基于“风性开泄”理论探讨荆芥-防风药对调控TRPV1修复特应性皮炎皮肤屏障的作用及分子机制

Wind evil is the starting external cause of atopic dermatitis (AD), and its core pathogenesis is the loss of body fluid and subtle substance.The "windy leakage" characteristics of wind evil tend to cause the looseness of the skin and the exudation of the subtle substance of the body fluid. The skin barrier of AD patients is impaired, the transepidermal water loss is increased, and the expression of epidermal differentiation protein is decreased, which is consistent with the characteristics of "windy leakage". Schizonepeta and Saposhnikovia herbal pair is the most commonly used wind-expelling drug pair in AD treatment, and its mechanism may be related to barrier repairment. According to BATMAN's prediction, the drug pair can act on the ion channel TRPV1, and TRPV1 can affect the expression of filaggrin (FLG), which is closely related to the damage of the skin barrier. In this study, we tend to use NC/Nga mice to establish an animal model of AD, and to employ NHEK cells to induce the establishment of an AD epidermal cell model. The combination of in vitro and in vivo experiments will be performed to examine the effects of this herbal drug pair on FLG and related biological indicators of skin barrier. In addition, we plan to use TRPV1 and downstream p38 MAPK signaling pathway as entry points to clarify the molecular mechanism of the drug pair for repairing skin barrier by PCR, Western blot and gene overexpression. We aim to clarify the correlation with wind evil and skin barrier damage with experimental data.

风邪是特应性皮炎(AD)的始动性外因，液耗精枯是其主要病理环节。“风性开泄”，风邪可致腠理疏松、津液精微物质外泄；与现代医学表皮分化蛋白表达下降、经皮水分丢失增加，皮肤屏障受损的观点不谋而合。荆芥-防风为AD治疗中最常使用的祛风药对，根据BATMAN预测结果，该药对可作用于离子通道TRPV1，而TRPV1与丝聚蛋白(FLG)表达异常、皮肤屏障受损关系密切。荆芥-防风药对治疗AD的作用机制可能与屏障修复有关。本研究拟采用NC/Nga小鼠建立AD动物模型、利用NHEK细胞诱导AD表皮细胞模型，体内外实验相结合，检测荆芥-防风对FLG和相关皮肤屏障生物学指标的影响，验证其对皮肤屏障的修复作用。此外，我们拟以TRPV1及其下游p38 MAPK信号通路为切入点，通过PCR、Western blot和基因过表达等方法，阐明荆芥-防风药对修复皮肤屏障的分子机制，为风邪与皮肤屏障损伤的相关性提供实验依据。

背景：风邪是特应性皮炎(AD)的始动性外因， “风性开泄”，风邪可致腠理疏松、津液精微物质外泄；与现代医学表皮分化蛋白表达下降、经皮水分丢失增加，皮肤屏障受损的观点不谋而合。荆芥-防风为AD治疗中最常使用的祛风药对。.主要研究内容：本项目主要研究荆芥-防风对AD的治疗作用，并以TRPV1分子为切入点，探索其分子机制。.重要结果及关键数据如下：(1)荆芥-防风可有效缓解AD小鼠病情：荆芥-防风可缓解AD小鼠的皮损、减轻外耳皮肤厚度、减少经皮水分丢失；可缓解AD小鼠的皮损组织病理学炎症；减轻肥大细胞浸润。 (2)荆芥-防风可上调皮肤屏障相关蛋白，下调炎症因子：荆芥-防风可上调丝聚蛋白、内披蛋白等皮肤屏障相关蛋白，下调TSLP、IL-33、IL-4、TNF-α等炎症因子表达。(3)荆芥-防风可降低AD小鼠TRPV1表达：免疫组化显示，荆芥-防风可降低AD小鼠真皮层TRPV1表达；免疫荧光显示，荆芥-防风可降低AD小鼠单核巨噬细胞TRPV1表达；对CD4+T淋巴细胞TRPV1表达的影响无统计学意义。(4)荆芥-防风可调节巨噬细胞炎症模型TRPV1等炎症因子的基因表达，调节TRPV1下游信号通路分子的蛋白表达：以Raw 264.7和THP-1两种巨噬细胞系为研究对象，LPS+INF-γ刺激建立细胞炎症模型，予荆芥、防风干预后，可降低TRPV1分子及多种炎症因子表达，还可降低JAK1、p-STAT及NF-KB的表达。(5)荆芥-防风通过TRPV1分子抑制JAK信号分子表达：构建TRPV1过表达的工具细胞，予荆芥-防风干预后，TRPV1过表达细胞对JAK分子的抑制作用减弱。.科学意义：本研究证实了荆芥-防风对AD的治疗作用，以TRPV1及下游JAK1信号通路为切入点，验证了荆芥-防风治疗AD的分子机制。为AD的中医治疗提供了实验室证据。