the treatment of skin lesions in mouse models of AD, likely by regulating serum levels of IgE, IL-4 and IFN-γ and affecting the expression of AQP3, TLR2 and TLR4. Key words: Dermatitis, atopic; Semen Coicis; Disease models, animal; Interleukin-4; Interferon-gamma; Toll-like receptors; Aquaporin 3

Effects of Twelve Korean Combined Herbal Prescriptions with Platycodon Grandiflorum on Induction of Autophagy and Inhibition of Cell Proliferation

Among the 22 included studies, 21 herbs were reported to reduce AD-like skin lesions in mouse models by suppressing Th2 cell responses. Conclusion:By summarizing the results fromthepublished literature,wehope that this studymight aid in finding a potential herbal therapeutic agent for the treatment of AD. The limitation of this study was that a meta-analysis was not conducted because of the variety of investigated herbs included in the studies. Nevertheless, this reviewmay assist in identifying directions for further researches endeavors. Contact: Younghee Yun, allergycosmetic@khnmc.or.kr

Topical herbal application in the management of atopic dermatitis: a review of animal studies.

Herbs are widely used in the treatment of atopic dermatitis (AD) in Eastern Asian countries, and certain herbs regarded have anti-inflammatory properties that can help with AD. With the goal of developing a topical herbal agent for AD, we conducted a systematic review of in vivo studies of AD-like skin models for screening potential herbs. Searches were conducted from PubMed and EMBASE. After all, 22 studies were included for this review. We judged most of the domains of all studies to be at unclear risk of bias. Among 22 included studies, 21 herbs have been reported to reduce AD-like skin lesions in mouse models by suppressing Th2 cell response. Our findings may offer potential herbs for the topical application treatment of AD.

An attempt to develop mouse model for anti-laminin γ1 pemphigoid

BackgroundWe recently reported that the autoantibodies of anti-p200 pemphigoid sera react with laminin γ1 and renamed this entity as anti-laminin γ1 pemphigoid. However, it has not been clarified whether the anti-laminin γ1 autoantibodies, particularly those to the C-terminal integrin binding site, affect the dermoepidermal junction and cause subepidermal blisters. ObjectiveThe aim of this study was to develop animal models for anti-laminin γ1 pemphigoid. MethodsWe attempted to produce two mouse models for anti-laminin γ1 pemphigoid; (1) a passive transfer model: injection of rabbit IgG to shorter bacterial recombinant protein of the murine laminin γ1 C-terminal 107 amino acids, and (2) an active disease model: direct immunization to mice with this recombinant protein. ResultsImmunoblotting revealed that 70% of patient sera reacted with the shorter recombinant protein of human laminin γ1 C-terminus. In the passive transfer model, rabbit IgG to the murine laminin γ1 C-terminus was deposited, without C3 deposition, at the epidermal basement membrane zone. In contrast, in the active disease model, direct immunofluorescence of mouse skin sections showed no deposition of either murine IgG or C3. Blister formation was not seen in either model both phenotypically and histopathologically. ConclusionIn the two different mouse animal models for anti-laminin γ1 pemphigoid, although rabbit IgG to the recombinant laminin γ1 C-terminus bound to the epidermal basement membrane zone in passive transfer model, no obvious blister formation was seen. To reproduce skin lesions in mouse models for anti-laminin γ1 pemphigoid, further improvement should be needed.

