医用臭氧油通过抑制FcεRI/Syk信号通路缓解DNCB诱导的变应性接触性皮炎  

Ozonated oil alleviates dinitrochlorobenzene-induced allergic contact dermatitis via inhibiting the FcεRI/Syk signaling pathway

作　　者：付志兵 谢雅洁 曾丽月 高丽华[1,2] 喻小春[1,2] 谭丽娜 周璐[1,2] 曾金容[1,2] 鲁建云 FU Zhibing;XIE Yajie;ZENG Liyue;GAO Lihua;YU Xiaochun;TAN Lina;ZHOU Lu;ZENG Jinrong;LU Jianyun(Department of Dermatology,Third Xiangya Hospital,Central South University,Changsha 410013;Medical Ozone Research Center of Central South University,Changsha 410013;Xiangya School of Medicine,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]中南大学医学臭氧研究中心,长沙410013 [3]中南大学湘雅医学院,长沙410013

出　　处：《中南大学学报（医学版）》2023年第1期1-14,共14页Journal of Central South University ：Medical Science

基　　金：supported by the National Natural Science Foundation(81903219);the Natural Science Foundation of Hunan Province(2018JJ6094),China.

摘　　要：目的:臭氧被广泛用于治疗过敏性皮肤病,如湿疹、特应性皮炎和接触性皮炎。然而其具体的机制仍然不清楚。本研究旨在探讨医用臭氧油对2,4-二硝基氯苯(2,4-dinitrochlorobenzene,DNCB)诱导的变应性接触性皮炎(allergic contact dermatitis,ACD)的治疗作用及其机制。方法:除空白对照组外,其余小鼠均用DNCB处理。采用DNCB建立ACD样小鼠模型,随机分为模型组、基础油组、医用臭氧油组、FcεRI过表达质粒(FcεRI-OE)组和FcεRI空质粒(FcεRI-NC)组;基础油和医用臭氧油组分别采用同等剂量基础油和医用臭氧油进行处理,FcεRI-OE组和FcεRI-NC组分别皮内注射25μg FcεRI过表达质粒和空质粒。记录皮损每日的变化,并使用反射共聚焦显微镜(reflectance confocal microscope,RCM)评估皮损厚度和炎症改变,同时对皮损组织进行苏木精-伊红(hematoxylineosin,HE)染色、实时聚合酶链反应(real-time PCR)、RNA测序(RNA-sequencing,RNA-seq)和免疫组织化学的检测及分析。结果:医用臭氧油显著减轻了DNCB诱导的ACD样皮炎,并降低了IFN-γ、IL-17A、IL-1β、TNF-α和其他相关炎症因子的表达(均P<0.05)。RNA-seq分析显示医用臭氧油显著抑制了DNCB诱导的FcεRI/Syk信号通路的激活,后续通过real-time PCR和免疫组织化学方法得到证实(均P<0.05)。与臭氧油组和FcεRI-NC组相比,FcεRI-OE组的IFN-γ、IL-17A、IL-1β、IL-6、TNF-α和其他炎症基因的mRNA表达水平明显升高(均P<0.05),FcεRI-OE组FcεRI和Syk的mRNA和蛋白质表达水平也明显升高(均P<0.05)。结论:医用臭氧油通过抑制FcεRI/Syk信号通路显著改善ACD样皮炎,减轻DNCB诱导的ACD样皮炎。Objective:Ozone is widely applied to treat allergic skin diseases such as eczema,atopic dermatitis,and contact dermatitis.However,the specific mechanism remains unclear.This study aims to investigate the effects of ozonated oil on treating 2,4-dinitrochlorobenzene(DNCB)-induced allergic contact dermatitis(ACD)and the underling mechanisms.Methods:Besides the blank control(Ctrl)group,all other mice were treated with DNCB to establish an ACD-like mouse model and were randomized into following groups:a model group,a basal oil group,an ozonated oil group,a FcεRI-overexpressed plasmid(FcεRI-OE)group,and a FcεRI empty plasmid(FcεRI-NC)group.The basal oil group and the ozonated oil group were treated with basal oil and ozonated oil,respectively.The FcεRI-OE group and the FcεRI-NC group were intradermally injected 25μg FcεRI overexpression plasmid and 25μg FcεRI empty plasmid when treating with ozonated oil,respectively.We recorded skin lesions daily and used reflectance confocal microscope(RCM)to evaluate thickness and inflammatory changes of skin lesions.Hematoxylin-eosin(HE)staining,real-time PCR,RNA-sequencing(RNA-seq),and immunohistochemistry were performed to detct and analyze the skin lesions.Results:Ozonated oil significantly alleviated DNCB-induced ACD-like dermatitis and reduced the expressions of IFN-γ,IL-17A,IL-1β,TNF-α,and other related inflammatory factors(all P<0.05).RNA-seq analysis revealed that ozonated oil significantly inhibited the activation of the DNCB-induced FcεRI/Syk signaling pathway,confirmed by real-time PCR and immunohistochemistry(all P<0.05).Compared with the ozonated oil group and the FcεRI-NC group,the mRNA expression levels of IFN-γ,IL-17A,IL-1β,IL-6,TNF-α,and other inflammatory genes in the FcεRI-OE group were significantly increased(all P<0.05),and the mRNA and protein expression levels of FcεRI and Syk were significantly elevated in the FcεRI-OE group as well(all P<0.05).Conclusion:Ozonated oil significantly improves ACD-like dermatitis and alleviated DNCB-induced

关 键 词：医用臭氧油 2 4二硝基氯苯 变应性接触性皮炎 FcεRI/Syk信号通路 

分 类 号：R758.22[医药卫生—皮肤病学与性病学]