以pH通路为靶标研究白头翁汤正丁醇提取物对白念珠菌VVC临床株形态转化及生物膜形成的抑制作用及机制

Vulvovaginal candidiasis (VVC) is a common disease to trouble women. The treatments of Azoles are easy to come into resistance, and the high recurrence rate of VVC is closely associated with yeast-hypha transition and biofilm formation of Candida albicans (Ca). It was demonstrated the pH signal pathway was involved in the yeast-hypha transition and biofilm formation of Ca. Clinically, BaiTouWeng Decoction was favourable to treat VVC, however, the underlying mechanism remained unclear. Rescently, it was found that the butyl alcohol extract of BaiTouWeng Decoction (BAEB) could effectively inhibit hypha growth and biofilm formation of Ca in vitro and in rat vagina. Therefore, it is assumed that BAEB could treat VVC through pH signal pathway. To clarify the mechanism, the VVC models in vitro and in vivo will be established, and the effect of BAEB on the yeast-hypha transition and biofilm formation of Ca will be investigated by XTT、SEM、CLSM、qRT-PCR and gene knockout from molecule, cell and animal leves. The expressions of genes involved in pH pathway, downstream hypha-specific genes and biofilm-related genes will be the emphasis to illuminate the relevant mechanism, in order to provide solid experimental basis for the treatment of BaiTouWeng Decoction and the definition of underlying target.

外阴阴道念珠菌病（VVC）是困扰多数女性的一种常见病，唑类药治疗易产生耐药性且复发率高，其发病与白念珠菌（Ca）酵母-菌丝形态转化与生物膜形成密切相关。研究证实pH信号通路参与了Ca的形态转化与生物膜形成。临床上白头翁汤治疗VVC具有良好的疗效，但作用机制不详。课题组对该复方进一步萃取，发现其正丁醇提取物（BAEB）能显著抑制体外及小鼠阴道内Ca菌丝及生物膜形成，并明显下调pH通路中形态转化基因。因此，我们提出假设：BAEB抗VVC可能通过pH通路发挥干预效应。为阐明其作用机制，拟构建体外及小鼠VVC模型，运用 XTT法、SEM、CLSM、qRT-PCR、菌株基因敲除等现代微生物学、分子生物学技术从不同层面探讨BAEB对Ca形态转化与生物膜的作用，并着重从pH通路基因、下游菌丝特异性基因与生物膜相关基因等角度阐明其作用机制，以期为白头翁汤治疗VVC以及明确其可能作用靶点提供可靠的实验依据。

外阴阴道念珠菌病（VVC）是困扰多数女性的一种常见病，唑类药治疗易产生耐药性且复发率高，其发病与白念珠菌（Ca）酵母-菌丝形态转化与生物膜形成密切相关。研究证实pH信号通路参与了Ca的形态转化与生物膜形成。临床上白头翁汤治疗VVC具有良好的疗效，但作用机制不详。课题组对该复方进一步萃取，发现其正丁醇提取物（BAEB）能显著抑制体外及小鼠阴道内Ca菌丝及生物膜形成，并明显下调pH通路中形态转化基因。因此，我们认为BAEB抗VVC可能通过pH通路发挥干预效应。本项目构建体外及小鼠VVC模型，运用XTT法、SEM、CLSM、qRT-PCR等现代微生物学、分子生物学技术从不同层面探讨BAEB对Ca形态转化与生物膜的作用，并着重从pH通路基因、下游菌丝特异性基因与生物膜相关基因等角度阐明其作用机制。结果发现：1，BAEB对VVC小鼠具有显著的治疗作用；2，BAEB可以通过pH信号通路抑制白念珠菌生物膜的形成；3，BAEB可以通过pH信号通路抑制白念珠菌的黏附作用；4，BAEB对白念珠菌VVC临床株体外生物膜形成具有抑制作用；5，BAEB对白念珠菌细胞壁具有抑制作用；6，BAEB对白念珠菌细胞膜具有抑制作用。本项目的完成为中药复方白头翁汤治疗VVC以及明确其可能作用靶点提供了可靠的科学依据。