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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 the effect of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) on the adhesion of Candida glabrata. Methods: The minimum inhibitory concentration (MIC) of extract of Huanglian Jiedu decoction against Candida glabrata was determined by double dilution method. The effect of EAHD on the adhesion of Candida glabrata was evaluated by XTT reduction method. The morphological changes of Candida glabrata were observed by microscope, scanning electron microscope and fluorescence microscope. The expression of adhesion related genes (EPA1, EPA6 and EPA7) were detected by semi-quantitative and real-time fluorescence quantitative PCR. Results: The MIC of EAHD to Candida glabrata was 320 mg · L-1, the MIC for positive control fluconazole against Candida glabrata was 1 mg · L-1; and the adherence of EAHD to Candida glabrata for 2 and 4 h The effect was obvious at 4 h, which was better than 2 h. The results of PCR showed that EAHD significantly inhibited the expression of EPA1, EPA6 and EPA7. Conclusion: EAHD may inhibit its early adhesion by inhibiting the expression of some adhesion genes of Candida.