食品科学 ›› 2023, Vol. 44 ›› Issue (19): 18-26.doi: 10.7506/spkx1002-6630-20220512-148

• 基础研究 • 上一篇    

二氢槲皮素对大肠杆菌的抑菌作用机理

蔡瑾,闫然,王梦亮,王琪   

  1. (1.山西大学应用化学研究所,山西 太原 030006;2.山西大学中医药现代研究中心,山西 太原 030006;3.山西大学 化学生物学与分子工程教育部重点实验室,山西 太原 030006;4.山西大学生命科学学院,山西 太原 030006)
  • 发布日期:2023-11-07
  • 基金资助:
    山西省基础研究计划自然科学研究面上项目(20210302123451)

Antimicrobial Mechanism of Dihydroquercetin against Escherichia coli

CAI Jin, YAN Ran, WANG Mengliang, WANG Qi   

  1. (1. Institute of Applied Chemistry, Shanxi University, Taiyuan 030006, China; 2. Modern Research Center for Traditional Chinese Medicine, Shanxi University, Taiyuan 030006, China; 3. The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, Taiyuan 030006, China; 4. School of Life Science, Shanxi University, Taiyuan 030006, China)
  • Published:2023-11-07

摘要: 食源性病菌给食品安全带来重大的挑战。二氢槲皮素(dihydroquercetin,DHQ)可作为食品防腐剂,但目前鲜见其抑菌机理的相关报道。本实验发现DHQ对6 种食源性致病菌,即金黄色葡萄球菌、枯草芽孢杆菌、大肠杆菌、普通变形杆菌、产气肠杆菌和热带假丝酵母菌均有显著抑制作用(P<0.05),且对大肠杆菌的抑制活性最佳,故以大肠杆菌为指示菌,研究DHQ的抑菌机理。利用扫描电子显微镜和透射电子显微镜观察,发现在DHQ作用下的大肠杆菌严重变形,表现出黏连、折叠、质壁分离、出现空泡结构等现象。荧光探针被用来测定膜内活性氧(reactive oxygen species,ROS)水平,结果显示DHQ能显著增加细胞内的ROS水平(P<0.05)。利用Annexin V-FITC/PI试剂盒检测细胞膜的通透性和完整性,发现随着DHQ质量浓度增加,细胞膜通透性显著增加(P<0.05),细胞膜的完整性遭到破坏。使用罗丹明123检测细胞膜电位,结果表明DHQ会引起大肠杆菌细胞膜去极化,造成膜电位显著降低(P<0.05)。细胞膜内Ca2+检测结果表明,DHQ会导致胞内Ca2+外漏,影响菌体的正常生长代谢和功能活动。采用碘化丙啶/核糖核酸酶(propidium iodide/ribonuclease,PI/RNase)染色缓冲液检测细胞周期,结果表明DHQ干扰了大肠杆菌正常的细胞周期。研究结果揭示了DHQ主要作用于细胞膜,影响正常传代,从而达到抑制大肠杆菌的效果。

关键词: 二氢槲皮素;抑菌活性;大肠杆菌;抑菌机理

Abstract: Foodborne pathogens pose a major challenge to food safety. Dihydroquercetin (DHQ) can be used as a food preservative; however, its antimicrobial mechanism is still unclear. This study found that DHQ had significant antimicrobial activity against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Proteus vulgaris, Enterobacter aerogenes and Candida tropicalis (P < 0.05), the effect being most pronounced against E. coli. The antimicrobial mechanism against E. coli was investigated. Observation by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) revealed that the DHQ-treated cells became distorted and exhibited phenomena such as adhesion, folding, plasmolysis, and the occurrence of vacuoles. DHQ resulted in a significant increase in the level of reactive oxygen species (ROS) inside the cell membrane as determined using a fluorescence probe. The determination of Annexin V-FITC/P kit showed that cell membrane permeability significantly increased with increasing DHQ concentration (P < 0.05) and cell membrane integrity was damaged. Cell membrane potential measurement by Rhodamine 123 staining revealed that DHQ caused cell membrane depolarization, leading to a significant reduction in membrane potential (P < 0.05). Moreover, DHQ caused intracellular Ca2+ leakage, which perturbed the growth, metabolism and functional activity of E. coli. DHQ interfered with the cell cycle of E. coli as determined using propidium iodide/ribonuclease (PI/RNase) staining buffer. These findings revealed that DHQ can inhibit E. coli growth mainly by acting on the cell membrane and consequently affecting the normal passage of cells.

Key words: dihydroquercetin; antimicrobial activity; Escherichia coli; antimicrobial mechanism

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