桑黄正丁醇提取物对白念珠菌生物膜形成的抑制作用
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安徽省科技厅国家技术创新工程试点项目(20140741);安徽省高校自然科学研究重点项目(KJ2017A287)


Inhibitory Effect of the Butyl Alcohol Extract of Phellinus igniarius on Candida albicans Biofilm Formation
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    目的 探讨桑黄正丁醇提取物(butyl alcohol extract of Phellinus igniarius decoction,BAEP)体外对白念珠菌(Candida albicans)标准株SC5314生物膜形成的影响。 方法 采用二倍稀释法测定BAEP对白念珠菌的最低抑菌浓度(minimal inhibitory concentration,MIC)和抑制50%生物膜形成的最低浓度(sessile minimal inhibitory concentration,SMIC50);采用XTT还原法测定BAEP对白念珠菌生物膜代谢的影响;固体培养基上观察BAEP对白念珠菌菌落形态的影响;倒置显微镜与荧光显微镜下分别观察BAEP对白念珠菌生物膜形态与活性的影响;扫描电镜下观察BAEP对白念珠菌生物膜形态结构的影响;实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)检测生物膜相关基因ALS1和HWP1的转录水平变化。结果 BAEP对白念珠菌生物膜的SMIC50为1 024 μg/mL;菌落形态实验观察结果显示,1 024 μg/mL BAEP可影响白念珠菌菌落;倒置显微镜、荧光显微镜下显示1 024 μg/mL BAEP可抑制白念珠菌生物膜的形态和活性;扫描电镜下显示1 024 μg/mL BAEP对白念珠菌生物膜有明显的抑制作用;qRT-PCR检测结果显示BAEP可明显下调HWP1的表达水平和上调ALS1基因的表达水平。结论 BAEP对白念珠菌SC5314生物膜的形成有一定的抑制作用。

    Abstract:

    Objective To investigate the inhibitory effect of the butyl alcohol extract of Phellinus igniarius decoction (BAEP) on the biofilm formation of Candida albicans standard strain SC5314 in vitro. Methods The double-dilution method was used to determine the minimal inhibitory concentration (MIC) and sessile minimal inhibitory concentration (SMIC50) of BAEP for C. albicans. XTT assay was used to analyze the influence of BAEP on the metabolic activity of C. albicans SC5314 biofilm. The solid medium was used to observe the effect of BAEP on the colonial morphology of C. albicans. An inverted microscope and a fluorescence microscope were used to investigate the effect of BAEP on the morphology and activity of C. albicans SC5314 biofilm, and a scanning electron microscope was used to observe the effect of BAEP on the morphological structure of C. albicans biofilm. Quantitative real-time PCR (qRT-PCR) was used to observe the changes in the transcriptional levels of biofilm-associated genes ALS1 and HWP1. Results The SMIC50 of BAEP was 1 024 μg/mL for C. albicans biofilm. The results of colonial morphology showed that BAEP 1 024 μg/mL affected the colony of C. albicans. BAEP 1 024 μg/mL inhibited the morphology and activity of C. albicans biofilm under the inverted microscope and the fluorescence microscope. The scanning electron microscope showed that BAEP 1 024 μg/mL had a marked inhibitory effect on C. albicans biofilm. The results of qRT-PCR showed that BAEP significantly reduced the expression of HWP1 and increased the expression of ALS1. Conclusion BAEP exerts a certain inhibitory effect on the biofilm formation of C. albicans SC5314.

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汪天明,邸 磊,施高翔,刘慧敏,汪长中,李 宁.桑黄正丁醇提取物对白念珠菌生物膜形成的抑制作用[J].安徽中医药大学学报,2019,38(6):52-56

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  • 在线发布日期: 2019-11-25