芍药甘草汤通过调控Notch信号通路影响Th17/Treg失衡治疗重症斑秃的机制研究

Alopecia areata is a T cell-induced autoimmune disease. Severe alopecia areata, defined as 50% area of hair loss, is not a self-limited disease. The dysfunction of T cells is an important cause for the onset of alopecia areata. Current studies demonstrate that Th17/regulatory T cell (Treg) imbalance is closely associated with the onset of alopecia areata. Our previous clinical trials showed that Shaoyao-Gancao Decoction, for the liver constraint and spleen deficiency syndrome, was effective in treating alopecia areata. We also discovered that patients' lesions before and after treatment or the expressions of peripheral blood were associated with the Notch signaling pathway, IL-17A and Foxp3, which indicated the mechanism of Shaoyao-Gancao Decoction for alopecia areata was possibly by regulating Th17/Treg imbalance through the Notch signaling pathway. We will use larger sample size in this study, from tissue, protein and genes level to explore the related protein of Notch signaling pathway and the regulation of Th17/Treg-related cytokines in patients with alopecia areata treated by Shaoyao-Gancao Decoction. We will build the cellular models of Notch signaling pathway inhibition by isolating and cultivating the CD4+ T cells of patients with severe alopecia areata. We also will transplant the lesion tissues of patients with alopecia areata to SCID rats to build models. By doing these studies, we aim at exploring the mechanism of the effects of Shaoyao-Gancao Decoction, and providing objective evidence for Shaoyao-Gancao Decoction for the treatment of Severe Alopecia Areata.

斑秃是一种T细胞介导的自身免疫性疾病，50%以上的毛发脱落称为重症斑秃，很难自愈。T细胞功能异常是斑秃发病重要原因，最新的研究显示Th17 /Treg失衡与斑秃发病密切相关。我们从肝郁脾虚的角度，采用芍药甘草汤治疗重症斑秃取得确切的临床疗效，并发现Notch信号分子、IL-17A和Foxp3在患者治疗前后的皮损或外周血表达有相关性，提示芍药甘草汤的治疗机理可能是通过Notch信号通路调控Th17/Treg失衡。本研究拟扩大临床样本，从组织、蛋白和基因水平探讨芍药甘草汤对重症斑秃患者Notch信号通路相关蛋白及Th17/Treg相关细胞因子的调节作用；分离、培养重症斑秃病患者CD4+T淋巴细胞，建立Notch信号通路抑制的细胞学模型，移植斑秃患者皮损组织到SCID小鼠建立斑秃模型，来探讨芍药甘草汤作用机制，为该方治疗重症斑秃提供客观证据。

本项目验证活动期的斑秃皮损毛囊周围确实存在明显的CD4+T细胞浸润，Th17/Treg细胞存在明显的失衡，Notch1受体的上调参与了斑秃的发病，IL-23可能为斑秃致病因子。动物实验方面，探索了小鼠斑秃造模的新方法，为后续研究提供新思路。中药“芍药甘草颗粒”可以明显改善重症斑禿患者的主观症状、白色或黑色毳毛生长情况、毛发镜征象情况、头皮炎症分级的严重程度。并且芍药甘草颗粒可以抑制N、CD3+、CD3+ CD4+和CD3+ CD8+，并下调其比值。芍药甘草颗粒可以有效调节IL-4、IL-6、IL-21、IL-22、IL-23和IL-9的水平。