中华皮肤科杂志 ›› 2019, Vol. 52 ›› Issue (7): 481-485.doi: 10.3760/cma.j.issn.0412-4030.2019.07.007

• 论著 • 上一篇    下一篇

卵清蛋白联合卡泊三醇诱导构建特应性皮炎小鼠模型

张宇    韩悦    徐蓓蕾    凌诗琪    罗阳    刘笑纯    姚煦   

  1. 中国医学科学院  北京协和医学院  皮肤病医院过敏与风湿免疫科,南京  210042
  • 收稿日期:2018-09-30 修回日期:2019-05-07 出版日期:2019-07-15 发布日期:2019-07-01
  • 通讯作者: 姚煦 E-mail:dryao_xu@126.com
  • 基金资助:
    国家自然科学基金(81703126);江苏省自然科学基金 (BK20170162)

Establishment of a mouse model of atopic dermatitis induced by topical application of ovalbumin and calcipotriol

Zhang Yu, Han Yue, Xu Beilei, Ling Shiqi, Luo Yang, Liu Xiaochun, Yao Xu   

  1. Department of Allergy and Rheumatology, Hospital for Skin Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Nanjing 210042, China
  • Received:2018-09-30 Revised:2019-05-07 Online:2019-07-15 Published:2019-07-01
  • Contact: Yao Xu E-mail:dryao_xu@126.com
  • Supported by:
    National Natural Science Foundation of China(81703126);Natural Science Foundation of Jiangsu Province of China (BK20170162)

摘要: 【摘要】 目的 探讨快速建立特应性皮炎(AD)小鼠模型的方法。方法 以C57BL/6小鼠为模式动物,利用简单随机化方法分为3组,卡泊三醇 + 卵清蛋白(OVA)组(6只)两侧耳部外涂卡泊三醇和OVA,卡泊三醇组(6只)耳部外涂卡泊三醇,对照组(3只)耳部外涂75%乙醇,连续12 d。分别在造模前和第14天时拍摄小鼠耳部照片并测量小鼠耳厚度,取小鼠尾静脉血,检测血清总IgE和OVA特异性IgE水平,并在第14天取小鼠耳廓皮肤进行组织病理检查。结果 第14天时卡泊三醇 + OVA组和 卡泊三醇组小鼠耳部皮肤出现红肿、干燥、脱屑,耳厚度较造模前均明显增加(均P < 0.001),但两组间耳厚度[分别为(0.355 ± 0.03)和(0.370 ± 0.05) mm]差异无统计学意义(q = 0.674,P = 0.231)。耳部皮肤病理检查显示,与卡泊三醇组和对照组相比,卡泊三醇 + OVA组小鼠表皮增生和炎症细胞(包括嗜酸性粒细胞和肥大细胞)浸润更加明显。皮肤免疫组化检查显示,3组间表皮炎症因子胸腺基质淋巴细胞生成素(TSLP)和干扰素γ表达差异无统计学意义(均P > 0.05),但IL-13差异有统计学意义(F = 5.159,P = 0.032),卡泊三醇 + OVA组IL-13表达水平(77.12 ± 5.46)高于对照组(55.49 ± 9.92,q = 3.170,P = 0.021)。第14天时卡泊三醇 + OVA组和卡泊三醇组小鼠血清总IgE与造模前比较均明显升高,其中卡泊三醇 + OVA组总IgE升高更明显[(8 278.56 ± 3 297.68)比(892.64 ± 82.83) μg/L,t = 4.132,P = 0.026],且卡泊三醇 + OVA组血清OVA特异性IgE水平(192.846 ± 15.391) μg/L显著高于卡泊三醇组[(8.492 ± 3.879) μg/L,q = 22.476,P < 0.001]。 结论 连续12 d外涂卡泊三醇和OVA可快速建立过敏原诱发的AD小鼠模型,为进行过敏原相关的AD发病机制研究奠定了一定基础。

关键词: 皮炎, 特应性; 疾病模型, 动物; 卵白蛋白; 卡泊三醇; 免疫球蛋白E

Abstract: 【Abstract】 Objective To explore a method for rapidly establishing a mouse model of atopic dermatitis (AD). Methods C57BL/6 mice served as model animals, and were randomly divided into 3 groups: calcipotriol + ovalbumin (OVA) group (n = 6) topically treated with calcipotriol and OVA on the mouse ears, calcipotriol group (n = 6) topically treated with calcipotriol on the ears, and control group (n = 3) topically treated with 75% alcohol on the ears. The treatment lasted 12 days. Before the model establishment and on day 14, the photos of the mouse ears were taken, and ear thickness was measured; moreover, blood samples were obtained from the mouse caudal vein, and serum levels of total IgE and OVA-specific IgE were detected. On day 14, the skin tissues of mouse auricles were resected and subjected to histopathological examination. Results On day 14, erythematous swelling, dryness and desquamation occurred on the mouse ear skin in the calcipotriol + OVA group and calcipotriol group, and both the two groups showed significantly increased ear thickness compared with those before the model establishment (both P < 0.001). However, there was no significant difference in the ear thickness between the calcipotriol + OVA group (0.355 ± 0.03 mm) and calcipotriol group (0.370 ± 0.05 mm, q = 0.674, P = 0.231). Histopathological examination of the ear skin showed more obvious epidermal hyperplasia and infiltration of dermal inflammatory cells including eosinophils and mastocytes in the calcipotriol + OVA group compared with the calcipotriol group and control group. Immunohistochemical study revealed that there was no significant difference in the expression of thymic stromal lymphopoietin (TSLP) and interferon (IFN)-γ among the 3 groups (both P > 0.05), while the expression of interleukin (IL)-13 significantly differed among the 3 groups (F = 5.159, P = 0.032), and was significantly higher in the calcipotriol + OVA group (77.12 ± 5.46) than in the control group (55.49 ± 9.92, q = 3.170, P = 0.021). On day 14, the calcipotriol + OVA group and calcipotriol group both showed markedly increased total serum IgE levels compared with those before the treatment, and the calcipotriol + OVA group showed a more significant increase (8 278.56 ± 3 297.68 vs. 892.64 ± 82.83 μg/L, t = 4.132, P = 0.026). Meanwhile, the serum level of OVA-specific IgE was significantly higher in the calcipotriol + OVA group (192.846 ± 15.391 μg/L) than in the calcipotriol group (8.492 ± 3.879 μg/L, q = 22.476, P < 0.001) on day 14. Conclusion The mouse model of allergen-induced AD can be rapidly established by topical application of calcipotriol and OVA for 12 consecutive days, which lays a foundation for further study on allergen-related pathogenesis of AD.

Key words: Dermatitis, atopic, Disease models, animal, Ovalbumin, Calcipotriol, Immunoglobulin E