去甲亚精胺对白色念珠菌生物膜形成的抑制作用

The inhibiting effect of norspermidine on Candida albicans biofilm formation

目的:研究去甲亚精胺对白色念珠菌生物膜形成的作用及可能的机制。方法:通过结晶紫染色、甲基四氮盐(XTT)检测、菌落形成单位(CFU)计数、扫描电镜(SEM)成像和qRT-PCR分别检测或观察去甲亚精胺对白色念珠菌生物膜生物量、代谢活性、菌落数、真菌形态和毒力相关基因表达的影响。结果:结晶紫染色结果显示不同浓度去甲亚精胺对白色念珠菌生物膜形成的抑制作用不同;XTT检测结果表明去甲亚精胺能显著抑制白色念珠菌生物膜的代谢活性;CFU计数结果显示去甲亚精胺能显著减少生物膜中白色念珠菌的菌落数;SEM结果显示去甲亚精胺能显著抑制生物膜形成,高浓度的去甲亚精胺能促进酵母相形成;q RT-PCR结果显示去甲亚精胺能显著抑制白色念珠菌毒力相关基因hwp、als3、csh1的表达。结论:去甲亚精胺能显著抑制白色念珠菌生物膜形成的作用并抑制毒力基因的表达,是一种潜在的抑制白色念珠菌生物膜的多胺类药物。

Objective: To study the effect of norspermidine on Candida albicans (C.albicans) biofilm formation and the underlying mechanisms. Methods: Crystal violet staining, XTT assay, colony-forming unit (CFU) count, scanning electron microscope (SEM) imaging and qRT-PCR was conducted to detect the effect of norspermidine on biofilm biomass, metabolic activity, colony number, fungal morphology and virulence-related gene expression of C.albicans biofilm. Results: Crystal violet staining results demonstrated that different concentrations of norspermidine had different inhibitory effects on C.albicans biofilm formation. XTT assay showed that norspermidine significantly restrained the metabolic activity of C.albicans biofilm. CFU counting results indicated that norspermidine severely reduced the number of C.albicans colonies in biofilm. SEM imaging displayed that norspermidine dramatically disturbed the formation of C.albicans biofilm and norspermidine in high concentration promoted the occurrence of yeast phase;qRT-PCR showed that norspermidine substantially interfered with the gene expression of ywp, hwp and als3, which were related to the virulence of C.albicans. Conclusion: Norspermidine could be used as a potential polyamine drug for anti-C.albicans biofilm as it could not only inhibit the formation of C.albicans biofilm but also interfere with the expression of its virulence-associated gene expression.