基于热敏TRPV3通道调控角质形成细胞释放瘙痒介质探讨清热祛湿方改善特应性皮炎热觉痒化的机制研究

瘙痒是特应性皮炎（AD）最突出的临床症状，具有温热刺激及夜间加重的特征，控制瘙痒是治疗AD的重要基础。热敏瞬时受体电位（TRPV3）通道异常是介导角质形成细胞释放瘙痒介质导致热觉痒化的重要环节。中医理论认为“热微则疮痒”，清热药为止痒常用药，以清热药物为主的清热祛湿方可明显改善急性期AD皮疹及夜间瘙痒。基于前期研究，提出以下科学假说：清热祛湿方可通过TRPV3通道蛋白调控角质形成细胞释放瘙痒介质，改善温热介导的瘙痒。本项目构建TRPV3高表达的细胞和动物模型。采用免疫印迹、流式细胞、RT-qPCR，搔抓实时定量系统等检测手段，分析无害温热刺激下，TRPV3通道活化角质形成细胞释放瘙痒介质，及温热刺激下的动物行为学研究，探讨清热祛湿方是否具有调控TRPV3通道抑制角质形成细胞释放瘙痒介质的作用。期望通过本研究为中医药治疗AD的优越性提供新的理论依据。同时，对研发抗瘙痒新药提供新思路。

Pruritus is the most prominent clinical symptom of atopic dermatitis (AD), which is often aggravated by warm stimulation and at night. Controlling pruritus is an important basis for the treatment of AD. Abnormality of TRPV3 channel is an important link in the release of pruritogens from keratinocytes, leading to warmth-provoked itch. According to the theory of traditional Chinese medicine, "slight heat leads to itching". Qingre is a common method for relieving itching. Qingre Qushi recipe, which is mainly composed of Qingre herbs, can significantly improve the skin rash and itching at night in acute AD. Based on previous studies, the following scientific hypothesis was put forward: Qingre Qushi formula can regulate the release of prurituogens from keratinocytes through TRPV3 channel protein, and improve the pruritus mediated by warm heat. In this project, we constructed keratinocytes and animal models with high expression of TRPV3. Western blotting, flow cytometry, RT qPCR and scratching real-time quantitative system were used to analyze the itching signal released by keratinocytes through TRPV3 channel, and the animal behavior study under harmless warm heat stimulation, so as to explore whether Qingre Qushi formula can regulate TRPV3 channel and inhibit the release of itching mediators by keratinocytes. This study is expected to provide a new theoretical basis for the superiority of traditional Chinese medicine in the treatment of AD. At the same time, it provides new ideas for the research and development of new anti-pruritus drugs.