不同细胞因子联合咪喹莫特诱导银屑病样皮损大鼠模型的比较研究  

A comparative study on rat models of psoriasis-like lesions induced by different cytokines combined with imiquimod

作　　者：王楠楠 蔡婷婷[1] 刘霞[1] 朱婉萍[1] Wang Nannan;Cai Tingting;Liu Xia;Zhu Wanping(Zhejiang Academy of Traditional Chinese Medicine,Zhejiang Provincial Key Laboratory of Traditional Chinese Medicine for Pharmacodynamic Material Basis Research of Chinese Medicine,Key Laboratory of Research and Development of Chinese Medicine of Zhejiang Province,Hangzhou 310007,China)

机构地区：[1]浙江省中医药研究院,浙江省中药药效物质基础研究中医药重点实验室,浙江省中药新药研发重点实验室,杭州310007

出　　处：《中华皮肤科杂志》2023年第12期1146-1153,共8页Chinese Journal of Dermatology

基　　金：浙江省基础公益研究计划项目(LGD21H310001)。

摘　　要：目的探讨白细胞介素(IL)23/Th17轴的相关细胞因子联合咪喹莫特诱导银屑病样皮炎大鼠模型的可行性。方法将110只Wistar大鼠按体重相对均匀分为正常对照组、咪喹莫特单用组、咪喹莫特联用干扰素(IFN)α2a(18万、6万、2万IU/kg)组、咪喹莫特联用肿瘤坏死因子(TNF)α(4.5万、1.5万、0.5万IU/kg)组及咪喹莫特联用IL-2(9万、3万、1万IU/kg)组,每组10只。将大鼠背部中央区域(2 cm×2 cm)的毛发脱去后,咪喹莫特单用组大鼠背部按照20 mg/cm^(2)均匀涂抹5%咪喹莫特乳膏;咪喹莫特联用各细胞因子组大鼠背部按相同剂量涂抹5%咪喹莫特乳膏,并于涂抹15 min后,腹腔注射相应剂量细胞因子注射液,每日1次,连续10 d。其间每日对大鼠背部皮肤进行银屑病皮损面积与严重程度指数(PASI)评分。第10天麻醉后,收集血清,采用酶联免疫吸附试验(ELISA)检测各组血清中IL-17A、TNF-α、IL-23、IFN-α及IL-1β水平;大鼠处死后,取部分皮损进行组织病理学观察并进行Baker评分,采用免疫组化法检测部分皮损中CD4、CD8的表达情况。多组间比较采用单因素方差分析,组间两两比较采用LSD-t检验。方差不齐的数据,采用Kruskal-Wallis H检验。结果造模后第3天开始,咪喹莫特单用组及联用各细胞因子组大鼠皮肤均出现红斑、鳞屑、表皮增厚等银屑病临床特征,且单用组PASI评分于第7天达峰,各联用组第6天达峰。第10天时,咪喹莫特单用及联用组皮肤组织病理学上均可见过度角化、角化不全、棘层肥厚、颗粒层变薄或消失等不同程度的银屑病病理特征。正常对照组、咪喹莫特单用及各联用组PASI评分、Baker评分差异均有统计学意义(H=43.33、F=42.15,均P<0.001)。咪喹莫特联用IFN-α2a(18万IU/kg)组及联用IL-2(9万IU/kg)组PASI评分(9.4±1.1、8.8±0.6)均高于咪喹莫特单用组(7.5±1.1,P值分别为0.002、0.030);咪喹莫特联用IFN-α2a(18、6万IU/kg)组、联用TNF-α(Objective To investigate the feasibility of construction of rat models of psoriasis-like lesions by using interleukin(IL)-23/T-helper 17(Th17)axis-related cytokines combined with imiquimod.Methods A total of 110 Wistar rats were randomly divided into normal control group,imiquimod alone group,imiquimod combined with interferon(IFN)-α2a(180000,60000,20000 IU/kg)groups,imiquimod combined with tumor necrosis factor(TNF)-α(45000,15000,5000 IU/kg)groups,and imiquimod combined with IL-2(90000,30000,10000 IU/kg)groups,and there were 10 rats in each group.After hair removal from the central area(2 cm×2 cm)of the rat back,rats in the imiquimod alone group were topically treated with imiquimod 5%cream at a dose of 20 mg/cm^(2) on the shaved back;rats in the imiquimod combined with different cytokine groups were treated with topical imiquimod 5%cream at the same dose on the shaved back for 15 minutes followed by intraperitoneal injections of cytokines at corresponding doses once a day for 10 consecutive days.During the treatment,skin lesions on the rat back were evaluated by using the psoriasis area and severity index(PASI)scores every day.On day 10,serum samples were collected from the rats after anesthesia,and enzyme-linked immunosorbent assay(ELISA)was performed to detect levels of IL-17A,TNF-α,IL-23,IFN-αand IL-1βin the serum samples in each group;then,the rats were sacrificed,lesional skin tissues on the rat back were taken for histopathological examinations and evaluated by Baker scores;an immunohistochemical study was conducted to determine the expression of CD4 and CD8 in some skin lesions.One-way analysis of variance was used for comparisons among multiple groups,and least significant difference(LSD)-t test for multiple comparisons;for data with heterogeneous variance,the Kruskal-Wallis H test was used.Results On day 3 after molding,the rats in the imiquimod alone group and combination groups gradually presented with psoriasis-like skin manifestations,such as erythema,scales and epidermal thickening;the PASI

关 键 词：银屑病 疾病模型 动物 干扰素Α 白细胞介素2 肿瘤坏死因子α 白细胞介素23 咪喹莫特 

分 类 号：R758.63[医药卫生—皮肤病学与性病学] R-332[医药卫生—临床医学]