酸化脂肪酸酯治疗特应性皮炎样模型小鼠的疗效及机制初探  

Efficacy of acidified aliphatic ester in the treatment of atopic dermatitis in mouse models and preliminary exploration of its mechanisms of action

作　　者：陈春里 闫思聿 王丹[1,2] 高丽华[1,2] 谭丽娜[1,2] 唐四元[3] 刘伟[3] 曾金容[1,2] 鲁建云[1,2] Chen Chunli;Yan Siyu;Wang Dan;Gao Lihua;Tan Lina;Tang Siyuan;Liu Wei;Zeng Jinrong;Lu Jianyun(Department of Dermatology,The Third Xiangya Hospital of Central South University,Changsha 410013,China;Medical Ozone Research Center of Central South University,Changsha 410013,China;The Xiangya Nursing School of Central South University,Changsha 410013,China)

机构地区：[1]中南大学湘雅三医院皮肤科,长沙410013 [2]中南大学医用臭氧研究中心,长沙410013 [3]中南大学湘雅护理学院,长沙410013

出　　处：《中华皮肤科杂志》2023年第9期822-831,共10页Chinese Journal of Dermatology

基　　金：湖南省自然科学基金(2022JJ80041)。

摘　　要：目的探究酸化脂肪酸酯治疗特应性皮炎(AD)样模型小鼠的疗效及初步机制。方法将6~8周龄雌性BALB/c小鼠20只,随机分成2组,空白组(5只)双耳每日涂抹无水乙醇(14.3μl/耳);模型组(15只)双耳每日涂抹卡泊三醇搽剂14.3μl/耳及20 g/L卵清蛋白25μl/耳,连续10 d,构建AD样模型小鼠;从第11天开始,将模型组小鼠随机分为AD模型组、脂肪酸酯组、酸化脂肪酸酯组,每组5只,上午各组均继续涂抹卡泊三醇搽剂和卵清蛋白维持AD样模型,下午仅脂肪酸酯组及酸化脂肪酸酯组分别外涂脂肪酸酯和酸化脂肪酸酯10μl/耳。实验期间监测小鼠体重、耳厚度、耳部皮损评分及搔抓频次变化。分别于第10、14天取小鼠耳部皮肤拭子标本进行16S rRNA微生物群落多样性分析。于第14天对耳部皮肤进行反射式共聚焦显微镜检查后处死小鼠,取耳部组织进行HE染色、肥大细胞染色及实时定量PCR(RT-qPCR)实验,取血液标本进行血清IgE检测。对满足方差齐性的数据,采用单因素方差分析,两两组间比较采用LSD-t检验。结果实验第14天,小鼠耳部皮损严重程度:AD模型组>脂肪酸酯组>酸化脂肪酸酯组>空白组,与AD模型组比较,酸化脂肪酸酯组小鼠耳厚度(F=897.50,P<0.001)、皮损评分(F=268.80,P<0.001)、搔抓频次(F=64.36,P<0.001)及表皮厚度(F=256.20,P<0.001)均显著降低,RT-qPCR提示酸化脂肪酸酯组皮损区炎症因子白细胞介素(IL)-33(F=3.38,P=0.049)、胸腺基质淋巴细胞生成素(TSLP)(F=8.70,P=0.001)、IL-4(F=41.73,P<0.001)、肿瘤坏死因子(TNF)-α(F=44.30,P<0.001)的表达及肥大细胞浸润程度(F=134.30,P<0.001)亦显著降低。微生物群落多样性分析结果提示,酸化脂肪酸酯治疗可以抑制AD样模型小鼠耳部葡萄球菌属的定植,4组之间Shannon指数和Simpson指数差异有统计学意义(F=9.00、7.92,P=0.001、0.002)。结论酸化脂肪酸酯可能通过调节免疫、抑制炎症和恢复皮肤微生态多样性改�Objective To investigate the efficacy of acidified aliphatic ester in the treatment of atopic dermatitis(AD)in mouse models,and to preliminarily explore its mechanisms of action.Methods Twenty female BALB/c mice aged 6 to 8 weeks were randomly divided into 2 groups:5 mice in the blank control group were topically treated with absolute ethanol on both ears(14.3μl per ear)every day,and 15 mice in the model group were topically treated with calcipotriol liniment(14.3μl per ear)and 20 g/L ovalbumin(25μl per ear)on both ears every day for 10 consecutive days to establish AD-like mouse models.From day 11,15 mice in the model group were randomly divided into 3 groups(5 mice in each group),including AD model group,aliphatic ester group,and acidified aliphatic ester group;in the forenoon,all the 3 groups continued to be topically treated with calcipotriol liniment and ovalbumin to maintain AD-like models;in the afternoon,the aliphatic ester group and acidified aliphatic ester group were topically treated with aliphatic ester and acidified aliphatic ester respectively(10μl per ear),and no treatment was given to the AD model group.Changes in body weight,ear thickness,ear skin lesion scores,and scratching frequency were observed.Ear skin swabs were obtained from the mice on days 10 and 14 for 16S rRNA gene-based microbial diversity tests.On day 14,mice were sacrificed after reflectance confocal microscopy examinations of the ear skin,ear tissues were resected for hematoxylin and eosin staining,mast cell staining,and real-time fluorescence-based quantitative PCR(RT-qPCR),and blood samples were collected for detection of serum IgE levels.One-way analysis of variance was used for analysis of data that met homogeneity of variance criteria,and least significant difference-t test for multiple comparisons.Results On day 14,the severity of mouse ear lesions was the highest in the AD model group,followed in turn by the aliphatic ester group,acidified aliphatic ester group,and blank control group;compared with the AD model group,t

关 键 词：皮炎 特应性 疾病模型 动物 炎症 皮肤表现 细胞因子类 微生物群落 酸化脂肪酸酯 16S rRNA 葡萄球菌属 

分 类 号：R758.2[医药卫生—皮肤病学与性病学] R-332[医药卫生—临床医学]