College of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, No.10, Poyang Lake Road, Tuanbo Xincheng West District, Jinghai District, Tianjin City, 301617, China.
Department of Dermatology, Tianjin Third Central Hospital, Tianjin City, 300170, China.
Arch Dermatol Res. 2024 Aug 23;316(8):557. doi: 10.1007/s00403-024-03281-w.
Atopic dermatitis (AD) is a chronic inflammatory skin disorder characterized by increased sensitivity to environmental allergens and irritants. Icariin, a natural compound extracted from the herb Epimedium, has been traditionally used for its potential anti-inflammatory and antioxidant properties. This study aimed to investigate the regulatory effects of icariin on AD-like symptoms and to elucidate its underlying mechanisms. The effects of icariin on TNF-α/IFN-γ-induced HaCaT cell injury were assessed using various assays, including cell counting kit-8 for cell viability, flow cytometry for reactive oxygen species (ROS) levels, and colorimetric assays for malondialdehyde (MDA) levels and superoxide dismutase (SOD) activity. In addition, the study performed enzyme-linked immunosorbent assays to assess cytokines (IL-1β, IL-6, and IL-8) and chemokines (MDC, TARC, and RANTES) levels. Flow cytometry was used to quantify apoptotic rate, while a wound-healing assay was conducted to assess cell migration. The expression of WT1 associated protein (WTAP) and serpin family B member 4 (SERPINB4) at the mRNA and protein levels was determined using qRT-PCR and western blotting, respectively. The associations between WTAP and SERPINB4 were analyzed using RNA immunoprecipitation assay and m6A RNA immunoprecipitation assay. Icariin treatment significantly mitigated TNF-α/IFN-γ-induced oxidative stress, inflammatory response, and apoptosis in HaCaT cells, while also reversing the inhibitory effect on cell migration. Icariin reduced the expression of WTAP in TNF-α/IFN-γ-stimulated HaCaT cells. Overexpression of WTAP reversed the effects of icariin in TNF-α/IFN-γ-stimulated HaCaT cells. WTAP silencing inhibited the mRNA stability of SERPINB4 through the m6A modification. SERPINB4 overexpression attenuated the effects of WTAP silencing on oxidative stress, inflammatory response, apoptosis, and migration of TNF-α/IFN-γ-stimulated HaCaT cells. Icariin treatment downregulated SERPINB4 expression by regulating WTAP in TNF-α/IFN-γ-stimulated HaCaT cells. Icariin ameliorated TNF-α/IFN-γ-induced human immortalized epidermal cell injury through the WTAP/SERPINB4 axis, highlighting the potential for targeted interventions in AD pathogenesis.
特应性皮炎（AD）是一种慢性炎症性皮肤疾病，其特征是对环境过敏原和刺激物的敏感性增加。淫羊藿素是从草药淫羊藿中提取的一种天然化合物，因其具有潜在的抗炎和抗氧化特性而被传统使用。本研究旨在探讨淫羊藿素对 AD 样症状的调节作用，并阐明其潜在机制。通过细胞计数试剂盒-8 评估淫羊藿素对 TNF-α/IFN-γ诱导的 HaCaT 细胞损伤的影响，用于细胞活力，流式细胞术用于评估活性氧 (ROS) 水平，以及比色法用于评估丙二醛 (MDA) 水平和超氧化物歧化酶 (SOD) 活性。此外，该研究进行了酶联免疫吸附测定，以评估细胞因子 (IL-1β、IL-6 和 IL-8) 和趋化因子 (MDC、TARC 和 RANTES) 水平。流式细胞术用于定量细胞凋亡率，而划痕愈合实验用于评估细胞迁移。使用 qRT-PCR 和 Western blot 分别测定 WT1 相关蛋白 (WTAP) 和丝氨酸蛋白酶家族 B 成员 4 (SERPINB4) 在 mRNA 和蛋白质水平的表达。使用 RNA 免疫沉淀测定和 m6A RNA 免疫沉淀测定分析 WTAP 和 SERPINB4 之间的关联。淫羊藿素处理显著减轻了 TNF-α/IFN-γ诱导的 HaCaT 细胞中的氧化应激、炎症反应和细胞凋亡，同时逆转了对细胞迁移的抑制作用。淫羊藿素降低了 TNF-α/IFN-γ刺激的 HaCaT 细胞中 WTAP 的表达。WTAP 的过表达逆转了淫羊藿素在 TNF-α/IFN-γ刺激的 HaCaT 细胞中的作用。WTAP 沉默通过 m6A 修饰抑制 SERPINB4 的 mRNA 稳定性。SERPINB4 的过表达减弱了 WTAP 沉默对 TNF-α/IFN-γ刺激的 HaCaT 细胞中氧化应激、炎症反应、细胞凋亡和迁移的影响。TNF-α/IFN-γ 刺激的 HaCaT 细胞中，淫羊藿素通过调节 WTAP 下调 SERPINB4 的表达。淫羊藿素通过 WTAP/SERPINB4 轴改善了 TNF-α/IFN-γ 诱导的人永生化表皮细胞损伤，突出了针对 AD 发病机制的靶向干预的潜力。
