钙池操纵性钙内流对特应性皮炎上皮屏障功能的影响  

Impacts of store-operated calcium entry on the function of epithelial barrier in atopic dermatitis

作　　者：曹灿 刘光辉 王畅畅 马如雪 贾欣睿 钟剑 高亚东[1] CAO Can;LIU Guanghui;WANG Changchang;MA Ruxue;JIA Xinrui;ZHONG Jian;GAO Yadong(Dept.of Allergology,Zhongnan Hospital of Wuhan University,Wuhan 430071,Hubei,China;Center for Allergy Prevention,Huazhong University of Science and Technology Union Shenzhen Hospital,Shenzhen 518052,Guangdong,China;Dept.of Dermatology,Wuhan Third Hospital,Wuhan 430060,Hubei,China)

机构地区：[1]武汉大学中南医院过敏反应科,湖北武汉430071 [2]华中科技大学协和深圳医院过敏防治中心,广东深圳518052 [3]武汉市第三医院皮肤科,湖北武汉430060

出　　处：《武汉大学学报（医学版）》2024年第4期406-412,共7页Medical Journal of Wuhan University

摘　　要：目的:探讨钙池操纵性钙内流(SOCE)及其介导的钙信号在特应性皮炎(AD)上皮屏障破坏中的作用。方法:6周龄BALB/c小鼠,采取胶带粘贴及1%DNCB致敏法建立AD模型,分为对照组、AD组、SOCE抑制剂BTP2处理的AD组(AD-BTP2)和外用糖皮质激素处理的AD组(AD-TCS)。采用组织学检查表皮厚度,通过皮水分丢失(TEWL)测定小鼠皮肤屏障功能。体外培养角质形成细胞HaCaT,分为对照组、尘螨(HDM)提取物刺激组和HDM刺激并BTP2干预组(HDM-BTP2),采用MTT法进行细胞增殖试验,RT-PCR、Western Blot法分别检测SOCE及上皮屏障相关基因的表达。FITC-右旋糖酐通透性实验评估HaCaT细胞屏障功能。结果:AD组小鼠较对照组小鼠的经皮水分丢失和表皮增厚均更为显著(P<0.0001);而BTP2处理后AD小鼠的经皮水分丢失(P<0.05)和表皮增厚(P<0.0001)进一步加重。HDM刺激可以促进HaCaT细胞的增殖,而BTP对细胞增殖没有影响。HDM刺激后HaCaT细胞SOCE相关蛋白Orai1~3,STIM1~2和SARAF的mRNA表达与对照组细胞无差异,但Orai1蛋白表达显著高于对照组(P<0.05)。HDM刺激HaCaT细胞后屏障相关闭合蛋白Occludin和兜甲蛋白Loricrin表达下调;而BTP2处理后可以恢复Occludin和增加E-cadherin的表达。HDM组HaCaT细胞的FITC-右旋糖酐通透性高于对照组细胞,而1μmol/L BTP2可以显著降低HDM诱导的FITC-右旋糖酐通透性改变。结论:在AD动物模型和细胞水平,SOCE抑制剂BTP2对上皮屏障具有相反的效应,提示SOCE在2型炎症中作用的复杂性,相关机制还需要进一步研究。Objective:To explore the impacts of store-operated calcium entry(SOCE)on the epithelial barrier of atopic dermatitis(AD).Methods:AD model in 6-week BALB/c mouse was built by tape stripping and 1%DNCB challenging and mice were divided into 4 groups:control group,AD group,AD-BTP2 group,and AD-TCS group.The effects of inhibiting SOCE in vivo were evaluated by trans-epidermal water loss(TEWL)and histology of the epidermis.HaCaT cells were cultured in vitro and cell proliferation was tested by MTT assay after HDM and BTP2 stimulation.RT-PCR and Western-Blot of SOCE and epithelial barrier-related genes in HaCaT cells proceeded for expression changes and carried out FITC-dextran assay for cell barrier function evaluation.Results:TEWL and epidermis thickness significantly increased in AD group mice than in the control group(P<0.0001).However,the BTP2-treated group showed a more significant increase in TEWL(P<0.05)and epi-dermis thickness(P<0.0001)as compared with the AD group.HDM stimulation promotes HaCaT cell proliferation,but BTP2 showed no effects.The mRNA expression levels of SOCE-related genes had no significant change after HDM stimulation in HaCaT cells.The mRNA expression of SOCE-related proteins Orai1-Orai3,STIM1,STIM1,and SARAF in HaCaT cells after HDM stimulation did not differ from those in control cells,but the protein expression of Orai1 was significantly higher(P<0.05).Occludin and Loricrin,two barrier-related proteins,decreased after HDM stimulation,and BTP2 treatment could restore the expression of Occludin and elevate E-cadherin expression.HaCaT cells in the HDM group showed higher permeability of FITC-dextran than the control group,and BTP2(1μmol/L)treatment significantly decreased the HDM-induced permeability changes.Conclusion:BTP2,the inhibitor of SOCE,shows opposite influences in the AD mouse model and HaCaT cell,which suggests the complexity of SOCE in type 2 inflammation,and further study of the underly-ing mechanisms needs to be conducted.

关 键 词：钙池操纵性钙内流 特应性皮炎 上皮屏障 

分 类 号：R758.23[医药卫生—皮肤病学与性病学]