Peng Cheng, Zhang Hualin, Yang Jishun, Xu Jianguo, Guan Shikui, Xia Jianjun, Zhu Quangang, You Benming, Zhu Yu, Hu Jinhong, Liu Jiyong
Department of Health Management, Beidaihe Rest and Recuperation Center, Joint Logistic Support Force of the Chinese People's Liberation Army, Qinhuangdao, Hebei, China.
Department of Pharmacy, 81 Group Army Hospital, PLA, Zhangjiakou, Hebei, China.
Postepy Dermatol Alergol. 2022 Jun;39(3):565-579. doi: 10.5114/ada.2022.117739. Epub 2022 Jul 14.
As a key chemotactic factor during Eos recruitment on the allergic inflammation site, eotaxin is regarded as one of the important therapeutic targets.
To address the expression and regulation mechanism of eotaxin, which constitutes an important procedure in skin allergic disease and a target for drug therapy.
An allergic contact dermatitis (ACD) model of mouse was established. Immunohistochemical method (ICH) and flow cytometry method (FCM) were used to determine the amounts of CD4+ and CD8+ T cells and their ratios. The eotaxin mRNA and protein were evaluated by real-time PCR, ICH and western-blotting method. Nuclear factor-κB (NF-κB) nuclear translocation and STAT6 phosphorylation were studied by EMSA and western-blotting methods.
We confirmed that both CD4+ and CD8+ T cells in mouse blood and tissue increased during the allergic process, FBs was the main source for eotaxin under the allergic condition. Both TNF-α and IL-4 showed synergic effects on the up-regulation of eotaxin mRNA and protein in KC and FBs. Eotaxin can be expressed via NF-κB and STAT6 transcription after KC and FBs were stimulated by TNF-α and IL-4.
The obvious up-regulation of eotaxin expression in skin tissue of the mouse ACD model was confirmed, the exact expression site and dynamic process was determined both in vivo and in vitro. The eotaxin expression ability of FBs outperformed that of KC, and eotaxin expression can be regulated by TNF-α and IL-4 via NF-κB and STAT6. The overall findings may pave the way for discovering targets for new drugs and new therapeutic drugs for treating allergic diseases.
作为嗜酸性粒细胞(Eos)在过敏性炎症部位募集过程中的关键趋化因子,嗜酸性粒细胞趋化因子(eotaxin)被视为重要的治疗靶点之一。
探讨eotaxin的表达及调控机制,这是皮肤过敏性疾病的重要过程及药物治疗靶点。
建立小鼠过敏性接触性皮炎(ACD)模型。采用免疫组织化学方法(ICH)和流式细胞术(FCM)检测CD4 +和CD8 + T细胞数量及其比例。通过实时PCR、ICH和蛋白质印迹法评估eotaxin mRNA和蛋白水平。采用电泳迁移率变动分析(EMSA)和蛋白质印迹法研究核因子-κB(NF-κB)核转位和信号转导及转录激活因子6(STAT6)磷酸化情况。
我们证实,在过敏过程中小鼠血液和组织中的CD4 +和CD8 + T细胞均增加,成纤维细胞(FBs)是过敏条件下eotaxin的主要来源。肿瘤坏死因子-α(TNF-α)和白细胞介素-4(IL-4)对角质形成细胞(KC)和成纤维细胞中eotaxin mRNA和蛋白的上调均显示协同作用。在KC和成纤维细胞受到TNF-α和IL-4刺激后,eotaxin可通过NF-κB和STAT6转录表达。
证实小鼠ACD模型皮肤组织中eotaxin表达明显上调,确定了其在体内和体外的确切表达部位及动态过程。FBs的eotaxin表达能力优于KC,且eotaxin表达可由TNF-α和IL-4通过NF-κB和STAT6进行调控。总体研究结果可能为发现治疗过敏性疾病的新药靶点和新型治疗药物铺平道路。