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目的观察云南重楼正丁醇提取物(BEPP)对体外白念珠菌生物膜形成的影响。方法微量稀释法测定BEPP最低抑菌浓度(MIC),XTT法测定生物膜抑制80%浓度(SMIC80),平板法绘制时间-杀菌曲线,扫描电镜(SEM)观察生物膜形态结构,激光共聚焦显微镜(CLSM)观测生物膜厚度,实时荧光定量PCR(q RT-PCR)法检测生物膜形成相关基因HWP1、MP65、SUN41表达量的变化。结果 BEPP对白念珠菌的MIC为32~128μg/m L,对白念珠菌生物膜的SMIC80为128~512μg/m L,时间-杀菌曲线显示BEPP具有良好的杀菌作用,SEM观察到BEPP有效抑制白念珠菌生物膜形成,CLSM显示BEPP可以降低生物膜厚度,q RT-PCR检测显示在BEPP作用下,HWP1、MP65、SUN41基因表达量均显著下调。结论 BEPP可以有效抑制体外白念珠菌生物膜的形成。
Objective To observe the effect of BEPP on the formation of C. albicans biofilm in vitro. Methods The minimum inhibitory concentration (MIC) of BEPP was determined by microdilution method. The biofilm inhibitory concentration (SMIC80) was determined by XTT method. The time-bactericidal curve was drawn by plate method. The morphology of biofilm was observed by scanning electron microscopy (SEM) (CLSM) biofilm thickness, and the changes of biofilm formation-related genes HWP1, MP65 and SUN41 were detected by qRT-PCR. Results The MIC of BEPP to Candida albicans was 32 ~ 128μg / mL and the SMIC80 of Candida albicans biofilm was 128 ~ 512μg / mL. The time-kill curve showed that BEPP had a good bactericidal activity. The SEM observation showed that BEPP effectively inhibited the growth of Candida albicans CLSM showed that BEPP could reduce the biofilm thickness. QRT-PCR analysis showed that the expression of HWP1, MP65 and SUN41 were down-regulated significantly under the action of BEPP. Conclusion BEPP can effectively inhibit the formation of C. albicans biofilm in vitro.