咪喹莫特两种给药方式构建斑秃动物模型的稳定性研究  

Stability research on the construction of a mouse model for alopecia areata using two modeling methods with imiquimod

作　　者：郑玲玲 李旺廷 张璐璐 杜洋 靳福星 周旋 翁琰 王婧雯 ZHENG Lingling;LI Wangting;ZHANG Lulu;DU Yang;JIN Fuxing;ZHOU Xuan;WENG Yan;WANG Jingwen(Department of Pharmacy,Xijing Hospital,Air Force Medical University,Xi'an 710032,China;College of Pharmacy,Shaanxi University of Chinese Medicine,Xianyang 712046,China)

机构地区：[1]空军军医大学西京医院药剂科,陕西西安710032 [2]陕西中医药大学药学院,陕西咸阳712046

出　　处：《空军军医大学学报》2025年第4期514-520,共7页Journal of Air Force Medical University

基　　金：陕西省卫生健康新药研发与精准用药科研创新平台项目(2023PT-10);中国药学会医院药学专业委员会人才专项(CPA-Z05-ZC-2022-003)。

摘　　要：目的通过比较咪喹莫特两种不同造模方式构建的斑秃(AA)小鼠模型,探索稳定的AA动物模型的构建方法。方法48只5~7周龄C57BL/6雄性小鼠随机分成4组。模型组1和空白对照组1:1次/d、同一时间在小鼠背部涂抹0.2 g咪喹莫特乳膏或凡士林,连续7 d;模型组2和空白对照组2:3次/周、同一时间在小鼠背部涂抹0.2 g咪喹莫特乳膏或凡士林,持续35 d。两种造模方式给药结束后均观察1周。实验中观测小鼠毛发宏观变化,持续监测体质量生长状态变化。给药结束与观察期后分别采集脱毛皮损部位,通过ELISA检测皮损部位IFN-γ浓度表达情况,HE染色观察皮肤毛囊数量变化情况,免疫组化检测脱毛区域附近毛囊CD4^(+)、CD8^(+)T细胞浸润情况。结果咪喹莫特两种给药方式造模后,模型组1小鼠于第3日出现涂抹部位毛发脱落,7 d后脱落完全;造模后潮红的脱毛部位在观察期第5日转呈青色,第7日开始有微小毛发生长,毛囊表现为造模7 d后萎缩并于观察期内再度恢复生长;与空白对照组1相比,CD4^(+)、CD8^(+)T细胞在造模结束时出现明显浸润且IFN-γ表达显著(P<0.01),但观察期内CD4^(+)、CD8^(+)T细胞浸润减少且IFN-γ表达明显下降;模型组2在造模第35日明显引起宏观毛发脱落,毛发镜可见脱毛部位皮肤光滑;组织病理学显示,与空白对照组2相比毛囊数量大量减少,CD4^(+)、CD8^(+)T细胞浸润增加明显,IFN-γ显著性表达(P<0.01),且相关各项指标在观察期内未有明显变化。结论外用咪喹莫特每周3次、持续35 d建立的C57BL/6 AA模型小鼠,模型持续时间更长,与AA患者发病机制临床吻合度更高,是一种相对稳定、理想的研究AA疾病的动物模型。Objective To compare the mouse models of alopecia areata(AA)constructed by two different methods of imiquimod,and explore the stable method for establishing an AA animal model.Methods A total of 48 male C57BL/6 mice aged 57 weeks were randomly divided into four groups.Model group 1 and Control group 1 received 0.2 g either imiquimod or Vaseline on the back of mice 1 time/day at the same time for seven consecutive days.Model group 2 and Control group 2 received 0.2 g of imiquimod or Vaseline on the same area 3 times/week at the same time for 35 d.After the completion of administration,both groups of mice were observed for a week.During the experiment,macroscopic changes in mouse hair were observed,and changes in body mass growth status of mice were continuously monitored.The depilated skin lesions were collected at the end of administration and after the observation period.The expression of IFN-γconcentration in the skin lesions was detected using ELISA,and the changes in the number of hair follicles were observed through HE staining.Additionally,immunohistochemistry was utilized to detect the infiltration of CD4^(+)and CD8^(+)T cells in hair follicles near the hair removal area.Results After two modeling approaches of imiquimod,the mice in Model group 1 exhibited alopecia on their dorsal and cervical regions on the third day,which progressed to complete hair loss after 7 d.Following modeling,a cyan hue appeared on the depilated area on the fifth day of observation,and micro-hair growth commenced on the seventh day.On the seventh day of modeling period,hair follicle atrophy followed by subsequent regrowth during the observation period was observed.In comparison to Control group 1,significant infiltration of CD4^(+)and CD8^(+)T cells along with IFN-γexpression were evident at the end of modeling(P<0.01),but CD4^(+)and CD8^(+)T cell infiltration and IFN-γexpression decreased significantly during the observation period.In Model group 2,macroscopic hair loss was noted on the thirty-fifth day after modeling and micr

关 键 词：斑秃 动物模型稳定性 咪喹莫特 自身免疫性疾病 

分 类 号：R758.71[医药卫生—皮肤病学与性病学] R-332[医药卫生—临床医学]