中华皮肤科杂志 ›› 2021, Vol. 54 ›› Issue (8): 696-701.doi: 10.35541/cjd.20201084

• 论著 • 上一篇    下一篇

外用紫草素对咪喹莫特诱导银屑病样小鼠模型的干预作用及对CEBPD表达的影响

兰晓鸥1    王强1    李上云1    孙晓冬1    宋丹阳1    李佳蔚2    宋国强1    胡兰廷1    耿龙3   

  1. 1沈阳市第七人民医院皮肤科  110003；2中国医科大学附属盛京医院皮肤科，沈阳  110004；3中国医科大学附属第一医院皮肤科，沈阳  110001
  • 收稿日期:2020-11-12 修回日期:2021-05-27 发布日期:2021-08-02
  • 通讯作者: 耿龙；王强 E-mail:genglong99@126.com; swepco@126.com
  • 基金资助:
    国家自然科学基金（81673055）；辽宁省自然科学基金（2020-MS-325）

Topical shikonin: intervention effect on an imiquimod-induced psoriasis-like mouse model and influence on CEBPD expression

Lan Xiaoou1, Wang Qiang1, Li Shangyun1, Sun Xiaodong1, Song Danyang1, Li Jiawei2, Song Guoqiang1, Hu Lanting1, Geng Long3   

  1. 1Department of Dermatology, The Seventh People′s Hospital of Shenyang, Liaoning 110003, China; 2Department of Dermatology, Shengjng Hospital of China Medical University, Liaoning 110004, China; 3Department of Dermatology, The First Hospital of China Medical University, Liaoning 110001, China
  • Received:2020-11-12 Revised:2021-05-27 Published:2021-08-02
  • Contact: Geng Long; Wang Qiang E-mail:genglong99@126.com; swepco@126.com
  • Supported by:
    National Natural Science Foundation of China （81673055）; Natural Science Foundation of Liaoning Province of China （2020-MS-325）

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摘要： 【摘要】 目的 探讨外用紫草素对咪喹莫特诱导的银屑病样小鼠模型的干预作用及对CCAAT增强子结合蛋白δ（CEBPD）表达的影响。方法 将20只SPF级BALB/c雄性小鼠按照简单随机抽样法分为模型组、紫草素1组、紫草素2组及空白对照组，每组5只。模型组、紫草素1组和紫草素2组小鼠背部脱毛区均每日外涂5%咪喹莫特乳膏（IMQ）50 mg建立银屑病样小鼠模型，用药6 h后，紫草素1组、紫草素2组小鼠建模区分别予0.5 ml浓度为0.576 g/L和5.76 g/L的紫草素，共处理8 d；空白对照组不予任何处理。每日肉眼观察各组小鼠银屑病皮损严重程度指数（PASI）随时间的变化情况；8 d后，脱颈处死小鼠，取背部皮损，通过免疫组化及Western印迹法检测CEBPD在小鼠背部表皮层的表达。采用SPSS 16.0进行统计学分析，不同组间观察指标的比较采用单因素方差分析，两两多重比较采用LSD-t检验。结果 第8天时，模型组小鼠可见明显的红斑、鳞屑及皮损浸润增厚，紫草素1、紫草素2组小鼠与同期模型组相比，皮损显著改善，紫草素2组改善更明显。第8天时，空白对照组PASI评分为0分，模型组为（11.0 ± 1.22）分，紫草素1组为（8.6 ± 0.55）分，紫草素2组为（5.8 ± 1.30）分，各组间差异有统计学意义（F = 128.21，P＜0.01），组间两两比较显示差异亦均有统计学意义（均P＜0.01）。免疫组化显示，模型组CEBPD表达水平（A值）为0.072 ± 0.026，紫草素1组0.177 ± 0.036， 紫草素2组0.290 ± 0.062，空白对照组0.407 ± 0.051，各组间差异有统计学意义（F = 48.895，P＜0.01），两两比较显示差异均有统计学意义（均P＜0.01）。Western印迹检测显示，小鼠表皮层中CEBPD表达水平从高至低依次为空白对照组、紫草素2组、紫草素1组、模型组，各组间CEBPD表达差异有统计学意义（F = 10.237，P＜0.05），模型组、紫草素1组和空白对照组间差异有统计学意义（均P＜0.05），而紫草素2组和空白对照组间差异无统计学意义（P＞0.05）。结论 外用紫草素可有效干预咪喹莫特诱导银屑病样小鼠模型的形成；CEBPD在银屑病样小鼠模型中表达降低，外用紫草素可显著升高CEBPD的表达。

