干酪乳酸杆菌代谢物对白假丝酵母菌生物被膜的体外抑菌效果及分子机制

doi:10.6040/j.issn.1671-7554.0.2015.373

山东大学学报（医学版） ›› 2017, Vol. 55 ›› Issue (2): 74-78.doi: 10.6040/j.issn.1671-7554.0.2015.373

干酪乳酸杆菌代谢物对白假丝酵母菌生物被膜的体外抑菌效果及分子机制

冯凡,胡晓燕,吴伟芳,孙筱林,孙允东,曹倩,肖颖,闫世坤,史培堃,曾贝妮,周亚滨

山东大学医学院微生物学教研室, 山东济南 250012

收稿日期:2015-04-13 出版日期:2017-02-10 发布日期:2017-02-10
通讯作者: 周亚滨. E-mail:zyb@sdu.edu.cn E-mail:zyb@sdu.edu.cn
基金资助:
山东省自然科学基金(ZR2009CM073)

Antifungal effects and relevant mechanisms of metabolites of Lactobacillus casei against Candida albicans biofilms in vitro

FENG Fan, HU Xiaoyan, WU Weifang, SUN Xiaolin, SUN Yundong, CAO Qian, XIAO Ying, YAN Shikun, SHI Peikun, ZENG Beini, ZHOU Yabin

Department of Medical Microbiology, Medical School of Shandong University, Jinan 250012, Shandong, China

Received:2015-04-13 Online:2017-02-10 Published:2017-02-10

摘要/Abstract

摘要： 目的研究干酪乳酸杆菌代谢物对体外白假丝酵母菌生物被膜的抑菌作用及其机制。方法通过萃取、蒸馏的方式浓缩干酪乳酸杆菌代谢物。采用甲基四氮盐(XTT)减低法测定干酪乳酸杆菌代谢物对白假丝酵母菌YEM30游离态及生物被膜态的最小抑菌浓度(MIC₈₀),定量检测干酪乳酸杆菌代谢物对白假丝酵母菌体外代谢活动的影响,观察白假丝酵母菌形态学的转变,并通过双向凝胶电泳(2-DE)对比干酪乳酸杆菌代谢物处理前后白假丝酵母菌蛋白的表达差异。结果干酪乳酸杆菌代谢物对白假丝酵母菌的游离态、生物被膜态的最小抑菌稀释倍数分别为1∶256和1∶128。稀释倍数为1∶256的干酪乳酸杆菌代谢物可以明显干扰白假丝酵母菌酵母相向菌丝相的转化。比较双向电泳图谱上获得6个显著蛋白斑点,对其中的4个进行鉴定,结果显示在干酪乳酸杆菌代谢物的刺激下,烯醇酶、苹果酸脱氢酶、泛素连接酶表达升高,己糖激酶表达下降。结论干酪乳酸杆菌代谢物对体外白假丝酵母菌生物被膜具有抑制作用。

关键词: 抑菌作用, 干酪乳酸杆菌代谢物, 生物被膜, 双向电泳, 白假丝酵母菌

Abstract: Objective To detect the antifungal effects and relevant mechanisms of metabolites of Lactobacillus casei against Candida albicans biofilms in vitro. Methods Metabolites of Lactobacillus casei were obtained by extraction and distillation. MIC₈₀ of metabolites of Lactobacillus casei against planktonic cells and biofilms were determined with the tetrazolium salt(XTT)reduction assay. The metabolic activity of Candida albicans biofilms against metabolites of Lactobacillus casei was assessed quantitatively. Yeast-to-hypha morphological transition was examined qualitatively with inverted microscope. Two-dimensional gel electrophoresis(2-DE)was used to search differentially expressed protein spots against metabolites of Lactobacillus casei-treated Candida albicans. Results MIC₈₀ of metabolites of Lactobacillus casei against Candida albicans planktonic cells and biofilms were 1∶256 and 1∶128, respectively. When the dilution ratio was 1∶256, metabolites of Lactobacillus casei blocked the yeast-to-hypha transition. Compared with six significant protein spots achieved from the 2-DE maps, four of them were indentified. The expressions of enolase, ubiquitin ligase and malate dehydrogenase were increased, while the expression of hexokinase was decreased under the stimulation of metabolites of Lactobacillus casei. Conclusion Metabolites of Lactobacillus casei can inhibit Candida albicans biofilms in vitro.

Key words: Candida albicans, Metabolite of Lactobacillus casei, Antifungal effects, Biofilm, Two-dimensional gel electrophoresis

中图分类号:

R379.4

冯凡,胡晓燕,吴伟芳,孙筱林,孙允东,曹倩,肖颖,闫世坤,史培堃,曾贝妮,周亚滨. 干酪乳酸杆菌代谢物对白假丝酵母菌生物被膜的体外抑菌效果及分子机制[J]. 山东大学学报（医学版）, 2017, 55(2): 74-78.

FENG Fan, HU Xiaoyan, WU Weifang, SUN Xiaolin, SUN Yundong, CAO Qian, XIAO Ying, YAN Shikun, SHI Peikun, ZENG Beini, ZHOU Yabin. Antifungal effects and relevant mechanisms of metabolites of Lactobacillus casei against Candida albicans biofilms in vitro[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2017, 55(2): 74-78.

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干酪乳酸杆菌代谢物对白假丝酵母菌生物被膜的体外抑菌效果及分子机制

Antifungal effects and relevant mechanisms of metabolites of Lactobacillus casei against Candida albicans biofilms in vitro

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