蛇床子及其活性成分协同氟康唑抗耐药白念珠菌作用机制研究

In recent years, the incidence of deep fungal infection and drug resistance fungi is increasing rapidly, and it’s more and more difficult to screen antifungals from chemical agents using conventional method. In this study, by combining the Traditional Chinese Medicine (TCM) theory and modern research methods, we screened several medicinal herbs from TCM antifungal prescriptions. Our results showed that Cnidium monnieri and its active ingredient osthole have significant in vitro synergistic effect with fluconazole against drug resistant Candida albicans. In TCM theory, C. monnieri has the role to dispel pathogenic wind, but there is no study focused on its synergistic effect with fluconazole against drug resistant C. albicans. The plan of this project is ① Investigate the in vivo synergistic effect of C. monnieri and its active ingredient with fluconazole against C. albicans using mouse model. ② Investigate the influence of combining these two kinds of drugs on the gene expression profile of C. albicans using gene expression microarray assay. ③ Uncover the key signaling pathways effected by combining of these two kinds of drugs using bioinformatics methods. ④ Further verify the influence of these two kinds of drugs on related signaling pathways of C. albicans by molecular biology methods. This project would provide foundation to elucidate the synergistic mechanism of these two kinds of drugs and explore new ways of combining TCM and Western medicine to find novel antifungal agents.

近年来深部真菌感染发病率越来越高，真菌的耐药程度也越来越严重，而常规的从化药中筛选发现抗真菌药物的方法变得越来越困难。本课题将传统中医理论和现代研究手段相结合，从传统中医抗菌处方中挑选抗真菌中药，通过筛选发现蛇床子及其活性成分蛇床子素具有显著的体外协同氟康唑抗耐药白念珠菌的作用。中医认为蛇床子具有祛风杀虫的作用，但目前尚未有其与氟康唑协同抗真菌的研究报道。本项目计划 ① 用小鼠真菌感染模型考察蛇床子及其活性成分与氟康唑合用是否具有体内协同抗耐药白念珠菌的作用。② 采用基因表达谱芯片的方法研究两类药物合用对白念珠菌基因表达谱的影响。③ 生物信息学分析寻找两类药物可能影响的白念珠菌关键信号通路。④ 分子生物学进一步验证两类药物合用对白念珠菌相关信号通路的影响。本项目的研究将为深入阐明两类药物的协同机制以及探索中西医结合新途径来寻找新型抗真菌药物奠定基础。

近年来深部真菌感染发病率越来越高，真菌的耐药程度也越来越严重，而常规的从化药中筛选发现抗真菌药物的方法变得越来越困难。本课题将传统中医理论和现代研究手段相结合，从传统中医抗菌处方中挑选抗真菌中药，通过筛选发现蛇床子及其活性成分蛇床子素具有显著的体外协同氟康唑抗耐药白念珠菌的作用。蛇床子为伞形科植物蛇床的干燥成熟果实，在我国已经有很长时间的临床应用历史。中医认为蛇床子具有消炎止痒的作用, 常以其与其它中药配伍用来治疗皮肤瘙痒、湿疹、脓胞疮等。现代研究表明蛇床子素是蛇床子的主要活性成分，其是天然的香豆素类化合物，具有广泛的生物和药理活性，如神经保护，成骨，免疫调节，抗肿瘤和抗炎作用。在本项目中，我们研究了蛇床子及其活性成分蛇床子素的体外抗真菌作用。棋盘式微量液基稀释法显示蛇床子素与氟康唑合用对耐药白念珠菌有显著的协同杀菌作用。而生长曲线实验也获得了类似的结果。同时，XTT实验证实了氟康唑和蛇床子素能够协同抑制白念珠菌生物被膜的形成。基因表达谱芯片结果显示，联合应用氟康唑和蛇床子素后，白念珠菌内参与氧化还原过程，能量代谢和转运的基因表达发生了显著变化。进一步的结果表明，两药联用能够使白念珠菌内源性活性氧含量明显升高。总之，这些结果表明蛇床子素与氟康唑合用能够协同抗耐药白念珠菌，而内源性活性氧的产生可能是两药发挥协同作用的重要机制。本项目的研究将为中医临床上蛇床子用于抗真菌感染的治疗提供实验室依据，同时为探索中西医结合新途径来寻找新型抗真菌药物奠定基础。