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特应性皮炎犬类动物模型皮肤中丝聚合蛋白的mRNA和蛋白质表达改变。

Altered mRNA and protein expression of filaggrin in the skin of a canine animal model for atopic dermatitis.

作者信息

Santoro Domenico, Marsella Rosanna, Ahrens Kim, Graves Thomas K, Bunick David

机构信息

Department of Veterinary Clinical Medicine, University of Illinois at Urbana-Champaign, 1008 W Hazelwood Drive, Urbana, IL 61802, USA.

出版信息

Vet Dermatol. 2013 Jun;24(3):329-36, e73. doi: 10.1111/vde.12031.

Abstract

BACKGROUND

Filaggrin is a structural protein that has attracted increasing interest over the past decade for its role in the pathogenesis of human atopic dermatitis (AD). Null mutations in its sequence are considered risk factors in the development of AD.

HYPOTHESIS/OBJECTIVES: To investigate canine filaggrin mRNA and protein expression in the skin of atopic beagles with experimentally induced AD compared with breed-matched healthy control dogs.

METHODS

All dogs were environmentally challenged for 3 days consecutively with allergens to which the atopic dogs had been sensitized. Skin biopsy specimens were taken from six healthy and seven atopic beagles before and after allergen challenge. Canine filaggrin mRNA was measured using quantitative real-time PCR. Indirect immunofluorescence was used to localize the filaggrin protein in canine skin. Analysis of variance with Tukey's multiple comparison test (over-time effect) and unpaired Student's t-test (treatment effect) were used. Values of P ≤ 0.05 were considered significant.

RESULTS

Analysis of variance showed a significantly higher expression of filaggrin mRNA in atopic dogs compared with healthy control dogs (P = 0.004 on day 3 and P = 0.01 on day 10) and a decreased mRNA expression on day 3 in healthy control dogs (effect of time, P = 0.006). On blinded evaluation, filaggrin immunofluorescence was distributed homogeneously in the stratum granulosum and the stratum corneum in healthy dogs. Atopic dogs showed a patchy immunofluorescence pattern, which was exacerbated after environmental challenge.

CONCLUSIONS AND CLINICAL IMPORTANCE

Altered epidermal filaggrin mRNA expression and protein distribution was detected in this experimental model.

摘要

背景

丝聚合蛋白是一种结构蛋白,在过去十年中,因其在人类特应性皮炎(AD)发病机制中的作用而受到越来越多的关注。其序列中的无效突变被认为是AD发病的危险因素。

假设/目的:研究与品种匹配的健康对照犬相比,实验性诱导AD的特应性比格犬皮肤中犬丝聚合蛋白mRNA和蛋白的表达情况。

方法

所有犬只连续3天接受特应性犬已致敏的过敏原环境刺激。在过敏原刺激前后,从6只健康比格犬和7只特应性比格犬身上采集皮肤活检标本。使用定量实时PCR测量犬丝聚合蛋白mRNA。采用间接免疫荧光法在犬皮肤中定位丝聚合蛋白。使用方差分析和Tukey多重比较检验(时间效应)以及未配对学生t检验(治疗效应)。P≤0.05的值被认为具有统计学意义。

结果

方差分析显示,与健康对照犬相比,特应性犬丝聚合蛋白mRNA的表达显著更高(第3天P = 0.004,第10天P = 0.01),且健康对照犬在第3天mRNA表达下降(时间效应,P = 0.006)。在盲法评估中,丝聚合蛋白免疫荧光在健康犬的颗粒层和角质层中均匀分布。特应性犬呈现出斑片状免疫荧光模式,在环境刺激后加剧。

结论及临床意义

在该实验模型中检测到表皮丝聚合蛋白mRNA表达和蛋白分布改变。

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