Time course pathogenesis of sulphur mustard‐induced skin lesions in mouse model

Sulphur mustard (SM) is a bifunctional alkylating agent that causes cutaneous blistering in humans and animals. In this study, we have presented closer views on pathogenesis of SM-induced skin injury in a mouse model. SM diluted in acetone was applied once dermally at a dose of 5 or 10 mg/kg to Swiss albino mice. Skin was dissected out at 0, 1, 3, 6, 12, 24, 48, 72 and 168 hours, post-SM exposure for studying histopathological changes and immunohistochemistry of inflammatory-reparative biomarkers, namely, transforming growth factor alpha (TGF-α), fibroblast growth factor (FGF), endothelial nitric oxide synthase (eNOS) and interlukin 6 (IL-6). Histopathological changes were similar to other mammalian species and basal cell damage resembled the histopathological signs observed with vesication in human skin. Inflammatory cell recruitment at the site of injury was supported by differential expressions of IL-6 at various stages. Time-dependent expressions of eNOS played pivotal roles in all the events of wound healing of SM-induced skin lesions. TGF-α and FGF were strongly associated with keratinocyte migration, re-epithelialisation, angiogenesis, fibroblast proliferation and cell differentiation. Furthermore, quantification of the tissue leukocytosis and DNA damage along with semiquantitative estimation of re-epithelialisation, fibroplasia and neovascularisation on histomorphologic scale could be efficiently used for screening the efficacy of orphan drugs against SM-induced skin injury.

Inhibition of atopic dermatitis‐like skin lesions by topical application of a novel ceramide derivative, K6PC‐9p, in NC/Nga mice

Atopic dermatitis (AD) is a chronic inflammatory skin disease that commonly begins in childhood. K6PC-9p (N-(Ethyl dihydrogenphosphate)-2-hexyl-3-oxo-decanamide) is a synthetic ceramide derivative of PC-9S (N-Ethanol-2-mirystyl-3-oxo-staramide), which was known to be effective in atopic patients. In this study, we examined the effect of topical application of K6PC-9p on skin inflammation and AD-like skin lesions in mouse models. K6PC-9p dose-dependently inhibited phorbol ester-induced increase in ear thickness in BALB/c mice. Moreover, topical application of K6PC-9p suppressed dust mite extract-induced AD-like skin lesions in NC/Nga mice. Histopathological analysis revealed that both ear swelling and leucocyte infiltration were suppressed by K6PC-9p treatment. K6PC-9p also suppressed IL-4 and TNF-alpha expression in the ears and mast cell infiltration into the ears in NC/Nga mice. Further study demonstrated that K6PC-9p inhibited ConA-induced IL-4 secretion and LPS-induced macrophage activation. Taken together, our results showed that topical application of K6PC-9p exerts beneficial effects in animal model of skin inflammation and AD, suggesting that K6PC-9p might be a promising topical agent for the treatment of inflammatory skin diseases.

Topical application of a novel ceramide derivative, K6PC-9, inhibits dust mite extract-induced atopic dermatitis-like skin lesions in NC/Nga mice

Atopic dermatitis (AD) is a chronic inflammatory skin disease. K6PC-9 (N-Ethanol-2-hexyl-3-oxo-decanamide) is a novel synthetic ceramide derivative of PC-9S (N-Ethanol-2-mirystyl-3-oxo-stearamide), which was known to be effective in atopic and psoriatic patients. To investigate the immunomodulatory activity of K6PC-9, we examined the effect of K6PC-9 on T lymphocyte and macrophage function and the effect of topical application of K6PC-9 on skin inflammation and AD-like skin lesions in mouse models. K6PC-9 had no effect on concanavalin A-induced proliferation, interleukin (IL)-2 secretion and IL-4 secretion in mouse splenocytes. In contrast, lipopolysaccharide-induced nitrite generation was potently suppressed by K6PC-9 in mouse peritoneal macrophages. In mouse model of skin inflammation, K6PC-9 inhibited phorbol ester-induced increase in ear thickness and expression of tumor necrosis factor-α in the ear of BALB/c mice. Topical application of K6PC-9 also suppressed mite extract-induced AD-like skin lesions in NC/Nga mice. Increase in ear thickness was significantly inhibited by K6PC-9 in this model. K6PC-9 also blocked the infiltration of mast cells and neutrophils into the ear. Further study demonstrated that the mRNA expression of tumor necrosis factor-α and adhesion molecules, such as vascular cell adhesion molecule-1, intercellular adhesion molecule-1, E-selectin, was also suppressed by K6PC-9 in the ear of mite extract-treated NC/Nga mice. Taken together, the results presented in this report show that K6PC-9 has an anti-inflammatory potential and exerts beneficial effects in an animal model of AD, indicating that K6PC-9 might be used as a topical agent for the treatment of AD.