关键词: 银屑病, 模型, 动物, 紫草素, CCAAT增强子结合蛋白质δ, 咪喹莫特

Abstract: 【Abstract】 Objective To investigate the intervention effect of topical shikonin on an imiquimod-induced psoriasis-like mouse model and its effect on expression of CCAAT enhancer binding protein δ （CEBPD）. Methods Twenty specific pathogen-free BALB/c male mice were randomly and equally divided into model group, shikonin 1 group, shikonin 2 group and blank control group by using simple random sampling. Mice in the model group, shikonin 1 group and shikonin 2 group were topically treated with 50 mg of 5% imiquimod cream every day on the shaved back to establish the psoriasis-like mouse model. After 6-hour treatment, mice in the shikonin 1 group and shikonin 2 group were treated with 0.5 ml of shikonin at concentrations of 0.576 and 5.76 g/L respectively in the modeling area for 8 consecutive days; the blank control group received no treatment. Changes in the skin lesions of these mice were observed by naked eyes every day, and evaluated by using psoriasis area severity index （PASI）; after 8-day treatment, the mice were sacrificed by cervical dislocation, the dorsal skin tissues were resected, and immunohistochemical study and Western blot analysis were performed to determine the expression of CEBPD in the mouse epidermis. Statistical analysis was carried out with SPSS 16.0 software by using one-way analysis of variance for comparisons of observation indices among different groups, as well as least significant difference-t test for multiple comparisons. Results On day 8, the mice in the model group presented with obvious erythema, scales, and infiltrative and thickened skin lesions; compared with the model group, the skin lesions were markedly improved in the shikonin 1 group and shikonin 2 group, and the improvement was more obvious in the shikonin 2 group. On day 8, the PASI score significantly differed among the blank control group, model group, shikonin 1 group and shikonin 2 group （0, 11.0 ± 1.22, 8.6 ± 0.55, 5.8 ± 1.30 points, respectively; F = 128.21, P＜0.01）, and there were significant differences between any two groups （all P < 0.01）. Immunohistochemical study showed a significant difference in the expression of CEBPD （A value） among the model group, shikonin 1 group, shikonin 2 group and blank control group （0.072 ± 0.026, 0.177 ± 0.036, 0.290 ± 0.062, 0.407 ± 0.051, respectively; F = 48.895, P < 0.01）, and there were also significant differences between any two groups （all P < 0.01）. Western blot analysis showed that the CEBPD expression in the mouse epidermis was highest in the blank control group, followed in descending order by the shikonin 2 group, shikonin 1 group and model group, and significantly differed among the above 4 groups （F = 10.237, P＜0.05）; moreover, there were significant differences in the CEBPD expression between the model group and blank control group, as well as between the shikonin 1 group and blank control group （both P＜0.05）, while no significant difference was observed between the shikonin 2 group and the blank control group （P > 0.05）. Conclusion Topical shikonin could effectively interfere with the development of imiquimod-induced psoriasis-like mouse model; CEBPD expression decreased in the psoriasis-like mouse model, and could be markedly upregulated by topical application of shikonin.

Key words: Psoriasis, Models, animal, Shikonin, CCAAT-enhancer-binding protein-delta, Imiquimod