基于Dectin-1与TSLP通路交互作用介导的抗真菌免疫应答探讨清热利湿法干预VVC的机制

Vulvovaginal candidiasis (VVC) is one of the most common vaginitis. It seriously affects the quality of life of patients. Its pathological process is closely related to the antifungal immune response. Dectin-1 is the major pattern recognition receptor on the recognition of Candida and plays an important role in mediating antifungal immunity. Our previous research has confirmed that the clear away damp-heat method could cure VVC ,which was closely related to the Dectin-1 pathway.Recent studies have found that Thymic Stromal Lymphopoietin (TSLP) is involved in the immune regulation of fungal infection, but the crosstalk of Dectin-1/TSLP pathway on the VVC has not been reported. In this research, VVC mice and vaginal epithelial cells were used, we wish to explore the mechanism of clear away damp-heat method on VVC based on the antifungal immune response mediated by the crosstalk of Dectin-1 and TSLP pathway, integrating siRNA, WB, RT-PCR, flow cytometry, Co-IP and other methods. This work is of great value to further study the mechanism of TCM on VVC and find new therapeutic targets for treatment.

外阴阴道假丝酵母菌病（VVC）是临床中最常见的阴道炎之一，严重影响患者的生活质量。VVC的病理过程与阴道局部抗真菌免疫应答密切相关。Dectin-1是识别假丝酵母菌（真菌中的一种）的主要模式识别受体，在介导抗真菌免疫应答中扮演着重要角色，我们前期研究发现在VVC中，Dectin-1通路相关因子上调，且清热利湿法治疗VVC疗效明显，与抑制Dectin-1通路有关。最新研究提示，胸腺基质淋巴细胞生成素（TSLP）亦参与了真菌感染的免疫调控，但Dectin-1和TSLP通路二者在VVC发病中的交互作用尚未见相关报道。本项目整合siRNA、WB、RT-PCR、流式细胞、Co-IP等方法技术，从细胞和动物水平阐明Dectin-1和TSLP通路之间的交互作用对抗真菌免疫应答的调控及清热利湿法的干预机制，对深入研究中医药治疗VVC的机理，寻找防治新靶点具有重要价值。

外阴阴道假丝酵母菌病（VVC）是临床最常见的阴道炎之一，严重影响着患者的生活质量。中医药在治疗VVC方面有其独特优势。Dectin-1是识别假丝酵母菌的主要模式识别受体，胸腺基质淋巴细胞生成素（TSLP）亦参与了抗真菌感染的免疫调控。Dectin-1和TSLP通路的交互作用可能介导了清热利湿法的抗真菌免疫应答。在本研究中，我们利用假丝酵母菌刺激的人阴道上皮细胞（VK2/E6E7）和假丝酵母菌感染的小鼠作为体外和体内模型，研究清热利湿法调控Dectin-1与TSLP通路交互作用介导抗真菌免疫应答治疗VVC的机制。通过本研究得出：1.清热利湿法减缓了小鼠VVC的进展，改善阴道上皮炎症破坏情况，抑制了白假丝酵母菌诱导的炎症反应。2.清热利湿法能下调Dectin-1通路、TSLP通路关键因子的mRNA和蛋白表达。3清热利湿法能抑制DC细胞的成熟、下调促炎因子TNF-α和Th2细胞因子IL-4、IL-5、IL-6、IL-13的表达。4.在研究载体中分别加入两条通路的激动剂和抑制剂，检测两条通路下游关键因子的变化，结果显示Dectin-1和TSLP通路下游因子均有改变。在研究载体中加入通路激动剂，再用清热利湿法干预后，发现DC细胞成熟标志物OX40L、促炎因子、Th2细胞因子的表达均下降。5.在两个研究载体中，用免疫共沉淀（COIP）均检测出Dectin-1和TSLP蛋白间存在交互作用。本课题结果为进一步探讨清热利湿法治疗VVC的作用机制提供实验基础。