活血凉血法治疗特应性皮炎的分子免疫调控机制

Atopic dermatitis (AD) is one of the most common refractory diseases; the incidence of AD is rising year by year recently. Zhiyangrunfu Cream is an effective Traditional Chinese Medicine cream to treat AD, which is researched and developed by famous dermatological Prof. Zhang Zuozhou. The effect of the cream is to promote Huoxueliangxue. The cream has significant effect during several decades in our hospital. The mechanism of this prescription may be the regulation of Th2 immune response through blocking MAPK signaling pathways. The molecular immunological mechanism is adjusting Th1 / Th2 balance of the local tissue, restraining expression and activity of inflammatory factor and chemokine, blocking migration of T cells to reduce inflammatory response and itching, to achieve remedial effect. To verify the objectivity of the hypothesis, and discuss the improvement of dermatological symptom and inhibition of inflammatory responses, human keratinocyte cell are adopted to observe the expression and activity of inflammatory factor and chemokine and phosphorylation of MAPK signaling pathways by biological technology, such as cell culture, ELISA, real-time fluorescent quantitative quantitative PCR, western blotting and so on. The study will reveal one possible therapeutic mechanism of TCM in treating AD, clarify the effectiveness and theoretical connotation about Huoxueliangxue Method in the theory of "Treating Blood before Treating Wind", and provide new idea for inheriting experience of famous TCM doctor.

特应性皮炎(AD) 是常见难治病之一，发病率逐年上升。止痒润肤霜是著名中医皮肤外科专家张作舟教授运用活血凉血法研制的治疗AD有效验方，在我院已经应用数十年，疗效显著。为此我们推测该验方疗效的可能机制是通过MAPK信号转导通路调控Th2免疫反应，调节局部组织中Th1/Th2平衡,抑制炎性因子和趋化因子活性与表达，阻断T细胞向病灶部游走,从而减轻炎症反应和瘙痒症状，达到治疗效果。本课题采用皮肤角质形成细胞体外培养的方法，通过细胞培养、ELISA、实时荧光定量PCR、Western blotting等生物实验技术，观察炎性因子和趋化因子活性与表达、角质形成细胞内介导的MAPK通路活化等指标，揭示治疗AD的分子免疫机制，以验证假说的客观性，研究将传承挖掘名老中医"活血凉血法"治疗AD经验，揭示其理论内涵，为AD临床治疗提供新思路。

紫草具有清热、凉血、活血的功效，现代药理研究发现紫草具有很好的免疫调节、抗炎抗病毒和抗肿瘤等功能，也是著名中医皮肤外科专家张作舟教授研制的经验方止痒润肤霜的君药，作为治疗AD的我院院内制剂，已经应用数十年，疗效显著，但其在治疗过程中的作用机制尚不十分明确。本实验旨在研究紫草提取物对TNF-a和IFN-g诱导的皮肤角质形成细胞HaCaT产生Th2趋化因子的可能作用机制。我们发现提取物ZS从蛋白质、mRNA水平抑制TNF-a和IFN-g诱导下HaCaT细胞的TARC的表达，进一步研究发现，ZS明显抑制TNF-a和IFN-g诱导下的NF-kB活性；相同情况下，另一种提取物ZQ不能影响TARC表达。结果表明，紫草通过有效成分ZS发挥对皮肤角质形成细胞的免疫调节作用，抑制TNF-a和IFN-g诱导的TARC表达，并抑制NF-kB活性。由此我们推断紫草在治疗AD等皮肤病中起到免疫调节作用，揭示"活血凉血法"治疗AD经验的理论内涵。