黄连解毒汤乙酸乙酯提取物对光滑念珠菌黏附作用的影响

Anti-attachment effect of ethyl acetate extract of Huanglian Jiedu decoction on Candida glabrata

目的:探讨黄连解毒汤乙酸乙酯提取物(ethyl acetate extract of Huanglian Jiedu decoction,EAHD)对光滑念珠菌Candida glabrata黏附作用的影响。方法:采用二倍稀释法测定黄连解毒汤乙酸乙酯提取物对光滑念珠菌的最低抑菌浓度(minimum inhibitory concentration,MIC);采用XTT还原法评价EAHD对光滑念珠菌黏附作用的影响;利用倒置显微镜、扫描电子显微镜与荧光显微镜观察EAHD对光滑念珠菌早期黏附形态学影响;采用半定量与实时荧光定量PCR检测其黏附相关基因EPA1,EPA6和EPA7的表达量。结果:EAHD对光滑念珠菌的MIC为320 mg·L-1,阳性对照药氟康唑对光滑念珠菌的MIC为1 mg·L-1;EAHD对光滑念珠菌2,4 h的黏附的干预作用较明显,其中对4 h效果优于2 h;PCR结果显示EAHD可显著抑制EPA1,EPA6和EPA7的表达。结论:EAHD可能通过抑制光滑念珠菌部分黏附基因的表达而抑制其早期黏附。

Objective： To investigate anti-attachment effect of ethyl acetate extract of Huanglian Jiedu decoction（EAHD） on Candida glabrata. Method： Serial 2-fold dilution assay was used to determine the minimum inhibitory concentrations MICs of EAHD to C. glabrata. XTF assay was used to evaluate the effect of EAHD against adhesion of C. glabrata. Inverted microscope, scanning electron microscope （SEM） and fluorescein diacetate （FDA） staining were applied to observe the morphological changes of C. glabrata in adhesion. PCR was adopted to inspect the expression of attachment-related genes such as EPA1, EPA6 and EPA7. Result： The MIC of EAHD and fluconazole to C. glabrata were 320 mg·L-1 and 1 mg·L-1 respectively. The total cells including budding ceils decreased in a dose-dependent manner following EAHD treatment. The expressions of EPAI, EPA6 and EPA7 were downregulated dra- matically after EAHD treatment. Conclusion： EAHD could effectively inhibit adherence of C. glabrata.