背景Hailey-Hailey病(HHD)是一种罕见的,常染色体显性,以表皮棘层松解症为特征的遗传性皮肤病。HHD相关基因ATPase钙转运2C型成员1(ATP2C1)编码蛋白质分泌途径Ca2ATPase1(SPCA1),在HHD发病机制中起关键作用。目的我们旨在研究ATP2C1敲低对HHD中模拟棘层松解的角质形成细胞的影响。方法采用免疫组织化学(IHC)法检测SPCA1、P-cofilin等细胞骨架蛋白和紧密连接蛋白的水平,F-肌动蛋白,Claudins,occludin,在HHD患者的皮肤活检中和zonula闭塞1。随后,使用Western印迹和免疫荧光分析了这些蛋白在培养的ATP2C1敲低角质形成细胞中的表达。此外,我们评估了扩散,凋亡,和ATP2C1敲除的角质形成细胞中的细胞内Ca2浓度。结果结果显示这些蛋白质(SPCA1,P-cofilin,F-肌动蛋白,Claudins,闭塞,和小带闭塞1)在HHD皮肤病变中。此外,它们在用ATP2C1短发夹RNA转染的人角质形成细胞中的水平降低,伴有形态棘皮松解。此外,角质形成细胞的增殖和凋亡,以及这些细胞中的细胞内钙浓度,没有受到影响。局限性这项研究的局限性是缺乏动物实验以及未能探索骨骼和紧密连接蛋白之间的关系。结论本研究表明,ATP2C1抑制导致角质形成细胞中细胞骨架和紧密连接蛋白的异常水平。因此,角质形成细胞可以模拟HHD样棘皮松解,并作为体外模型,帮助制定针对HHD的治疗策略。
Background Hailey-Hailey disease (HHD) is a rare, autosomal dominant, hereditary skin disorder characterised by epidermal acantholysis. The HHD-associated gene ATPase calcium-transporting type 2C member 1 (ATP2C1) encodes the protein secretory pathway Ca2+ ATPase1 (SPCA1), playing a critical role in HHD pathogenesis. Aims We aimed to investigate the effect of ATP2C1 knockdown on keratinocytes that mimicked acantholysis in HHD. Methods Immunohistochemistry (IHC) was employed to evaluate the levels of cytoskeletal and tight junction proteins such as SPCA1, P-cofilin, F-actin, claudins, occludin, and zonula occludens 1 in the skin biopsies of patients with HHD. Subsequently, the expression of these proteins in cultured ATP2C1 knockdown keratinocytes was analysed using Western blotting and immunofluorescence. Furthermore, we assessed the proliferation, apoptosis, and intracellular Ca2+ concentrations in the ATP2C1-knocked keratinocytes. Results The results showed decreased levels of these proteins (SPCA1, P-cofilin, F-actin, claudins, occluding, and zonula occludens 1) in HHD skin lesions. Moreover, their levels decreased in human keratinocytes transfected with ATP2C1 short hairpin RNA, accompanied by morphological acantholysis. Furthermore, the proliferation and apoptosis of the keratinocytes, as well as intracellular calcium concentrations in these cells, were not affected. Limitations The limitations of this study are the absence of animal experiments and the failure to explore the relationship between skeletal and tight junction proteins. Conclusion The present study indicated that ATP2C1 inhibition led to abnormal levels of the cytoskeletal and tight junction proteins in the keratinocytes. Therefore, keratinocytes can mimic HHD-like acantholysis and serve as an in vitro model, helping develop treatment strategies against HHD.