黄芩苷联合氟康唑对白念珠菌生物膜的抑制作用研究

中国真菌学杂志　 2015, Vol. 10 　Issue (3): 139-145.

黄芩苷联合氟康唑对白念珠菌生物膜的抑制作用研究

颜贵明^1,2, 施高翔^1,2, 邵菁^1,2, 汪天明^1,2, 夏丹^1,2, 汪长中^1,2

1. 安徽中医药大学中西医结合临床学院, 合肥 230038;
2. 安徽省中医药科学院中西医结合研究所, 合肥 230038

收稿日期:2015-03-25 出版日期:2015-06-28 发布日期:2015-06-28
通讯作者: 汪长中, E-mail:ahwcz63@sina.com E-mail:ahwcz63@sina.com
作者简介:颜贵明, 男 (汉族), 硕士, 副教授.E-mail:922-119@163.com
基金资助:
安徽省自然科学基金(1408085MH165), 安徽中医药大学校级科研项目(2014zr026)

Inhibitory effects of baicalin in combination with fluconazole against Candida albicans biofilms

YAN Gui-ming^1,2, Shi Gao-xiang^1,2, SHAO Jing^1,2, WANG Tian-ming^1,2, XIA Dan^1,2, WANG Chang-zhong^1,2

1. School of Integrated Traditional and WesternMedicine, Anhui University of Chinese Medicine, Hefei 230038, China;
2. Institute of Integrated Traditional and Western Medicine, Anhui Academy of Chinese Medicine, Hefei 230038, China

Received:2015-03-25 Online:2015-06-28 Published:2015-06-28

摘要/Abstract

摘要：

目的探讨中药有效成分黄芩苷 (baicalin,BA)联合氟康唑 (fluconazole,FLC)对白念珠菌 (Candida albicans,C.albicans)生物膜的抑制作用。方法通过棋盘法考察BA联合FLC对白念珠菌浮游菌与生物膜的部分抑菌浓度指数 (FICI);通过时间-杀菌曲线检测两药联合对白念珠菌标准株 (C.albicans SC5314)的杀菌作用;以XTT减低法和干重法检测两药联合对白念珠菌SC5314生物膜代谢及生物量的影响;采用扫描电镜 (Scanning electron microscopy,SEM)和激光共聚焦显微镜 (Confocal laser scanning microscopy,CLSM)观察两药联合对白念珠菌SC5314生物膜形态结构的影响;以水-烃法检测两药联合对白念珠菌SC5314生物膜细胞表面疏水性 (cell surface hydrophobicity,CSH)的影响;通过实时荧光定量PCR (quantitative real time PCR,qRT-PCR)检测两药联合对白念珠菌生物膜和CSH相关基因表达的影响。结果黄芩苷与氟康唑联用抗白念珠菌浮游菌的FICI介于0.28~0.75之间,对生物膜的FICI介于0.16~0.5之间,表现为协同作用;SEM和CLSM在生物膜结构上验证了两药的协同效果;两药联合可降低生物膜表面疏水性,以及使ALS1、ALS3、EAP1、SUN41和CSH1分别下调6%、51%、24%、13%和39%。结论黄芩苷具有协同氟康唑抗白念珠菌生物膜作用。

关键词: 黄芩苷, 白念珠菌, 生物膜, 氟康唑, 协同

Abstract:

Objective This study aimed to investigate the antifungal activity of baicalin alone or in combination with fluconazole against Candida albicans biofilms and to explore the related mechanism.Methods Checkerboard method was uased to study the interrelation of baicalin and fluconazole on C.albicans;Time-kill curve was used to evaluate the effect of baicalin alone and in combination with fluconazole on C.albicans;XTT reduction assay and measurement of biofilm biomass assay were performed to investigate the antibiofilm activity of baicalin alone and in combination with fluconazole;Scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) were used to observe the three-dimensional structure of C.albicans biofilms;Cellular surface hydrophobicity (CSH) assay was performed to investigate the effect of baicalin alone and in combination with fluconazole on adherence in C.albicans;Quantitative real time PCR (qRT-PCR) was performed to measure the expression of adhesion and hypal related genes.Results Synergistic effect was showed in anti-biofilm by baicalin in combination with fluconazole,and FICI was between 0.28-0.5;SEM and CLSM showed that baicalin alone and in combination with fluconazole could destruct and resulted in scant or nonexistent biofilms;And expressions of ALS1,ALS3,EAP1,SUN41,CSH1 were significantly downregulated 6%,51%,24%,13%和39% by qRT-PCR.Conclusion Synergistic effect of baicalin in combination with fluconazole on C.albicans biofilm included by influenced the expression of adhesion,CSH controlling genes,and decreased the CSH.

Key words: baicalin, C.albicans, biofilms, fluconazole, synergism

中图分类号:

R379.4

颜贵明, 施高翔, 邵菁, 汪天明, 夏丹, 汪长中. 黄芩苷联合氟康唑对白念珠菌生物膜的抑制作用研究[J]. 中国真菌学杂志, 2015, 10(3): 139-145.

YAN Gui-ming, Shi Gao-xiang, SHAO Jing, WANG Tian-ming, XIA Dan, WANG Chang-zhong. Inhibitory effects of baicalin in combination with fluconazole against Candida albicans biofilms[J]. Chinese Journal of Mycology, 2015, 10(3): 139-145.

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