PDF(Pubmed)
Abstract：
BACKGROUND: Sensitive skin is hypersensitive to various external stimuli and a defective epidermal permeability barrier is an important clinical feature of sensitive skin. Claudin-5 (CLDN5) expression levels decrease in sensitive skin. This study aimed to explore the impact of CLDN5 deficiency on the permeability barrier in sensitive skin and the regulatory role of miRNAs in CLDN5 expression.
METHODS: A total of 26 patients were retrospectively enrolled, and the CLDN5 expression and permeability barrier dysfunction in vitro were assessed. Then miRNA-224-5p expression was also assessed in sensitive skin.
RESULTS: Immunofluorescence and electron microscopy revealed reduced CLDN5 expression, increased miR-224-5p expression, and disrupted intercellular junctions in sensitive skin. CLDN5 knockdown was associated with lower transepithelial electrical resistance (TEER) and Lucifer yellow penetration in keratinocytes and organotypic skin models. The RNA-seq and qRT-PCR results indicated elevated miR-224-5p expression in sensitive skin; MiR-224-5p directly interacted with the 3`UTR of CLDN5, resulting in CLDN5 deficiency in the luciferase reporter assay. Finally, miR-224-5p reduced TEER in keratinocyte cultures.
CONCLUSIONS: These results suggest that the miR-224-5p-induced reduction in CLDN5 expression leads to impaired permeability barrier function, and that miR-224-5p could be a potential therapeutic target for sensitive skin.
METHODS: A total of 26 patients were retrospectively enrolled, and the CLDN5 expression and permeability barrier dysfunction in vitro were assessed. Then miRNA-224-5p expression was also assessed in sensitive skin.
RESULTS: Immunofluorescence and electron microscopy revealed reduced CLDN5 expression, increased miR-224-5p expression, and disrupted intercellular junctions in sensitive skin. CLDN5 knockdown was associated with lower transepithelial electrical resistance (TEER) and Lucifer yellow penetration in keratinocytes and organotypic skin models. The RNA-seq and qRT-PCR results indicated elevated miR-224-5p expression in sensitive skin; MiR-224-5p directly interacted with the 3`UTR of CLDN5, resulting in CLDN5 deficiency in the luciferase reporter assay. Finally, miR-224-5p reduced TEER in keratinocyte cultures.
CONCLUSIONS: These results suggest that the miR-224-5p-induced reduction in CLDN5 expression leads to impaired permeability barrier function, and that miR-224-5p could be a potential therapeutic target for sensitive skin.
摘要：
背景：敏感性皮肤对各种外界刺激过敏，有缺陷的表皮通透性屏障是敏感性皮肤的重要临床特征。在敏感皮肤中Claudin-5(CLDN5)表达水平降低。本研究旨在探讨CLDN5缺乏对敏感皮肤通透性屏障的影响以及miRNA对CLDN5表达的调控作用。
方法：回顾性纳入26例患者，并评估了CLDN5的表达和体外通透性屏障功能障碍。然后还在敏感皮肤中评估miRNA-224-5p表达。
结果：免疫荧光和电子显微镜显示CLDN5表达减少，miR-224-5p表达增加，敏感皮肤的细胞间连接被破坏。CLDN5敲低与较低的跨上皮电阻（TEER）和在角质形成细胞和器官型皮肤模型中的路西法黄渗透有关。RNA-seq和qRT-PCR结果表明，敏感皮肤中miR-224-5p表达升高；MiR-224-5p直接与CLDN5的3'UTR相互作用，导致荧光素酶报告基因测定中的CLDN5缺陷。最后,miR-224-5p降低角质形成细胞培养物中的TEER。
结论：这些结果表明，miR-224-5p诱导的CLDN5表达减少导致通透性屏障功能受损，miR-224-5p可能是敏感皮肤的潜在治疗靶点。
方法：回顾性纳入26例患者，并评估了CLDN5的表达和体外通透性屏障功能障碍。然后还在敏感皮肤中评估miRNA-224-5p表达。
结果：免疫荧光和电子显微镜显示CLDN5表达减少，miR-224-5p表达增加，敏感皮肤的细胞间连接被破坏。CLDN5敲低与较低的跨上皮电阻（TEER）和在角质形成细胞和器官型皮肤模型中的路西法黄渗透有关。RNA-seq和qRT-PCR结果表明，敏感皮肤中miR-224-5p表达升高；MiR-224-5p直接与CLDN5的3'UTR相互作用，导致荧光素酶报告基因测定中的CLDN5缺陷。最后,miR-224-5p降低角质形成细胞培养物中的TEER。
结论：这些结果表明，miR-224-5p诱导的CLDN5表达减少导致通透性屏障功能受损，miR-224-5p可能是敏感皮肤的潜在治疗靶点。
