ALS3/EGFR信号通路在阴道念珠菌病发病机制中的作用研究

Vulvovaginal candidiasis (VVC) remains a significant problem in women of childbearing age, but its pathogenesis is complex. From our previous research, we found that the anti-Candida function induced by pathway of EGFR maybe play an important role in the pathogenesis of VVC. On the base of previous research, in this study we will establish an EGFR gene chip. Then, depending on using gene chip which with the advantages of quantitative and high-throughput, also combining with Western blot, immunofluoresence, ELISA technology and gene silencing (siRNA) method; we make a further study on the pathway of signaling EGFR in pathogenesis of VVC in 3 parts. First, we will investigate the EGFR signal pathway in vitro models to make sure the function and characteristic of signaling pathway of EGFR of human vaginal epithelial cell challenged with Candida albicans. Second, we will utilize mutant strains constructed in the Calderone lab which is defective in ALS3 to clarify the pathogen recognition sites of the EGFR signal network and investigate the role of ALS3 in the activation of EGFR signaling pathway in human vaginal epithelial cells challenged with Candida albicans. Finally, we will investigate the role of signaling pathway of EGFR in VVC mouse models to further clarify the EGFR-mediated defensive mechanism during the course of disease. In summary, we will identify EGFR defensive mechanism in VVC from holistic, change, and host and pathogen interaction aspect. Then initially clear the function of EGFR signaling pathway in pathogenesis of VVC, and possible provide a new direction of research and clinical treatment for the pathogenesis and treatment of VVC in the future.

阴道念珠菌病（VVC）严重影响广大妇女患者的身心健康和生活质量。该病发病机制复杂，我们在前期的研究中发现阴道上皮细胞EGFR介导的通路防御作用可能在阴道念珠菌病中扮演了重要角色。我们拟在原有工作基础上，通过建立EGFR通路Oligo基因芯片，借助基因芯片技术高通量、可定量的优势，联合Western blot、免疫荧光、ELISA等技术，结合基因沉默方法，首先研究白念珠菌与阴道上皮细胞体外模型相互作用过程中发挥作用的EGFR介导信号通路及可能作用机制，然后通过改变致病菌株ALS3的性状，进行EGFR病原识别位点对信号网络所产生的影响作用研究，最后将初步结果利用动物模型可控性强的优势进行小鼠VVC体内模型验证，进一步明确EGFR介导信号通路与该疾病发生的相关性及作用；进而初步明确EGFR通路在阴道念珠菌病发病过程中的作用，为临床阴道念珠菌病的发病机制提供新的研究和治疗方向。

阴道念珠菌病（VVC）严重影响广大妇女患者的身心健康和生活质量。该病发病机制复杂，本研究从菌株自身代谢导致其致病能力变化，及白念珠菌感染阴道组织过程中EGFR通路相关因子所产生变化对疾病影响等几个方面进行研究，取得了一些有意义的发现。首先，本实验着手于研究白念珠菌自身基础生物代谢与其致病性的关系。利用分子生物学及动物模型方法从细胞形态、菌株毒力及致病力方面验证了线粒体功能缺陷对白念珠菌致病性的影响，初步明确了菌株线粒体功能缺陷会导致自身致病力降低，对今后念珠菌感染性疾病控制及抗真菌新药的研发具有一定意义；其次，本实验从非免疫因子EGFR对局部免疫应答影响的设想，利用qPCR及Western blot、免疫组化、ELISA等技术，结合siRNA及磷酸化抑制方法，验证白念珠菌感染阴道上皮细胞过程中EGFR-HER2-MAPK1(ErK)转导信号通路，并初步明确该通路在发病过程中可以诱导IL-8、IL-1等介导免疫反应的分子机制；再次，从抗原抗体结合激活免疫通路的原理出发，通过前述分子生物学方法初步明确白念珠菌粘附因子ALS3作为抗原识别位点可以对阴道上皮细胞抗白念珠菌感染过程中EGFR介导通路产生影响，为进一步探索阴道念珠菌病局部防御作用以及寻找新的治疗靶点奠定研究基础；最后，基于动物模型可以最大化的模拟人类疾病发病的优势，利用小鼠阴道念珠菌病模型对EGFR介导通路进行体内模型验证，进一步明确在阴道念珠菌病发病过程中EGFR及其介导产生的免疫反应对组织清除局部念珠菌感染的重要作用。总之，本研究通过上述几部分实验初步探索了白念珠菌自身致病性以及导致阴道感染过程中EGFR通路的作用及其机理和特征，为进一步深入、全面揭示阴道念珠菌病的发病机制奠定重要的科学研究基础，为临床阴道念珠菌病的发病机制提供新的研究和治疗方向。