基于“引火助阳”理论研究火针激活毛囊神经嵴干细胞治疗白癜风的分子机制

Previous investigations demonstrated that fire-acupuncture has a great potential in vitiligo treatment. However, therapeutic mechanisms underlying it remain elusive. Based on the hair follicle-initiated vitiliginous maculas recovery in clinic, the present project will be set up to explore the role of melanocyte reservoir, the hair follicular neural crest stem cell (HF-NCSCs), in the treatment. We hypothesize that fire-acupuncture treatment activate HF-NCSCs with Fire, cause the differentiation of HF-NCSCs into melanocytes, thus meet the metric of Yang in traditional Chinese medicine (TCM). Meanwhile, differentiating HF-NCSCs migrate into interfollicular epidermis, where they contribute into vitiliginous maculas recovery. To verify this hypothesis, an advanced genetic-based lineage tracing mice model for vitiligo will be generated from triple-transgenic mice. Then, upon being exposed to fire-acupuncture, HF-NCSCs will be investigated on proliferation, migration and differentiation. At last, the Wnt/beta-catenin pathway, which had been shown invovled in melanocytic differentiation, will further be analyzed systematically both in vivo and in vitro. This study will lay a foundation for clarifying the mechanisms of fire-acupuncture in vitiligo treatment and may provide some new insights into Yang theory in TCM.

既往研究表明火针治疗白癜风临床疗效确切，其治疗机制不清。本研究从临床治疗过程中出现的以毛囊为中心的“岛”状复色现象出发，提出假说：在火针治疗过程中，通过“引火”至白癜风皮损，促使皮损处的毛囊黑素前体细胞——神经嵴干细胞被激活，向黑素细胞分化，并移行至毛囊间表皮，参与皮损的复色，即助干细胞分化之“阳”。项目首先建立毛囊神经嵴干细胞遗传标记的白癜风鼠模型；然后从细胞、组织和活体水平，阐明火针对毛囊神经嵴干细胞增殖、迁移、分化的影响；最后综合分析Wnt/beta-catenin信号通路的参与机制。本研究可为阐明火针治疗白癜风的治疗机制奠定基础，同时为中医“阳气”理论提供新的科学内涵。

白癜风易诊难治。火针作为一种中医外治疗法，疗效确切但机制不清。临床采用火针治疗白癜风出现以毛囊为中心的“岛”状复色起始现象，本课题从这一现象得到启发，探究毛囊神经嵴干细胞（hair follicular neural crest stem cell, HF-NCSCs）在火针治疗白癜风中的作用。. 课题首先通过临床观察，再次确认了火针治疗白癜风的临床疗效。其次，课题引进培育了Dct-lacZ小鼠，通过标记黑素细胞特异性Dct基因，达到示踪毛囊黑素细胞的目的。接着引进培育了Krt14-Kitl小鼠，此成年小鼠表皮中含有黑素细胞，模拟人的皮肤。然后我们通过皮内注射TRP2-180多肽，诱导黑素细胞的自身免疫，建立白癜风模型。课题还针对临床火针治疗依靠操作者主观经验的缺点，研制了一种针刺密度、针刺深度、针体温度、滞针时间都精确可控的自动化火针。利用这一标准化工具，课题探索了火针治疗的最佳参数。最后，课题对火针治疗的分子机制进行了初步探索。. 结果：火针治疗白癜风和目前临床西医一线疗法（他克莫司）的疗效相当。成功建立了HF-NCSCs示踪的白癜风小鼠模型。成功研制了一种中医医疗器械——自动化火针。明确了火针治疗白癜风的分子机制是：火针通过点刺皮肤，促进Wnt的表达，启动Wnt/-catenin信号通路，导致黑素干细胞向黑素细胞分化并迁移至毛囊间表皮，最终促进白癜风皮损复色。. 本课题的研究成果，不仅丰富了火针治疗的“引火助阳”中医理论，还为火针治疗提供了科学见解和标准化工具，为下一步提高疗效打下了基础。