龙胆泻肝汤氯仿提取物对白念珠菌VVC临床株菌丝抑制作用研究

中国真菌学杂志　 2016, Vol. 11 　Issue (6): 341-347.

龙胆泻肝汤氯仿提取物对白念珠菌VVC临床株菌丝抑制作用研究

王霞^1,2, 吴大强^1,2, 施高翔^1,2, 段强军^1,2, 邵菁^1,2, 汪天明^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:2016-08-08 出版日期:2016-12-28 发布日期:2016-12-28
通讯作者: 汪长中,E-mail:ahwcz63@sina.com E-mail:ahwcz63@sina.com
作者简介:王霞,女(汉族),硕士研究生在读.E-mail:1158880238@qq.com
基金资助:
国家自然科学基金（81573725）；安徽省自然科学基金（1408085MH165，1508085MH163）

Inhibitory effects of chloroform extracts of Longdan xiegan decoction against hyphae formation of Candida albicans isolated from VVC

WANG Xia^1,2, WU Da-qiang^1,2, SHI Gao-xiang^1,2, DUAN Qiang-jun^1,2, SHAO Jing^1,2, WANG Tian-ming^1,2, WANG Chang-zhong^1,2

School of Integrated Traditional and Western Medicine, Anhui University of Chinese Medicine, Institute of Integrated Traditional and Western Medicine, Anhui Academy of Chinese Medicine, Hefei 230038

Received:2016-08-08 Online:2016-12-28 Published:2016-12-28

摘要/Abstract

摘要：

目的观察龙胆泻肝汤氯仿提取物（chloroform extracts of Longdan xiegan decoction，CELX）对白念珠菌菌丝形成的影响，探讨其可能的作用机制。方法采用微量稀释法测定龙胆泻肝汤不同溶剂提取物对15株VVC临床株的最低抑菌浓度（minimal inhibitory concentration，MIC）；采用固体培养基和半固体培养基观测菌落形态及菌丝侵袭能力；XTT还原法评价白念珠菌菌丝活性；平板法计数6 h内药物对白念珠菌CFU的影响；荧光显微镜下观察菌丝代谢活力；采用实时荧光定量PCR （qRT-PCR）检测其菌丝相关基因：ALS3、SSA1、SUN41、HWP1、UME6和ECE1的表达量。结果 CELX对15株VVC临床株MIC介于32~64 mg/L之间，效果优于总提物、石油醚提取物、乙酸乙酯提取物、正丁醇提取物和水提取物的MIC；256 mg/L的CELX能显著抑制菌丝的形成；256 mg/L的CELX在固体培养基中可抑制菌落褶皱，在半固体培养基中可抑制菌丝侵袭；qRT-PCR结果显示256 mg/L CELX可使菌丝相关基因ALS3、SSA1、SUN41、HWP1、UME6和ECE1分别下调81.6%、60.2%、69.8%、73.7%、49.5%、52.8%。结论龙胆泻肝汤氯仿提取物（CELX）可通过影响菌丝相关基因的表达来抑制菌丝的形成，从而降低白念珠菌对机体的侵袭力。

关键词: 龙胆泻肝汤, 白念珠菌, 外阴阴道念珠菌病, 菌丝

Abstract:

Objective To investigate the effects of chloroform extracts of Longdan xiegan decoction (CELX) against hyphae formation of Candida albicans clinical strains isolated from Vulvovaginal Candidiasis (VVC).Methods Microdilution method was used to determine the minimal inhibitory concentrations (MICs) of Longdan xiegan decoction extracts against C. albicans isolated from VVC. Solid agar and Semi-solid agar were utilized to evaluate colony morphology and invasive growth of C. albicans,respectively.XTT and Fluorescence microscope assay were applied to determine the metabolic activity of hyphae of C. albicans treated by CELX for 6 h.Plate assay counted CFU variation.Quantitative Real-time PCR (qRT PCR) was adopted to analyze the expression of hyphae-specific genes including ALS3,SSA1,SUN41,HWP1,UME6 and ECE1.Results The MICs of CELX against C. albicans strains were between 32 and 64 mg/L,which were better than MICs of total extract,petroleum ether extract,ethyl acetate extract,buty alcohol extract and water extract.CELX with concentration of 256 mg/L inhibited hyphae formation and influenced colony morphology significantly,with the wrinkle in solid agar and invasion in semi-solid agar being blocked.The expression of hyphae formation related genes,ALS3,SSA1,SUN41,HWP1,UME6 and ECE1 were down-regulated by 81.6%,60.2%,69.8%,73.7%,49.5%,52.8%.Conclusion Our results suggested that CELX could inhibit hyphae formation through repress hyphae-relative genes expression to reduce the invasive activity against patient.

Key words: Longdan xiegan decoction, Candida albicans, vulvovaginal candidasis (VVC), hyphae

中图分类号:

R379.4

王霞, 吴大强, 施高翔, 段强军, 邵菁, 汪天明, 汪长中. 龙胆泻肝汤氯仿提取物对白念珠菌VVC临床株菌丝抑制作用研究[J]. 中国真菌学杂志, 2016, 11(6): 341-347.

WANG Xia, WU Da-qiang, SHI Gao-xiang, DUAN Qiang-jun, SHAO Jing, WANG Tian-ming, WANG Chang-zhong. Inhibitory effects of chloroform extracts of Longdan xiegan decoction against hyphae formation of Candida albicans isolated from VVC[J]. Chinese Journal of Mycology, 2016, 11(6): 341-347.

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