黄根醇提取物体外抗白色念珠菌生物膜作用的实验研究

摘要

目的：观察黄根醇提取物体外对白色念珠菌生物膜及相关基因的影响。方法 M27-A2测定黄根醇提取物对白色念珠菌的最小抑菌浓度（MIC）；XTT法评价黄根醇提取物对白色念珠菌生物膜形成的影响；实时荧光定量RT-PCR（qRT-PCR）检测黄根醇提取物作用后，ALS基因的表达情况。结果黄根醇提取物对白色念珠菌的MIC为8μg/mL；随着浓度的增加，黄根醇提取物对白色念珠菌生物膜的抑制作用增强，呈正相关性；16μg/mL黄根醇提取物对不同生长阶段的白色念珠菌生物膜均有抑制作用，抑制率随生物膜成熟逐渐降低；qRT-PCR结果显示，药物作用后ALS2、ALS3基因表达降低（7.87±0.27比5.15±0.34；6.24±0.51比2.13±0.23，P<0.05），ALS1基因表达无明显变化（6.11±0.14比5.95±0.31，P>0.05）。结论黄根醇提取物对体外白色念珠菌生物膜有较明显的抑制作用，可能通过抑制ALS2、ALS3基因的表达而实现。%Objective To study the effect of alcohol extract from Prismatomeris tetrandra on candida albicans(C. albicans) biofilms and the expression of related genes in vitro. Methods M27-A2 was used to determine the min-imum inhibitory concentration (MIC) of alcohol extract from Prismatomeris tetrandra against C. albicans. XTT assay was performed to determine the effects of alcohol extract from Prismatomeris tetrandraon C.albicans biofilm forma-tion. The real-time fluorescent quantitative RT-PCR(qRT-PCR) was used to determine the difference of ALS gene expression between before and after alcohol extract from Prismatomeris tetrandra induction group. Results MIC of alcohol extract from Prismatomeris tetrandra against C. albicans was 8μg/mL. With increasing concentration, the in-hibitory effect of alcohol extract from Prismatomeris tetrandra on C. albicans biofilms enhanced. Alcohol extract from Prismatomeris tetrandra at concentration of 16μg/mL showed distinct inhibitive effect on adhesion to C.albicans cultured for 4h, 8h, 12h, 24h and 48h, and the effect weakened as the time of culture increased. qRT-PCR re-sults showed that alcohol extract from Prismatomeris tetrandra resulted in a striking down-regulation of ALS2 and ALS3 (ALS2:7.87 ±0.27 vs 5.15 ±0.34; ALS:6.24 ±0.51 vs 2.13 ±0.23; all P<0.05), while the expression of ALS1 showed no changes (6.11 ±0.14 vs 5.95 ±0.31, P>0.05). Conclusion Alcohol extract from Prismatomeris tetrandra has potent activity against C.albicans biofilms in vitro. The inhibition of alcohol extract from Prismatomeris tetrandra on biofilms formation may depend on suppressing the expression of ALS2 and ALS3.