从表皮细胞紧密连接失调研究养血解毒方对银屑病血燥证表皮屏障修复的作用机制

Tight junction between epithelial cells is a major controlling factor of skin barrier, whose dysfunction contributes to the process of psoriasis. The disrupted tight junction promotes chronic inflammatory response, which in turn amplifies the damage in psoriatic involved skin. Damage repair of epithelial barrier is one of the most effective treatments for psoriasis. The blood-dryness is the main syndrome of chronic course of patients with psoriasis. The pathogenesis of blood-dryness formed by blood deficiency caused by Yin loss with poison heat hurting. In clinical, nourishing blood and detoxification is the principle and Yang xue huo xue jie du prescription is an effective prescription. Our previous research demonstrated that decoction markedly improved the skin barrier function via strengthening claudin-1 protein expression. Along this line, we try to better our understanding of decoction via investigating the regulatory effect of decoction on the function of tight junction, as well as relative proteins expression and location in clinical psoriatic involved skin samples and animal experiments. Moreover, we employ in vitro epithelial cells model treated by inflammatory cytokines to perform gene expression microarrays, attempting to reveal the intervention effect and mechanisms of decoction on differential gene-mediated signaling pathways. Via clinical detection, animal experiments in vivo and epithelial cells research in vitro, we try to uncover the mechanisms of decoction on treating psoriasis and provide the theoretical basis for clinical application.

银屑病发病基础与皮肤屏障功能受损密切相关，表皮细胞间的紧密连接被认为是表皮屏障功能的“主要控制因子”。银屑病皮损区紧密连接的破坏与慢性炎症相互促进，形成恶性循环，维持皮损区的炎症反应。表皮屏障的修复是临床治疗的重要手段之一。血燥证是银屑病慢性病程的主要证候，病机为阴血津液亏虚、兼有毒邪，临床以养血滋阴、润燥解毒为法，养血解毒方有良好的临床疗效。前期研究证实该方明显改善表皮屏障功能，并具有恢复紧密连接蛋白（claudin-1）表达的作用。以此为线索，本项目通过临床皮损检测及动物实验，进一步明确养血解毒方对紧密连接功能、蛋白表达及定位的调节作用，并探索养血解毒方的作用环节。同时通过炎性因子刺激的表皮细胞体外模型结合表达谱芯片检测，观察养血解毒方对“差异基因→紧密连接调节通路”的干预作用，阐明养血解毒方的作用靶点。通过临床、动物、细胞多层次的研究，为养血解毒方治疗银屑病的临床应用提供生物学证据。

银屑病发病基础与皮肤屏障功能受损密切相关，表皮细胞间的紧密连接被认为是表皮屏障功能的“主要控制因子”。银屑病皮损区紧密连接的破坏与慢性炎症相互促进，形成恶性循环，维持皮损区的炎症反应。表皮屏障的修复是临床治疗的重要手段之一。血燥证是银屑病慢性病程的主要证候，病机为阴血津液亏虚、兼有毒邪，临床以养血滋阴、润燥解毒为法，养血解毒方有良好的临床疗效。本项目通过临床皮损检测及动物实验，明确养血解毒方对紧密连接功能、蛋白表达及定位的调节作用，并探索养血解毒方的作用环节。同时通过炎性因子刺激的表皮细胞体外模型结合表达谱芯片检测，观察养血解毒方对紧密连接调节通路的干预作用，阐明养血解毒方的作用靶点。临床、动物、细胞多层次的研究表明，与凉血解毒方和活血解毒方相比，养血解毒方对皮肤屏障的修复作用和紧密连接的调节作用最为显著。通过拆方和体内外研究发现，养血解毒方中养血组分的作用靶点主要体现在调节紧密连接、修复皮肤屏障，解毒组分的作用靶点主要体现在抑制T细胞活化和细胞因子分泌上。两部分共奏养血润燥，滋阴解毒的功效，从而达到治疗银屑病的目的。养血解毒方主要通过Cdc42-aPKC通路调节紧密连接，修护皮肤屏障，打破恶性循环，起到治疗银屑病的作用。该研究阐释了养血解毒方的组方原则和科学内涵，提示临床上应重视对银屑病患者皮肤屏障的保护，为临床用药提供了指导。