S100A10蛋白在银屑病皮损的表达及对小鼠银屑病样皮炎模型的影响  

S100A10 protein expression in psoriatic lesions and its effect on psoriasis-like skin inflammation in mouse models

作　　者：付丹丹[1] 李佳林 付修宇 张芯育 胡华[1] 宋向凤 田中伟[1] Fu Dandan;Li Jialin;Fu Xiuyu;Zhang Xinyu;Hu Hua;Song Xiangfeng;Tian Zhongwei(Department of Dermatology,The First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan,China;Department of Immunology,School of Basic Medical Sciences,Xinxiang Medical University,Xinxiang 453003,Henan,China)

机构地区：[1]新乡医学院第一附属医院皮肤科,卫辉453100 [2]新乡医学院基础医学院免疫学教研室,新乡453003

出　　处：《中华皮肤科杂志》2023年第9期839-844,共6页Chinese Journal of Dermatology

基　　金：河南省医学科技攻关计划(联合共建)项目(LHGJ20190469);新乡医学院第一附属医院青年培育基金(QN-2022-A06)。

摘　　要：目的探讨S100A10蛋白在银屑病皮损的表达以及对咪喹莫特诱导小鼠银屑病样皮炎的影响。方法2020年1月至2022年6月在新乡医学院第一附属医院皮肤科通过外科手术收集28例银屑病患者皮损,同期于普通外科及眼科收集18例健康人皮肤作为对照组。通过免疫组化检测S100A10蛋白在银屑病患者皮损和健康对照皮肤的表达。分别选取10只野生型(WT)C57BL/6J雌性小鼠及10只S100A10-/-C57BL/6J雌性小鼠,建立咪喹莫特(IMQ)诱导的银屑病样皮炎小鼠模型,采用随机数字表法分为基因敲除(KO)/IMQ组、WT/IMQ组、KO/凡士林(VAS)组及WT/VAS组,每组5只,背部剃毛后,KO/IMQ及WT/IMQ组每日涂抹IMQ乳膏(62.5 mg)于背侧皮肤建立银屑病样皮炎小鼠模型,KO/VAS及WT/VAS组每日涂抹凡士林乳膏(62.5 mg),连续7 d,每日观察小鼠背部皮损情况,于第7天采用颈椎脱臼法处死,切取背部皮肤组织。每日通过银屑病皮损面积及严重程度指数(PASI)评价IMQ诱导的小鼠银屑病样皮炎,HE染色观察皮损病理表现,免疫组化染色检测小鼠皮损中S100A10、Ki-67的表达,Western印迹观察STAT3、IL-17A等细胞因子的表达,实时荧光定量PCR(qPCR)检测IL-17 mRNA的表达。采用独立样本t检验、非参数检验(U检验)、χ^(2)检验等进行统计学分析。结果免疫组化结果显示,寻常型银屑病患者皮损区S100A10蛋白表达低于健康对照皮肤组织(Z=-3.47,P<0.001)。在小鼠模型中,WT/IMQ组皮损区S100A10蛋白表达较WT/VAS组降低(t=3.64,P=0.007),KO/IMQ组Ki-67蛋白表达较WT/IMQ组升高(t=2.97,P=0.041)。KO/IMQ组小鼠较WT/IMQ组出现更严重的鳞屑、浸润等临床表现。Western印迹结果显示,KO/IMQ组p-STAT3/STAT3(t=3.27,P=0.031)、IL-17A(t=3.48,P=0.025)蛋白表达均较WT/IMQ组升高,qPCR显示,KO/IMQ组IL-17A mRNA水平也较WT/IMQ组升高(t=2.73,P=0.029)。结论S100A10蛋白在银屑病患者皮损中低表达;S100A10蛋白的缺失可能通过上调表皮STAT3磷酸化加Objective To determine S100A10 protein expression in psoriatic lesions,and to investigate its effect on psoriasis-like skin inflammation in imiquimod(IMQ)-induced mouse models.Methods From January 2020 to June 2022,skin lesions were surgically collected from 28 patients with psoriasis in the Department of Dermatology,the First Affiliated Hospital of Xinxiang Medical University;normal skin tissues were collected from 18 healthy subjects in the Department of General Surgery and Department of Ophthalmology during the same period,and served as a control group.Immunohistochemical staining was performed to determine the S100A10 protein expression in skin lesions from psoriasis patients and normal skin tissues from healthy controls.Ten wild-type(WT)C57BL/6J female mice and 10 S100A10-/-C57BL/6J female mice were selected to establish the IMQ-induced mouse models of psoriasis-like skin inflammation.Then,the mice were randomly divided into gene knock-out(KO)/IMQ group,WT/IMQ group,KO/vaseline(VAS)group,and WT/VAS group by using a random number table,and there were 5 mice in each group.The mice in the KO/IMQ group and WT/IMQ group were topically treated with IMQ cream(62.5 mg)on the shaved back daily to establish the mouse models of psoriasis-like skin inflammation,while the mice in the KO/VAS group and WT/VAS group were topically treated with vaseline cream(62.5 mg)daily,and both treatments lasted 7 consecutive days.Skin lesions on the back were observed daily.On day 7,the mice were sacrificed by cervical dislocation,and their dorsal skin tissues were excised.The IMQ-induced psoriasis-like skin inflammation was evaluated by the psoriasis area and severity index(PASI),pathological manifestations of skin lesions were observed by hematoxylin and eosin staining,the expression of S100A10 and Ki-67 in skin lesions was determined by immunohistochemical staining,the expression of STAT3,IL-17A and other cytokines was determined by Western blot analysis,and IL-17 mRNA expression was determined by real-time fluorescence-based quantit

关 键 词：银屑病 疾病模型 动物 STAT3转录因子 白细胞介素17 KI-67抗原 S100A10 白细胞介素17A 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]