中华皮肤科杂志 ›› 2018, Vol. 51 ›› Issue (8): 609-613.doi: 10.3760/cma.j.issn.0412-4030.2018.08.014

• 研究报道 • 上一篇    下一篇

薏苡仁提取物对BALB/c小鼠特应性皮炎模型的疗效观察及机制探讨

王俊霞1,杨子微1,单士军2,车雅敏1,陈洪铎3   

  1. 1. 天津医科大学总医院
    2. 天津
    3. 中国医科大学第一附属医院皮肤科
  • 收稿日期:2018-01-15 修回日期:2018-06-29 出版日期:2018-08-15 发布日期:2018-07-31
  • 通讯作者: 单士军 E-mail:15822183620@163.com

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

Wang Junxia, Yang Ziwei, Che Yamin, Shan Shijun, Chen Hongduo   

  1. Department of Dermatology and Venereology, Tianjin Medical University General Hospital, Tianjin 300052, China (Wang JX, Yang ZW, Che YM, Shan SJ); Department of Dermatology and Venereology, China Medical University, The First Hospital of China Medical University, Shenyang 110001, China (Chen HD)
  • Received:2018-01-15 Revised:2018-06-29 Online:2018-08-15 Published:2018-07-31
  • Contact: Shan Shijun E-mail:15822183620@163.com

摘要: 目的 观察薏苡仁提取物(ESC)对BALB/c特应性皮炎(AD)模型小鼠的疗效,探讨其可能的作用机制。方法 SPF级雌性纯系BALB/c小鼠40只,随机分为空白组(8只)和模型组(32只)。模型组应用2,4-二硝基氯苯(DNCB)和丙酮、橄榄油溶液构建AD模型。造模完成后,空白组8只、模型组8只小鼠立即处死,模型组另24只小鼠随机分为模型对照组、ESC组、基质组3组。模型对照组不予任何处理,ESC组、基质组背部及耳部分别外涂薏苡仁提取物和基质每日1次,连续28 d。每天肉眼观察皮损变化;测厚仪检测造模前、造模完成时及末次给药后12 h小鼠左耳皮损厚度;末次给药后12 h,处死3组小鼠,摘眼球取血,分离血清,并于背部皮损处取组织标本。组织切片后行HE染色和甲苯胺蓝染色观察皮损炎症细胞浸润情况;免疫组化法检测水通道蛋白3(AQP3)、Toll样受体(TLR)2、4表达变化;ELISA法检测血清IgE、白细胞介素4(IL-4)和干扰素γ(IFN-γ)水平。结果 治疗28 d后,ESC组小鼠皮损好转,其临床症状评分(1.50 ± 0.58)低于模型对照组(2.50 ± 0.58)(P < 0.05),左耳部皮损厚度[(0.31 ± 0.01) mm]低于模型对照组[(0.33 ± 0.01) mm](P < 0.05),每高倍视野下浸润的肥大细胞数亦低于模型对照组(28.94 ± 1.28)(P < 0.05)。免疫组化示,ESC组水通道蛋白3(AQP3)及TLR2和TLR4表达水平低于模型对照组,AQP3棘层表达减少。ESC组小鼠血中总IgE、IL-4水平低于模型对照组,IFN-γ水平高于模型对照组,差异均有统计学意义(P < 0.05)。结论 外用薏苡仁提取物对特应性皮炎模型小鼠皮损有治疗作用,可能是通过调节血清IgE、IL-4及IFN-γ水平和影响AQP3 及TLR2和TLR4表达发挥作用。

关键词: 皮炎, 特应性, 薏苡仁, 疾病模型, 动物, 白细胞介素4, 干扰素γ, Toll样受体, 水通道蛋白质3

Abstract: Wang Junxia, Yang Ziwei, Che Yamin, Shan Shijun, Chen Hongduo Department of Dermatology and Venereology, Tianjin Medical University General Hospital, Tianjin 300052, China (Wang JX, Yang ZW, Che YM, Shan SJ); Department of Dermatology and Venereology, China Medical University, The First Hospital of China Medical University, Shenyang 110001, China (Chen HD) Corresponding author: Shan Shijun, Email: 15822183620@163.com 【Abstract】 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 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 of AQP3, TLR2 and TLR4 compared with the model control group, and decreased AQP3 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 of AQP3, TLR2 and TLR4.

Key words: Dermatitis, atopic, Semen Coicis, Disease models, animal, Interleukin-4, Interferon-gamma, Toll-like receptors, Aquaporin 3