很多临床上分离的多药耐药菌株中麦角固醇合成途径和药泵活力并无组成性的改变，真菌高度耐药的特征仅暴露在药物刺激下才诱导性地发生。因此, “药物逆境刺激而使真菌细胞激活药物逆境响应从而引起的耐药性”这条途径在临床上更需要引起关注。预实验结果表明采用钙信号通路阻断剂或钙信号蛋白或其转录因子的缺失菌株，曲霉表现出对唑类药物超敏感，表明钙信号系统参与曲霉对唑类药物的耐受性。本课题将从真菌感染的临床病人中收集曲霉耐药菌株，同时通过药物诱导和农杆菌T-DNA插入法筛选曲霉耐药突变株并采用非标记质谱定量比较耐药菌株和敏感菌株的蛋白质表达的差异；探索钙信号通路或其修饰、互作蛋白对于产生耐药的直接参与者-药物外排泵和药靶麦角固醇的合成的影响，阐明临床菌株多药耐药性与钙信号系统中功能基因的关系。探讨联合唑类药和钙信号通路阻断剂来改变耐药特征，为控制真菌的耐药性和改进现有治疗策略提供新思路。

Over the past decades, the incidence of invasive fungal infections has increased, causing serious systemic disease in immunocompromised patients, especially those caused by Aspergillus species. Up to date, azoles are widely applied and largely effective; however, they are generally fungistatic and clinically resistant isolates are emerging increasingly.The main reason for drug resistance produced by fungi is the mutation or the over-expression of multidrug efflux pumps. Thus, most of previous studies have focused to search for new drug targets or obtain new drugs to solve this problem. However, the studies about how the main player-Pleiotropic drug resistance protein was controlled by its regulators during this process are limited. In this project, based on our previous finding that synergistic activity of azoles with calcium chelators EGTA against Aspergillus nidulans and Aspergillus fumigatus, we proposed that Ca2+/calmodulin/calcineurin complex was involved in the stress-response reactions. To further completely understand the regulation mechanism of efflux pumps in Pleiotropic drug resistance activity, by using calcium chelators or inhibitors combined with activated or inactivated mutations in calcium signalling pathway, we will analysis the components of complex in this calcium signalling pathway including Ca2+-transporter,Ca2+-channels and their interaction proteins,accompanied with its transcription factors or partners during responses to azole treatment in filamentous fungi Aspergillus. Especially, We will focus on the new regulator candidates which involved in regulation of drug-efflux-pump activity by inducing calcium homeostasis. Moreover, on the basis of results from this study, we proposed the finding for this project will insight in new solution for fighting for drug resistance.