Department of Dermatology, Eunpyeong St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul 03312, Korea.
Department of Dermatology, School of Medicine, Chungnam National University, Daejeon 35015, Korea.
Int J Mol Sci. 2021 Apr 27;22(9):4581. doi: 10.3390/ijms22094581.
Mesenchymal stem cell therapy (MSCT) has been shown to be a new therapeutic option for treating alopecia areata (AA). Outer root sheath cells (ORSCs) play key roles in maintaining the hair follicle structure and supporting the bulge area. In human ORSCs (hORSCs), the mechanism for this process has not been extensively studied. In this study, we aimed to examine the influence of human hematopoietic mesenchymal stem cells (hHMSCs) in the hORSCs in vitro model of AA and determine the mechanisms controlling efficacy. Interferon-gamma (IFN-γ) pretreatment was used to induce an in vitro model of AA in hORSCs. The effect of MSCT on the viability and migration of hORSCs was examined using co-cultures, the MTT assay, and migration assays. We investigated the expression of molecules related to the Wnt/β-catenin pathway, JAK/STAT pathway, and growth factors in hHMSC-treated hORSCs by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analyses. hHMSCs increased hORSC viability and migration when they were co-cultured. hHMSCs reverted IFN-γ-induced expression-including NLRP3, ASC, caspase-1, CXCL-9 through 11, IL-1β, and IL-15-and upregulated several growth factors and hair stem cell markers. hHMSCs activated several molecules in the Wnt/β-catenin signaling pathway, such as in the Wnt families, β-catenin, phosphorylated GSK-3β and cyclin D1, and suppressed the expression of DKK1 induced by IFN-γ in hORSCs. hHMSCs suppressed the phosphorylation of JAK1 to 3, STAT1, and STAT3 compared to the controls and IFN-γ-pretreated hORSCs. These results demonstrate that hHMSCs increased hORSC viability and migration in the in vitro AA model. Additionally, MSCT definitely stimulated anagen survival and hair growth in an HF organ culture model. MSCT appeared to be associated with the Wnt/β-catenin and JAK/STAT pathways in hORSCs.
间充质干细胞治疗 (MSCT) 已被证明是治疗斑秃 (AA) 的一种新的治疗选择。外根鞘细胞 (ORSCs) 在维持毛囊结构和支持隆起区域方面发挥着关键作用。在人类 ORSCs (hORSCs) 中，这一过程的机制尚未得到广泛研究。在这项研究中，我们旨在研究人造血间充质干细胞 (hHMSCs) 在体外 AA 模型中对 hORSCs 的影响，并确定控制疗效的机制。干扰素-γ (IFN-γ) 预处理用于诱导 hORSCs 体外 AA 模型。使用共培养、MTT 测定和迁移测定来检查 MSCT 对 hORSCs 活力和迁移的影响。我们通过逆转录-聚合酶链反应 (RT-PCR) 和 Western blot 分析研究了 hHMSC 处理的 hORSCs 中与 Wnt/β-catenin 通路、JAK/STAT 通路和生长因子相关分子的表达。hHMSCs 增加了共培养时 hORSC 的活力和迁移。hHMSCs 逆转了 IFN-γ诱导的表达，包括 NLRP3、ASC、caspase-1、CXCL-9 通过 11、IL-1β 和 IL-15，并上调了几种生长因子和毛干细胞标志物。hHMSCs 激活了 Wnt/β-catenin 信号通路中的几个分子，如 Wnt 家族、β-catenin、磷酸化 GSK-3β 和 cyclin D1，并抑制了 IFN-γ诱导的 hORSCs 中 DKK1 的表达。与对照组和 IFN-γ预处理的 hORSCs 相比，hHMSCs 抑制了 JAK1 到 3、STAT1 和 STAT3 的磷酸化。这些结果表明，hHMSCs 增加了体外 AA 模型中 hORSC 的活力和迁移。此外，MSCT 确实刺激了 HF 器官培养模型中的生长期存活和毛发生长。MSCT 似乎与 hORSCs 中的 Wnt/β-catenin 和 JAK/STAT 通路有关。
