摘要
目的：探讨白头翁汤正丁醇提取物（butyl alcohol extract of Baitouweng decoction,BAEB）在碱性条件下对白念珠菌VVC（vulvovaginal candidiasis）临床分离株酵母-菌丝形态转化的影响. 方法：采用肉汤稀释法测定白头翁汤不同溶剂萃取物对VVC临床株的最低抑菌浓度（minimal inhibitory concentration,MIC）;倒置显微镜观察不同pH环境下VVC临床株形态;采用XTT还原法评价在pH 8.0时菌丝活性;扫描电镜观察白念珠菌菌丝形态;荧光显微镜观察菌体代谢活力;固体培养基上观测菌落形态;半固体培养基上观察菌丝侵袭能力;实时荧光定量PCR（qRT-PCR）检测菌丝特异性基因ALS3,CSH1,SUN41,HWP1和CaPDE2的表达量. 结果：白头翁汤正丁醇提取物（BAEB）对VVC临床株的MIC为64~128 mgL-1,低于总提取物、石油醚提取物、乙酸乙酯提取物和水提取物的MIC;pH 8.0时VVC临床株的菌丝形成能力最强;扫描电镜结果显示512, 1 024 mgL-1的BAEB明显抑制VVC临床株的形态转化,荧光显微镜结果显示1 024 mgL-1的BAEB可以降低VVC临床株的代谢活性;512,1 024 mgL-1BAEB浓度的固体菌落未出现褶皱,1 024 mgL-1BAEB浓度的半固体培养基内部未见菌丝侵袭;qRT-PCR检测显示1 024 mgL-1时,ALS3,CSH1,SUN41,HWP1分别下调4.26,3.2,2.74,5.12倍,CaPDE2上调2.38倍. 结论：BAEB在碱性pH条件下对白念珠菌VVC临床株的形态转化具有抑制作用.
Objective： To investigate the effects of butyl alcohol extract of Baitouweng decoction （BAEB） on yeast-to-hyphae transition of Candida albicans isolates from vulvovaginal candidiasis （VVC） in alkaline pH. Method： Serial 2-fold dilution assay was used to determine the minimal inhibitory concentrations（MICs） of Baitouweng decoction extracts against C. albicans isolates from VVC, XTT assay was applied to determine the metabolic activity of C. albicans hypha treated by BAEB for 6 h. The morphological change of C. albicans treated by BAEB was inspected at different pH by inverted microscope, fluorescence microscope, scanning electron microscopy（SEM）. Solid agar plate and semi-solid agar were utilized to evaluate colony morphology and invasive growth of C. albicans, respectively. Quantitative Real-time PCR （qRT-PCR） was adopted to observe the expressions of hyphae-specific genes including HWP1, ALS3, CSH1, SUN41 and CaPDE2. Result： The MIC of BAEB against C. albicans is less than that of other extracts; hyphae grow best at pH 8.0; 512 mgL-1 and 1 024 mgL-1 BAEB could inhibit formation of hyphae and influence colony morphology. When treated by 512 mgL-1 and 1 024 mgL-1 BAEB, the colonies became smooth; while by 0 and 256 mgL-1 BAEB, the colonies became wrinkled. In semi-solid agar, the length of hyphae decreased steadily as the concentration of BAEB lowered. The expression of HWP1,ALS3,CSH1,SUN41 were downregulated by 5.12, 4.26, 3.2 and 2.74 folds, and CaPDE2 was upregulated by 2.38 fold. Conclusion： BAEB could inhibit yeast-to-hyphae transition of C. albicans isolates from VVC in alkaline pH.keywords：butyl alcohol extract of Baitouweng decoction（BAEB）
出处
《中国中药杂志》
CAS
CSCD
北大核心
2015年第4期710-715,共6页
China Journal of Chinese Materia Medica
基金
安徽省自然科学基金项目(1408085MH165)
安徽中医药大学校级科研项目(2014zr026)
